Hybrid nanostructure designs facilitated by M13 virus for lithium ion battery and lithium air battery electrodes By Dahyun Oh B.S. Materials Science and Engineering, Seoul National University, Korea, 2008 SUBMITTED TO THE DEPARTMENT OF MATERIALS SCIENCE AND ENGINEERING IN PARTIAL FULFILLMENT OF THE REQUIREMENT FOR THE DEGREE OF DOCTOR OF PHILOSOPHY MASSACHUSETTS INSTITUTE OF TECHNOLOGY JANUARY 2014 0 2014 Massachusetts Institute of Technology. All rights reserved. -MASSAHUStTS TE OF TECHNOLOGY MAY 14 2014 LIBRARIES Signature of Author: Department of M rials Science and Engineering January 1 7 th 2014 Certified by: Angela M. Belcher Materials Science and Engineering and Biological Engineering Thesis Supervisor Accepted by: Chair, Departmental C on Hybrid nanostructure designs facilitated by M 13 virus for lithium ion battery and lithium air battery electrodes By Dahyun Oh B.S. Materials Science and Engineering, Seoul National University, Korea, 2008 Submitted to the Department of Materials Science and Engineering on Jan 17 th, 2014 in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in Materials Science and Engineering at the Massachusetts Institute of Technology ABSTRACT The development of technology and population growth will demand 56 percent increase of the energy consumption in 30 years. An efficient energy storage system will be necessary to meet these increased needs to deliver and store the energy. After the first release of commercial Li ion batteries in 1991, they were widely adapted to various applications from small portable devices to electric vehicles. However, the current Li ion battery can only store -250 Wh/kgcelj of gravimetric energy, a far limited energy storage capability especially to replace gasoline in powering vehicles. This limitation originated either from the incomplete utilization of active materials or their low theoretical energy density. Therefore, a rational design of electrodes as well as the new battery chemistry needs to be investigated to further develop the current energy storage system. In this thesis, high theoretical energy density batteries are investigated. First, the power performance of conversion reaction cathode materials, bismuth oxyfluorides, was improved. By rationally designing genetic sequences of the M13 virus, graphene sheets were homogeneously distributed throughout bismuth oxyfluorides cathodes as conducting paths. Second, large surface area cathodes were developed with virus-templated manganese oxide nanowires. These electrodes were applied to Li-0 2 battery systems to achieve large capacities and a long cycle life. Furthermore, the chemical composition of virus-templated inorganic nanowires was easily tuned to study the catalytic behavior of transition metal oxides in Li-0 2 batteries. These bio-directed methods to develop high performance battery electrodes, in conclusion, suggest an eco-friendly and cost effective way to manufacture energy storage devices. The design strategy established in this thesis could be applied not only to batteries but also to electronic devices requiring sophisticated nanoscale controls. Thesis advisor: Angela M. Belcher Title: W.M. Keck Professor of Energy 1 ACKNOWLEDGEMENT It was a great journey to complete Ph.D. program at MIT, full of excitement, curiosity and endurance. For five years of study, Prof. Angela M. Belcher, who is my teacher and advisor, has led me to learn science and technology but also life. I would like to send my sincere thanks to Angie for her strong support, encouragement and passion. In addition to my advisor, it was a great pleasure to meet many insightful scholars at MIT including my thesis committee members, Prof. Gerbrand Ceder, Prof. Paula T. Hammond and my course work teacher, Prof. Michael J. Demkowicz, Prof. Samuel M. Allen. I appreciate their efforts and time to guide me to successfully finish my thesis and course work. As a graduate student researcher, I had many great opportunities to communicate and discuss about the research with Prof. Yang Shao-Horn and Prof. Jing Kong. Their feedback and guidance has directed me to find excitements in research. In addition to leading investigators, I would like to thank to my colleagues, collaborators and friends at MIT. I spent most of my school life at CMSE and ISN, with the help from Dr. Yong Zhang, Dr. Scott Speakman and Mr. William F. DiNatale. My lab mates, Prof. Yun Jung Lee, Dr. Hyunjung Yi, Prof. Debadyuti Ghosh, Dr. Gaelen Hess and my neighbor Mr. John Patrick Casey have kindly shared all of my joy, sadness and worries for my graduate life. Furthermore, I am thankful to Dr. Jifa Qi, Prof. Yi-Chun Lu, Dr. Nimrod Heldman, Prof. Seung Woo Lee and Prof. Byungwoo Kang for their scientific feedback and kind answers to my endless questions. My Korean classmates, Eun Seon, YJ, Yong-cheol and international classmates, Billy, Jo, Kevin, Marco, Sophie, thank you all to spend the most exciting MIT course 3 graduate life together. I am thankful to my friend at MIT, Jouha, Sungmin, Chris, Yunna, Su, Dong Sook, Eun hee to mentally support my graduate life. Finally, I appreciate the financial support from Kwanjeong Educational Foundation and Prof. Anne Mayes. Most of all, I would like to send my sincere thank to my family, Rosia, George, Ann, Dongho for their kind greetings and welcoming during my vacation in US. I appreciate the support from my lovely aunts in Korea as well. I am sending my greatest love to my only one sister, Jihyun for sharing all pains, love and encouragements. I am also sending my deepest sorry, love and thank to my parents in Korea for their firmness, health, support and love. In final, I am grateful to God, who is the start and the end, to lead me to his love. 2 BIOGRAPHICAL NOTE EDUCATION 2008.09 ~ present Ph. D. at Department of Materials Science and Engineering Massachusetts Institute of Technology (MIT), Massachusetts, USA Advisor: Angela M. Belcher Ph. D. thesis title: Hybrid nanostructure designs facilitated by M 13 virus for lithium ion battery and lithium air battery electrodes 2004.03 ~ 2008.01 B.S. at Department of Materials Science and Engineering Seoul National University, Seoul, Korea Graduated with honors (Summa cum laude) AWARDS AND SCHOLARSHIPS 2013 2011 2008-2013 2008 2007 2006 2005 2004 Graduate Student Award, Silver, Fall MRS Meeting, Boston, USA Outstanding Poster Award, Fall MRS Meeting, Boston, USA Kwanjeong Educational Foundation graduate student fellowship, Seoul, Korea Mayes fellowship, MIT, USA Korea Semiconductor Industry Association fellowship, Seoul, Korea Korea Federation of Textile Industries fellowship, Seoul, Korea Honors and fellowship for top student, SNU, Seoul, Korea Korea Science and Engineering Foundation fellowship, Seoul, Korea PUBLICATION Journal Publications 1. Dahyun Oh, Jifa Qit, Geran Zhang, Thomas Carney, Yang Shao-Horn* and Angela M. Belcher* (*Equal contribution) Variations of oxygen reduction/oxygen evolution reactions in Li-0 2 batteries with biotemplated multi-component (Mn,Co) transition metal oxide nanowires, In preparation, 2014 2. Dahyun Oh, Jifa Qit, Yi-Chun Lu, Yong Zhang, Yang Shao-Horn* and Angela M. Belcher* ( Equal contribution) Biologically enhanced cathode design for improved capacity and cycle life for Li-oxygen batteries, Nature communications, 2013, 4, 2756 DOI: 10.1038/ncomms3756 3. Dahvun Oh, Xiangnan Dang, Hyunjung Yi, Mark A. Allen, Kang Xu, Yun Jung Lee* and Angela M. Belcher* Graphene sheets stabilized on genetically engineered M 13 viral templates as Conducting Frameworks for Hybrid Energy-Storage Materials, Small, 2012, 8 (7), pp 1006-1011 DOI: 10.1002/smll.20 1102036 4. Yun Jung Leet, Youjin Leet, Dahyun Oh, Tiffany Chen, Gerbrand Ceder* and Angela M. Belcher* (tEqual contribution) Biologically Activated Noble Metal Alloys at the Nanoscale: For Lithium Ion Battery Anodes Nano Letters, 2010, 10 (7), pp 2433-2440 DOI: 10.1021/n11005993 3 Conference Proceedings 1. Kang Xu*, Dahvun Oh, Hyunjung Yi, Jifa Qi, Alice Xu, James Snyder and Angela M. Belcher, ECS transaction, 2012, 41 (41), pp 55-64 DOI: 10. 1149/1.4717963 Genetically Programming Interfaces between Active Materials, Conductive Pathway and Current Collector in Li Ion Batteries PATENT 1. Dahvun Oh, Jifa Qi, Yang Shao-Horn and Angela M. Belcher M 13 bacteriophage templated nanocomposites of transition metal oxides and metal nanoparticles for high performance lithium air batteries, US patent filed. 4 TABLE OF CONTENTS Abstract ............................................................................................................................................ Acknow ledgem ent ........................................................................................................................... Biographical note ............................................................................................................................. L ist of Tables ................................................................................................................................... Lisgt of Figures .................................................................................................................................. I 2 3 6 6 Chapter 1. Introduction .................................................................................................................... 1. 1 Background ........................................................................................................................... 1.2 Scopeofw ork ..................................................................................................................... 1.3 References ........................................................................................................................... 8 9 12 13 Chapter 2. Integrating graphene based conducting frameworks into Lithium ion battery cathodes (Bism uth oxyfluorides, BiOo. 5F 2) by genetic engineering of M 13 virus ................................... 15 2.1 Introduction ......................................................................................................................... 16 2.2 Experim ental ....................................................................................................................... 18 2.3 Results and discussion ..................................................................................................... 23 2.4 Conclusion ........................................................................................................................... 30 2.5 Figures and tables ................................................................................................................ 31 2.6 References ............................................................................................................................ 43 Chapter 3. Li-0 2 battery hybrid nanocatalyst formed with M 13 virus templated Manganese oxides nanow ires and noble m etal nanoparticles .................................................................................. 46 3.1 Introduction ......................................................................................................................... 47 3.2 Experim ental ....................................................................................................................... 49 3.3 Results and discussion ..................................................................................................... 54 3.4 Conclusion ........................................................................................................................... 64 3.5 Figures and tables ................................................................................................................ 65 3.6 References ............................................................................................................................ 84 Chapter 4. Investigation of catalytic behavior of M13 virus templated MnCo30 4 (x = 0, 1,2) nanow ires for Li-0 2 battery cathodes ....................................................................................... 89 4.1 Introduction ......................................................................................................................... 90 4.2 Experim ental ........................................................................................................................ 92 4.3 Results and discussion ..................................................................................................... 96 4.4 Conclusion ......................................................................................................................... 100 4.5 Figures and tables .............................................................................................................. 101 4.6 References .......................................................................................................................... 106 Chapter 5. Ongoing work: Probing graphene defects with genetic modifications of M13 virus ...................................................................................................................................................... 4.1 4.2 4.3 4.4 4.5 Introduction ....................................................................................................................... Experim ental ..................................................................................................................... Results and discussion ....................................................................................................... Figures and tables .............................................................................................................. References ......................................................................................................................... 5 10 8 109 110 II1 112 115 List of tables Table 2.1. XRD peaks table for synthesized bismuth oxyfluoride from 20-70 degree (2 Theta) with Rietveld refinement results for compound ....................................................................... 42 Table 2.2. Atomic concentration (%) of bismuth oxyfluoride by XPS analysis ....................... 42 Table 3.1. The Gas Chromatography (GC) data of Pd (3 wt %)/bio MO nanowires electrodes after the first galvanostatic discharge/charge at 0.4 A g-'c with 0.1 M LiCIO 4 in DME ................... 83 Table 3.2. The discharge/charge capacity (mAh g'c) for the first and second cycle of different electrodes galvanostatically tested at 0.4 A g-1, with 0.1 M LiCIO 4 in DME under I atm of 02 ....................................................................................................................................................... 83 List of figures Figure 2. 1. Characterizations of graphene................................................................................. 31 Figure 2.2. Determination of virus mass by Thermogravimetric analysis (TGA) .................... 32 Figure 2.3. The graphene/M 13 virus complex with the enhanced colloidal stability for the hybrid graphene/nanoparticle nanocom posites ..................................................................................... 33 Figure 2.4. Characterizations of the bismuth oxyfluoride nucleated on the graphene/virus complex ....................................................................................................................................................... 35 Figure 2.5. Characterizations of bismuth oxyfluoride with an electrochemical method and X RD ........................................................................................................................................................ 36 Figure 2.6. The power performance of the graphene/bismuth oxyfluoride nanocomposites with the genetically engineered M 13 virus ....................................................................................... 37 Figure 2.7. Electrochemical performance and characterization of bismuth oxyfluoride/graphene nanocomposites and bismuth oxyfluoride on FC#2 .................................................................. 39 Figure 2.8. XRD patterns of control samples ........................................................................... 40 Figure 2.9. The first five cycles galvanostatic data of bismuth oxyfluoride/graphene (2.4 wt%) nanocomposites (BOF-GP) with FC# 2 and control samples ................................................... 41 Figure 3.1. Schematic of a nanocomposite structure................................................................. 65 Figure 3.2. Electron microscope images of bio MO nanowires ................................................. 66 Figure 3.3. Crystallographic property of bio MO nanowires ..................................................... 67 Figure 3.4. XPS spectra of bio MO nanowires and manganese oxide (MO) nanoparticles...........68 Figure 3.5. Stabilization effect of PAA wrapping on bio MO nanowires................................. 69 Figure 3.6. Pd/Au nanoparticles loading on bio MO nanowires ............................................... 70 Figure 3.7. Elemental mapping of Pd/Au nanoparticles loaded bio MO nanowires...................71 6 Figure 3.8. TGA data and galvanostatic profile of bio MO nanowires and control samples.........72 Figure 3.9. Microstructure analysis of MO nanoparticles and bio MO nanowires .................... 73 Figure 3.10. Li-0 2 battery operation of MO nanoparticles and bio MO nanowires...................74 Figure 3.11. Li-0 2 battery operation of Au, Pd nanoparticle loaded bio MO nanowires electrodes ........................................................................................................................................................ Figure 3.12. The improved specific capacity of Li-0 2 75 battery with the rationally designed catalyst electro des ........................................................................................................................................ Figure 3.13. The improvement of Li-0 2 76 battery cycling performance with the rationally designed catalyst electrodes...........................................................................................................................77 Figure 3.14. The cycling performance of low carbon Li-0 2 batteries with the fixed discharge capacity of 4,000 mAh g-I (400 mAh g-c+catalyst)...........................................................................78 Figure 3.15. The cycling voltage profile of low carbon Li-0 capacity of 2,000 mAh g-Ic (200 mAh g- 2 +catalyst)...........................................................................79 Figure 3.16. The cycling voltage profile of high carbon Li-0 capacity of 727 mAh g-'c (400 mAh batteries with the fixed discharge 2 batteries with the fixed discharge g- c+catalyst)............................................................................81 Figure 4.1. M13 virus mediated synthesis of various Li-0 2 battery materials for Li-0 2 batteries ...................................................................................................................................................... 10 1 Figure 4.2. HRTEM images of biotemplated MCO nanowires.................................................... 102 Figure 4.3. The electrochemical performance of bio MCO nanowires in Li-0 2 batteries ........... 103 Figure 4.4. C haracterizations of N i N Ps....................................................................................... Figure 4.5. The electrochemical performances of bio MnxCo 3 xO4 104 (x=l, 2) nanowires in Li-02 b atteries......................................................................................................................................... 1 05 Figure 5.1 Scheme of Ml 3 virus-based graphene defects detector.............................................. 112 Figure 5.2 Scanning electron microscope (SEM) image of graphene flakes ............................... 113 3 Figure 5.3 p sequences of M 13 virus clones selected from bio-panning against graphene flakes ...................................................................................................................................................... 7 1 14 Chapter 1. Introduction 8 1.1 Background The word to describe the post X-generation has changed from 'Global' generation to 'Tech', 'Digital', even to 'Wii' generation. Thus, storing electrical energy into pockets becomes important and Li ion batteries met the needs to power small portable devices. From 1970s, rechargeable Li ion batteries operating with 'intercalation' mechanism at room temperature' have been studied. Intercalation reactions in Li ion batteries refer to the insertion/de-insertion of Li ion into crystalline phase of active materials. 2 Theoretical capacities of these intercalation materials remain low since the number of electron transferred between the positive electrode and the negative electrode is mostly limited to around one per active 3d metal (e.g. LiCoO 2 , LiFePO 4, LiTiS 2 etc). The amount of energy stored (or dissipated) in batteries is proportional to the number of electron associated in reactions and the cell potential as the following equation shows |AGI= nFE| (n: mol, number of moles of electron passed per mole of reactants, F: 96,485 C/mol, magnitude of electric charge per mole of electron, E: V, potential difference between a cathode and an anode). Thus, finding the battery reaction chemistry accompanying multiple numbers of electrons and a high potential difference is necessary to achieve high energy density batteries. In this thesis, conversion reaction system and Li-oxygen battery system were selected to meet those requirements to develop next generation energy storage devices with high energy densities. 1.1.1 Conversion reaction materials for Li-ion battery cathodes Among the high energy density batteries, the conversion reaction mechanism is promising since each chemical formula of the conversion materials reacts with multiple lithium ions and electrons (MaXb + (b-n)Li -> aM + bLinX). They have great possibilities due to their larger theoretical capacity than the conventional intercalation materials. Compared to other conversion reaction materials including sulfides, oxides, nitrides and phosphides,'0 fluoride materials'' 9 provide high reduction potentials above 2 V vs. Li/Li+, which is applicable to cathodes due to the highly ionic characteristic of metal-fluorine bonds. However, the application of conversion reaction materials has been limited by their small practical capacity because of low conductivity and poor reversibility of electrochemical reactions. Thus, there have been several efforts to increase the conductivity of fluorides. One of the reported methods is to partially replace the fluorine with oxygen' 2 13 to induce covalent characteristic to the compound. For other methods, fluorides have been composited with carbon materials through a high-energy ball milling. With this method, the active materials particle size is reduced to nanoscale thus it has decreased ionic and electronic reactions. diffusion path resulting in the improved reversibility of electrochemical Here, we expect the genetically engineered M 13 virus can improve the power and the reversibility of conversion materials based cathodes. By nanocompositing conducting materials, graphene and bismuth oxyfluorides (BiOO. 5F 2) using M 13 virus, homogeneous graphene based conductive framework that contacts to active materials can result in a significant improvement in the electrochemical performance of a model conversion reaction material, bismuth oxyfluoride. 1.1.2 Li-oxygen batteries for high energy density storage systems Li-oxygen batteries are attractive next generation energy storage systems as they can increase the gravimetric energy density of fully packaged batteries by 2-3 times that of conventional Li-ion cells. 5 In the operation of rechargeable Li-oxygen batteries with nonaqueous electrolytes, Li 2 02is deposited (oxygen reduction reaction; ORR) during discharging and decomposed back into Li+ and 02 (oxygen evolution reaction, OER) during charging at the catalyst electrode (2Li++O 2+2eLi 20 2 ). Although their high theoretical energy density (3,505 Wh kg') is extremely advantageous, current Li-oxygen batteries suffer from a high voltage hysteresis, a low power performance and a short cycle life. These poor performances of Lioxygen batteries were mainly originated from the instability of electrolytes due to the high reactivity of reduced oxygen or discharge products, Li 20 2 .16' 10 17 In addition to electrolytes, the cathodes still remain as a barrier to fully utilize the advantage of Li-oxygen batteries. The catalytic efficiency of Li-oxygen battery cathodes needs to be improved, as they form or decompose highly insulating Li 2 0 2 throughout the electrode during the operation. Furthermore, in recent studies, the cathodes were made with high cost (e.g. Au 18 , Pd, Pt)' 9 or rare (e.g. RuO 20 2 2, Lao. 7 5Sro.2 5 MnO 321) elements, impeding the commercialization of Li-oxygen batteries. Therefore, developing a highly functional and cost efficient catalyst electrode is necessary to achieve high energy density battery systems. In this thesis, the M 13 virus can be applied to form a nanocomposite structure for Li-oxygen battery cathodes with a high surface area and cost efficiency. By developing a synthetic method to homogeneously distribute the functional nanoparticles, the amount of noble metal can be lowered and the surface mediated reaction between lithium ions, oxygen molecules and electrons can be facilitated. 1.1.3 The property of M13 Virus and its electronic device applications This thesis focuses on developing high energy density battery systems with the biomolecule, M 13 virus, to expand the application of batteries into electric vehicles. The M 13 virus is a filamentous bacteriophage, with a length of - 880 nm and a diameter of - 6.5 nm.3 This high aspect ratio virus particle has been used as a nanowire template for the functional inorganic nanomaterials growth such as semiconductors, 4 photovoltaic cells,5 perovskites6 and Li-ion battery electrodes.','8 9 In addition, the single-stranded DNA encapsulated inside the coat proteins can be modified to express specific peptide sequences on the surface of the virus to enhance the interaction with materials of interest. 4' 9 By taking advantages of these two properties of M 13 virus, high energy density battery electrodes can be rationally designed and fabricated with a precise control at the nanoscale. 11 1.2 Scope of work The main focus of this thesis lies on the rational design of nanomaterials for Li-ion battery and Li-oxygen battery electrodes. Nanocomposite structures were suggested with the genetic engineering of M 13 virus by taking into account the reaction mechanism and current limitations of Li-ion battery and Li-oxygen battery. The transportation of two key elements in the operation of battery, lithium ion and electron, was improved by decreasing the active material size into nanoscale and by compositing carbon-based materials or conductive metals. In chapter 2, the M 13 virus was used as a template of BiO0 .5F 2 as well as a stabilizer of graphene. The new M 13 virus clone was selected and further modified, called FC#2, to play the dual functionality in forming Li-ion battery cathodes. The M 13 virus mediated graphene/BiOO. 5F 2 nanocomposites improved the power performance, round trip efficiency of conversion reaction materials, BiOo. F . 5 2 In chapter 3, the synthesis of spherulitic surface morphology of bio-templated manganese oxide (bio MO) nanowires was described. They improved the capacity and cycle life of Li-oxygen batteries at six times higher current density than previously reported manganese oxide based cathodes. Moreover, the surface of bio MO nanowires was further engineered to control the binding affinity with oxygen molecules by decorating a small amount of (3-5 wt % of electrodes) novel metal nanoparticles, Au or Pd. In chapter 4, we were able to easily tune the composition of nanowire with M 13 virus templates, resulting in various transition metal based nanowires, MnCo3 O 4 (x = 0, 1, 2) for the application to Li-oxygen battery cathodes. The different electrochemical behaviors in Li-oxygen batteries were observed with this compositional set of spinel oxides thus enabling us to frame a design principle for future Li-oxygen batteries. In chapter 5, the M 13 virus based graphene defect detector is introduced and its preliminary result is included as ongoing work. 12 1.3 Reference I. Whittingham MS. Electrical Energy-Storage and Intercalation Chemistry. Science 192, 1126-1127 (1976). 2. Whittingham MS. History, Evolution, and Future Status of Energy Storage. P Ieee 100, 1518-1534 (2012). 3. III CFB, Burton DR, Scott JK, Silverman GJ. Phage display: a laboratory manual.Cold Spring Harbor Laboratory Press: Cold Spring Harbor, New York, 2001. 4. Mao CB, Solis DJ, Reiss BD, Kottmann ST, Sweeney RY, Hayhurst A, et al. Virus-based toolkit for the directed synthesis of magnetic and semiconducting nanowires. Science 303, 213-217 (2004). 5. Dang XN, Yi HJ, Ham MH, Qi JF, Yun DS, Ladewski R, et al. Virus-templated selfassembled single-walled carbon nanotubes for highly efficient electron collection in photovoltaic devices. Nat Nanotechnol 6, 377-384 (2011). 6. Nuraje N, Dang XN, Qi JF, Allen MA, Lei Y, Belcher AM. Biotemplated Synthesis of Perovskite Nanomaterials for Solar Energy Conversion. Adv Mater 24, 2885-2889 (2012). 7. Nam KT, Kim DW, Yoo PJ, Chiang CY, Meethong N, Hammond PT. et al. Virusenabled synthesis and assembly of nanowires for lithium ion battery electrodes. Science 312, 885-888 (2006). 8. Lee YJ, Yi H, Kim WJ, Kang K, Yun DS, Strano MS, et al. Fabricating Genetically Engineered High-Power Lithium-Ion Batteries Using Multiple Virus Genes. Science 324, 1051-1055 (2009). 9. Lee YJ, Lee Y, Oh D, Chen T, Ceder G, Belcher AM. Biologically Activated Noble Metal Alloys at the Nanoscale: For Lithium Ion Battery Anodes. Nano Lett 10, 2433- 2440 (2010). 10. Cabana J, Monconduit L, Larcher D, Palacin MR. Beyond Intercalation-Based Li-Ion Batteries: The State of the Art and Challenges of Electrode Materials Reacting Through Conversion Reactions. Adv Mater 22, E170-E192 (2010). 11. Amatucci GG, Pereira N. Fluoride based electrode materials for advanced energy storage devices. J Fluorine Chem 128, 243-262 (2007). 12. Gocheva ID, Tanaka I, Doi T, Okada S, Yamaki J. A new iron oxyfluoride cathode active material for Li-ion battery, Fe2OF4. Electrochem Commun 11, 1583-1585 (2009). 13. Bervas M, Klein LC, Amatucci GG. Reversible conversion reactions with lithium in bismuth oxyfluoride nanocomposites. JElectrochem Soc 153, A 159-A 170 (2006). 13 14. Bervas M, Badway F, Klein LC, Amatucci GG. Bismuth fluoride nanocomposite as a positive electrode material for rechargeable lithium batteries. Electrochem Solid-State Lett 8, A 179-A183 (2005). 15. Bruce PG, Freunberger SA, Hardwick LJ, Tarascon JM. Li-O-2 and Li-S batteries with high energy storage. Nat Mater 11, 19-29 (2012). 16. McCloskey BD, Speidel A, Scheffler R, Miller DC, Viswanathan V, Hummelshoj JS, et al. Twin Problems of Interfacial Carbonate Formation in Nonaqueous Li-O-2 Batteries. J Phys Chem Lett 3, 997-1001 (2012). 17. McCloskey BD, Bethune DS, Shelby RM, Girishkumar G, Luntz AC. Solvents' Critical Role in Nonaqueous Lithium-Oxygen Battery Electrochemistry. JPhys Chem Lett 2, 1161-1166 (201 1). 18. Peng ZQ, Freunberger SA, Chen YH, Bruce PG. A Reversible and Higher-Rate Li-O-2 Battery. Science 337, 563-566 (2012). 19. Lu YC, Gasteiger HA, Shao-Horn Y. Catalytic Activity Trends of Oxygen Reduction Reaction for Nonaqueous Li-Air Batteries. JAm Chem Soc 133, 19048-19051 (2011). 20. Jung HG, Jeong YS, Park JB, Sun YK, Scrosati B, Lee YJ. Ruthenium-Based Electrocatalysts Supported on Reduced Graphene Oxide for Lithium-Air Batteries. Acs Nano 7, 3532-3539 (2013). 21. Xu JJ, Xu D, Wang ZL, Wang HG, Zhang LL, Zhang XB. Synthesis of Perovskite-Based Porous LaO.75Sr0.25MnO3 Nanotubes as a Highly Efficient Electrocatalyst for Rechargeable LithiumOxygen Batteries. Angew Chem Int Edit 52, 3887-3890 (2013). 14 Chapter 2. Integrating graphene based conducting frameworks into Lithium ion battery cathodes (Bismuth oxyfluorides, BiOo. 5F 2) by genetic engineering of M 13 virus* - The content in this chapter was reprinted with permission from Small. Copyright C 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim 15 2.1 Introduction Single-layer graphene sheets have significantly broadened the horizon of nanotechnology with their unique electronic, optical, quantum mechanical and mechanical properties associated with the two-dimensional atomic crystal structure.' To best utilize this material for practical applications, it is crucial to prevent the spontaneous aggregation between individual graphene sheets during composite materials formations. Numerous efforts have been made to stabilize functionalized graphene sheets on molecular2 ' 3 or polymeric species.4' Biomolecules such as DNA 6 and proteins 7 have also been grafted onto graphene planes for applications of biosensors ,8 controlled drug-delivery9 as well as cancer imaging.1' In addition to biomedical applications, graphene sheets can also be hybridized with biomolecules into energy storage devices to increase the conductivity of active materials that are often insulators. In previous work, ultrasonication" or chemical reduction,' 2 followed by heat treatment, 13 ' 14, " have been adopted to achieve composites between graphene and various materials (LiFePO 4 1" and SnO 2 16). However, due to the nonspecific nature of interactions between the graphene templates and active materials, it is expected that only random and inhomogeneous contacts were created, leaving the segregation on nano- or even sub-micron levels. Ideally, the performance of these active materials, such as the accessible capacity and the rate capability, can be maximized only if atomic level contacts can be realized between the conductive phase (graphene) and the active phase. In order to expand the range of graphene based hybrid materials, methods to prevent aggregation around the limit of colloidal stability need to be developed. The stability of aqueous colloidal dispersion of functionalized graphene is usually maintained at high pH and low ionic strength due to the charges on the functionalized surface.' 7 Substrate specificity of ligands in biomolecules can improve the colloidal stability of graphene and strengthen the interaction between graphene and functional materials, thereby providing a genetically tunable hybrid building block and a desired conducting frame. The Ml 3 bacteriophage has been demonstrated as 16 a genetically engineerable biological toolkit to develop nanostructured hybrid materials to enhance the performance of energy storage and conversion devices.'' 19 Here, we show that the non-covalent binding between the engineered M13 virus and graphene increased the dispersion stability of graphene sheets at pH as low as 3 and an increased ionic strength environment. In addition, using this biological approach, we were able to take a DNA sequence that coded for peptides that could bind graphene and modify this sequence to further broaden the stability window of aqueous colloid of graphene sheets. With the improved stability of graphene in aqueous media, inorganic nanoparticles nucleated on the M13 virus were able to intimately interface with graphene sheets and fully utilized the excellent electronic conductivity of graphene, although the incorporation of graphene might lower the packing density. As a result, we achieved an efficient conducting matrix throughout the hybrid material with the genetically engineered M13 virus, which simultaneously stabilizes graphene sheets and mineralizes the active nanoparticles. We also demonstrated that the electrochemical utilization of the originally insulating active materials could be improved in the composite network consisting of active nanoparticles and conductive graphene sheets. 17 2.2 Experimental 2.2.1 Synthesis of bio-templated bismuth oxyfluorides and their composites with graphene FC#2 viruses (2x1013 ) were incubated with the graphene solution (64 ml, 2.3 [tg mlr). Bi(N 3 )3 -5H 2 0 (0.8m1, 200 mM, dissolved in 10% HNO 3) was added to this mixture to make the final solution (400 ml, 0.4 mM). LiF solution (171 ml, 50 mM) was added and stirred at room temperature for 3 hours. Graphene solution (40 ml, 50 [tg mlr) was further added into the nanocomposites. The final products were filtered, washed and dried under vacuum at 50'C overnight. With the same method, wild type viruses were used for bismuth oxyfluoride/graphene nanocomposites and virus addition and incubation steps were excluded for composites without virus. 2.2.2 Synthesis of SWNTs/bio-templated bismuth oxyfluoride nanocomposites The SWNT/FC#2 complexes were constructed by dialyzing the mixture of FC#2 (2x 101) and SWNTs (0.094 mg in 2 wt% sodium cholate aqueous solution) against D.I. water with gradual increase of salt (KCI) concentration of media from 10 mM to 80 mM, while maintaining pH 9 using NaOH for two days. Bismuth oxyfluoride was synthesized by adding Bi(NO ) -5H 0 3 3 2 (0.8 ml, 200 mM, dissolved in 10% HNO 3) to complex solution giving final volume to 400 ml. After an hour, LiF (171 ml, 50 mM) was added and rested for 2 hours. The final products were washed and dried under vacuum at 50*C overnight. 2.2.3 Electrochemical tests with coin cell type batteries Active materials were mixed mechanically with Super P (TIMCAL, SUPER PO Li) for 20 minutes and polytetrafluoroethylene (PTFE) was added. (Active materials : virus : graphene : Super P : PTFE (mass ratio) = 68.8 : 8.8 : 2.4 : 15 : 5). Mixed powders were rolled out and punched in 4.08 mg cm 2 and dried under vacuum at 120'C overnight. Inside the Ar-filled glove 18 box, the electrodes were assembled into coin cells with Li metal foils as the counter electrodes and I M LiPF6 in EMC was used as the electrolyte. Three layers of microporous polymer separator (Celgard 2325) were used. Assembled coin cells were tested with a Solatron Analytical 1 470E potentiostat at room temperature. 2.2.4 FC#2 gene construction The p8cs#3 clone was selected by a bio-panning method with a pVIlI library previously reported by this group.' 9 Using QuikChange Lightning site-directed mutagenesis kit (Stratagene, catalog#210518), K was first mutated to E, and N to D then to E, producing EFE virus. The primer sequences used to change K to E are 5' CAG GGA GTT AAA GGC CGC TTC TGC GGG ATC CGG CAG CGC 3' and 5' GCG CTG CCG GAT CCC GCA GAA GCG GCC TTT AAC TCC CTG 3'; for N to D, primers of 5' GTC GCTGA GGCTT GCAGG GAGTC AAAGG CCGCT TTTGC GGG 3' and 5' CCC GCAAA AGCGG CCTTT GACTC CCTGC AAGCC TCAGC GAC 3' were used; for D to E, primers, 5' GC TGA GGC TTG CAG GGA CTC AAA GGC CGC TTC TGC 3' and 5' GCA GAA GCG GCC TTT GAG TCC CTG CAA GCC TCA GC 3' were used. Previously reported SWNT-binding peptide MC#2,18 was fused to pill of EFE virus. The oligonucleotides used to introduce SWNT-binding functionality to the minor pill coat protein are 5 [Phos]' GTA CCT TTC TAT TCT CAC TCT GAT ATG CCG CGT ACT ACT ATG TCT CCG CCG CCG CGT GGT GGA GGT TC 3' and 5 [Phos]' GGC CGA ACC TCC ACC ACG CGG CGG CGG AGA CAT AGT AGT ACG CGG CAT ATC AGA GTG AGA ATA GAA AG -3' and they were annealed to form a DNA duplex. The cloning vector was extracted from EFE virus using standard miniprep kit (QIAGEN) and was digested with Eag I and Acc65 I enzymes, dephosphorylated, and purified with agarose-gel. Purified vector and DNA duplex were ligated using T4 DNA ligase at 16*C overnight, and electrotransformed to XL-l blue cells. Transformed cells were incubated for one hour, plated, and incubated at 37'C overnight. 19 2.2.5 Synthesis of water-soluble graphene2 Graphene oxide (GO) was synthesized by Hummer's method.2 ' 75 mg of GO was dispersed in 75 ml of water by bath-sonication for one hour followed by centrifugation (3,000 rpm). The pH was adjusted to 10 with 5 wt% sodium carbonate (Sigma-Aldrich) solution. Then 600 mg of sodium borohydride (Sigma-Aldrich) in 15 ml of water was added to partially reduce GO at 80*C for one hour followed by centrifugation (20,000 rpm). After washing with water, the partially reduced GO was dispersed again in 75 ml of water by sonication. The diazonium salt for sulfonation was made with 46 mg of sulfanilic acid (Alfa-Aesar) and 18 mg of sodium nitrite (Alfa-Aesar) in 10 ml of water and 0.5 ml of I M HCl solution in an ice bath. The diazonium salt solution was added to the partially reduced GO in an ice bath for 2 hours. To completely reduce the sulfonated graphene, 2 g of hydrazine (Acros) in 5 ml water was added kept at 100 C for 24 hours. I ml of 5 wt% sodium carbonate solution was then added into the mixture to precipitate the graphene. After centrifuging (25,000 rpm) and washing with water, the lightly sulfonated graphene was dried under vacuum at room temperature for 24 hours. 2.2.6 Characterization of graphene. The water-soluble graphene was characterized by transmission electron microscopy (TEM) in Figure 2.1.a and atomic force microscopy (AFM) in Figure 2.1.b. For TEM, JEOL 2010F TEM was used with accelerating voltage of 200 kV. For AFM, the graphene on mica (Ted Pella) substrate was achieved by drying graphene dispersion and Veeco Nanoscope IV under tapping mode was used. 2.2.7 Zeta potential measurement of virus and graphene. The concentration of virus solution was 1012 m-I in water with 10 mM NaCl. The stock solution of virus (- 1014 m-) was initially dissolved in 10 mM Tris, 15 mM NaCI before diluting in 10 mM NaCl in ddH20. The amount of solution used to generate curve was 30 ml. The ionic 20 concentration of the solution was set to 10 mM NaCl for all samples to minimize the fluctuation of ionic strength during pH adjustment. The pH was then adjusted using NaOH until the pH was around 10. For graphene zeta potential measurement (Figure 2.1.c), the concentration was 40 mg ml' in water and the pH of graphene dispersion was adjusted to around 11 by NaOH. The zeta potential of both virus and graphene were then measured at an accumulation time of 10 with 5 measurements per sample at 20 V using DelsaNano (Beckman Coulter). Electrophoretic mobility was calculated using the Smoluchowski approximation (used for particles larger than 0.2 ptm in I mM or greater salt solution). The pH of solution was then adjusted with HCL. 2.2.8 Characterization of bismuth oxyfluoride and graphene nanocomposites by XRD, XPS and TGA. The crystal structure was confirmed by X-Ray Diffraction (PANanalytical Multipurpose Diffractometer, Cu Ka radiation) and Rietveld refinement was conducted (Table 2.1) by changing the atomic occupancy of oxygen and fluorine. Since oxygen can take a place in fluorine site, we changed the occupancy of fluorine positions (F l: 0.5,0.5,0.5 and F2: 0.25,0.25,0.25) to fit the calculation to the data. For the electrode discharged to 1.5 V, the coin cell was disassembled inside Ar-filled glove box and the discharged electrode was covered by Kapton* tape to prevent any air contamination. X-ray photoelectron spectroscopy (XPS, Kraots AXIS) was conducted with pass energy 20 eV and step size 0.1 eV and the atomic ratio between bismuth and fluorine were measured. To quantify the virus mass of the composites (Figure 2.2.a, b), thermogravimetric analysis (TGA, Q50 TA instrument) was used with increasing temperature with rate of I 0 0 C min" under nitrogen. 21 2.2.9 Characterization of bismuth oxyfluoride and graphene/bismuth oxyfluoride nanocomposite by electron microscope. The bismuth oxyfluoride was characterized by high-resolution TEM (HRTEM) using JEOL 201 OF TEM with an accelerating voltage of 200 kV. The graphene/bismuth oxyfluoride nanocomposites were also characterized by using JEOL 200CX TEM operating at an accelerating voltage of 120 kV. 22 2.3 Results and discussion We utilized an M13 virus to synthesize a graphene/virus complex to function as a building block for a conducting framework. The M 13 virus is a filamentous bacteriophage, with a length of -880 nm and a diameter of -6.5 nm.22 The single-stranded DNA encapsulated inside the coat proteins can be modified to express specific peptide sequences on the surface of the virus to enhance the interaction with materials of interest.23 ' 24 To fabricate a virus-mediated graphene framework with inorganic nanoparticles, two factors must be addressed; the colloidal stability of graphene and the interface between the active materials and the graphene. In designing an M13 virus, the major coat protein (pVIII) was chosen as a major interacting motif to maximize the attraction between the graphene and the virus, so that every particle templated on the virus was forced to contact the graphene (Figure 2.3.a). First, the graphene-binding virus, with an 8-mer peptide insert, DVYESALP, fused to the amino-terminus of the pVIII major coat protein (this virus is called p8cs#3), was identified through a bio-panning method using a pVIlI library previously reported by this group.19 The aromatic residue, tyrosine (Y), of the selected sequence is expected to interact with graphene, as well as single-walled carbon nanotubes (SWNTs), through jT-n interaction. In addition to the aromatic residue, the hydrophobicity plot of the sequence, calculated based on the Hopp-Woods scale with the averaging group size 5, 25 shows a hydrophobic moiety between two hydrophilic regions (Figure 2.3.b inset) suggesting that the virus can bind graphene by hydrophobic-hydrophobic interaction. Second, to facilitate the nucleation of nanoparticles, we introduced two additional carboxyl groups on each pVIII protein of p8cs#3 virus, in which the thirteenth amino acid, lysine (K), and the seventeenth amino acid, asparagine (N), were changed to the glutamic acid (E), using site-directed mutagenesis (Figure 2.3.b top) (this site-mutated virus is called EFE). Since the thirteenth and seventeenth amino acids of an M 13 virus are known to be exposed on the surface and accessible to ligands, 2 6 these carboxyl groups of glutamic acid can chelate metal ions and catalyze the mineralization. The zeta 23 potential of EFE was measured and compared with that of the control virus, p8cs#3, to confirm the effect of the site-directed mutation on the surface charge of the virus (Figure 2.3.b bottom). It was observed that the isoelectric point of the virus shifted to a lower pH by the addition of two glutamic acids on the pVIII protein. The increased negative charges associated with the carboxyl groups have additional advantages in enhancing the colloidal stability of the graphene/virus complex. Finally, the pIII minor coat protein was also engineered to increase the binding affinity between the virus and the graphene (see the experimental section). We designate this virus clone as FC#2 and use it for further research on stabilizing graphene, nucleating functional materials and improving the performance of lithium ion batteries. The stability of the graphene/virus complex was tested by adding bismuth nitrate (the precursor for the material of interest for lithium ion batteries) at pH 3. The stability of graphene dispersion by the virus was maintained after 24 hours of incubation with bismuth nitrate as shown in Figure 2.3.c. This is in contrast to the control sample under the same salt concentration and pH without the virus, where the graphene aggregated. We also observed the graphene/virus (FC#2) complex by atomic force microscopy (AFM) (Figure 2.3.d). The thickness of the graphene in the solution was around 0.8 nm as indicated in Figure 2.3.d inset. An area with the relatively lower coverage of the virus on the graphene compared to the approximate calculation was selected to clearly visualize the interaction of the virus and the graphene. The geometrical quantification of graphene and virus (FC#2) was achieved by following method. The total mass of graphene is 64 ml 147.2 mg. For 2 x 10 1 viruses, there is 147.2 mg / (2 virus, corresponding to 7.36 x 1018 g / 12 g molr x x 6.02 1013) = 7.36 x x x 2.3 mg ml- = 10-" g of graphene per 1021 moP' = 3.7 x 105 carbon atoms. Considering a hexagonal packed structure of carbon atoms, each carbon atom contribute to an area of 0.5 x (0.142 nm) 2 x sin(p/3) x3 = 0.0262 nm 2 . Also considering the graphene thickness is around 0.8 nm, which is close to the thickness of a two-layer graphene, each virus covers an area of graphene of 0.0262 nm 2 x x 3.7 x10 5 =9.7 3 x 10 nm 2 . The cross section area of a virus is 6.5 nm 2 880 nm = 5720 nm 2 . Thus, the coverage of virus on graphene is expected to be 5720 nm / 9700 24 nm 2 = 59%. This calculation is based on the assumption that every single virion of virus binds graphene. Leveraging the enhanced colloidal stability of the genetically programmed graphene/virus complex, we assembled bismuth oxyfluoride on the graphene/virus template. Bismuth oxyfluoride is a conversion reaction cathode material with an open circuit voltage of 2.8 V vs Li/Li+, a high theoretical specific capacity of 210 mAh g- (for BiOo.5F 2, from LiF formation) and an attractive volumetric energy density of 5056 Wh 1-, which can be synthesized in aqueous solution under low pH conditions. Among the candidates of cathode materials for next generation lithium ion batteries, bismuth oxyfluoride was chosen as a model material, since we found the synthetic condition for this material under a weak acidic environment, suitable for demonstrating the improved colloidal stability of the graphene. Since the M13 virus helped maintain the colloidal stability of the graphene during the nucleation of inorganic nanoparticles under low pH, it is now possible to form hybrid nanostructures of the graphene/BiOo. 5 F2 . We first developed an aqueous solution-based approach to synthesize bismuth oxyfluoride nanoparticles on the virus under a weakly acidic condition at room temperature, using LiF as a milder precursor than hydrofluoric acid" and ammonium fluoride. Nanoparticles, thus synthesized along the virus, have a diameter of around 40 nm (Figure 2.4.a,b). The virus-mediated synthesis increased the reaction yield (the mass of final products of each synthesis was measured and the yield of products was calculated as, yield (%) = (mass of BiOO. 5F 2 without virus mass)/(number of moles of bismuth precursor x molecular weight of BiOO. 5F 2 ) x100, thus, the yield was increased from 17% without virus to 63% with virus, both without graphene) and decreased the particle size of the material compared with the previous report.27 To fabricate the nanocomposites of the watersoluble graphene and bismuth oxyfluoride, the graphene was first complexed and stabilized by the FC#2 virus, and then bismuth oxyfluoride was grown on the virus/graphene complex. In order to visualize the hybrid structure with graphene, we used a lowered concentration of the precursor to show that more bismuth oxyfluoride was nucleated along the virus than on the surface of 25 graphene (Figure 2.4.c). The chemically modified graphene possesses functional groups, which also act as nucleation sites, but the graphene-assisted nucleation is not as efficient as the virusassisted nucleation. Two control hybrid materials with the graphene were synthesized without using the virus or with a wild type virus (M13KE, denoted in our work as a wild type virus). In addition to the reduced colloidal stability as shown in Figure 2.3.c, the yield of bismuth oxyfluoride of the reaction without the virus (42%) was much lower than the yield of the reaction aided by the FC#2 (68%). For the nanocomposites made with the wild type virus, the graphene was initially stabilized by non-specific interaction, but the graphene aggregation and the separation between the graphene and active materials eventually prevailed as the nucleation proceeded (Figure 2.4.d). Furthermore, the positive surface charge from the wild type virus under weakly acidic conditions (pH 3) did not facilitate the nucleation of bismuth oxyfluoride and the yield was similar to the reaction without the virus. The crystal structure of the virus-templated bismuth oxyfluoride was confirmed as cubic (Fm3m) by high-resolution transmission electron microscopy (HRTEM) (Figure 2.4.b) and X-ray diffraction (XRD) (Figure 2.5.a), with a lattice parameter of 5.8160(1) photoelectron spectroscopy (XPS) elemental A. From X-ray analysis, the bismuth-to-fluorine ratio was determined to be 1:2.02 (Table 2.2), giving a chemical formula of BiOO.4 9F 2 .02 . The galvanostatic measurement (at a current density of 6 mA g-) further confirmed the composition of fluorine and oxygen based on the fact that oxygen and fluorine in bismuth oxyfluoride react with the lithium at different potentials of 1.8 V and 2.6 V (vs Li/Li+). 2 9 In Figure 2.5.b, the ratio of discharge capacity in the plateau regions around 2.7 V and 1.9 V was calculated to be 1.95:1.05. Because the ethyl methyl carbonate (EMC) electrolyte is believed to be inert to Bi nanocrystals,30 the solid electrolyte interphase formation does not occur under 2 V, therefore does not contribute to the capacity. Since each fluorine atom reacts with one lithium atom and each oxygen atom reacts with two lithium atoms, the galvanostatic measurement gave a chemical formula of BiOo.5 25 F,.95 , in fairly good agreement with the XPS analysis result. Therefore we concluded with a reasonable 26 approximation that the chemical formula of the synthesized bismuth oxyfluoride was close to BiOO.5F2. The advantage of incorporating well-dispersed graphene into lithium ion battery cathodes was demonstrated by making electrodes with the graphene/bismuth oxyfluoride nanocomposites assembled with a biologically engineered virus. With a small amount (2.4 wt%) of graphene incorporated into the bismuth oxyfluoride, the specific capacity of the virus-templated bismuth oxyfluoride was increased from 124 mAh g- to 174 mAh g-1 at a current density of 30 mA g(C/7) (Figure 2.6.a and Figure 2.6.b). The second cycle capacity (206 mAh g-, at C/7) of the virus templated bismuth oxyfluoride with 2.4 wt% of graphene in Figure 2.7.a corresponds to 98% of theoretical capacity, which is the highest reported for this material to date. Moreover, the rate performance has also been improved, showing a specific capacity of 131 mAh g(corresponding to 711 W kg', with an energy density of 316 Wh kg-, 2718 Wh 1-) at a current density of 300 mA g- (1.4 C) compared to 67 mAh g- for the virus-templated bismuth oxyfluoride without graphene. These results represent a significant improvement in the electrochemical performance of bismuth oxyfluoride compared to the previous report, as indicated by the increased active mass loading in the electrode (from 50 wt% to 70 wt%) and the improved specific capacity at a 20 times higher current density. In addition, the presence of the graphene also significantly decreased the voltage hysteresis between the charge and the discharge profiles (Figure 2.7.a), which became more pronounced at a higher discharging rate (Figure 2.6.a). This reduction in the electrode overpotential stems from the excellent electric wiring achieved at the atomic level by graphene sheets, which connects active nanoparticles with the cell current collector through a homogeneously distributed percolating conductive network. To study the beneficial effect of specific interaction between the genetically engineered virus and the graphene in the formation of a conducting framework, control experiments have been done with graphene/bismuth oxyfluoride nanocomposites synthesized in the absence of the virus or in the presence of the wild type virus. The bismuth oxyfluoride synthesized in the control 27 experiments had the same chemical and crystallographic properties as that formed with FC#2 (Figure 2.8 shows XRD patterns for each composite). As shown in Figure 2.6.a, c and d, the specific capacity (174 mAh g-' at C/7) of nanocomposites using FC#2 was higher than that of nanocomposites using the wild type virus (145 mAh g- at C/7) and without the virus (102 mAh gI at C/7). Moreover, the superior electrochemical performance of the nanocomposites using FC#2 was more apparent at a high rate of 600 mA g- (110 mAh g- for FC#2, 67 mAh g- for the wild type virus and 64 mAh g- without the virus), indicating accelerated electrode kinetics for the cell reactions in those nanocomposites benefiting from the specific interaction between the graphene and FC#2 virus. The poor electrochemical activities of the nanocomposites in the presence of the wild type virus and in the absence of the virus are caused by the agglomeration of graphene during the active material synthesis. Based on these observations, we conclude that the increased interaction of graphene with the active material particles aided by the genetically engineered virus results in an efficient conducting network. This was made possible by maintaining the colloidal stability of complex during the synthesis. Moreover, we achieved both higher capacity utilization of the active materials and a much reduced kinetic barrier between charge and discharge reactions, as compared with the previous report.2 9 The superior formation of a percolating network with two-dimensional conducting sheets enabled by the genetically engineered virus has also been demonstrated by comparing the effects of graphene and SWNTs on the electrochemical performance of bismuth oxyfluoride. The SWNTs were evenly dispersed inside the electrode by the virus complexation method' 9 and bismuth oxyfluoride was synthesized on this template. When the same amount of graphene and SWNTs (0.5 wt%) were incorporated into the nanocomposites, the specific capacities of 128 mAh g- for the SWNTs/bismuth oxyfluoride nanocomposites and 181 mAh g- for the graphene/bismuth oxyfluoride nanocomposites were obtained respectively, at a current density of 30 mA g- (Figure 2.6.e). The graphene improves the kinetics of the conversion reaction more effectively than SWNTs. Although the SWNTs are known to be advantageous for constructing a 28 percolating network because of their high aspect ratio,31 when a small quantity of carbon is used, the interconnectivity between the one-dimensional SWNTs could be limited compared to the twodimensional graphene. In the bismuth oxyfluoride system, the graphene appears to be more effective in facilitating the electrochemical reaction. 29 2.4 Conclusion In summary, using a genetically engineered M 13 virus, we broadened the stability window of graphene in aqueous media, which enabled an environment-friendly approach to establish a graphene/virus nanotemplate. By designing the M13 virus for simultaneously stabilizing the graphene and nucleating bismuth oxyfluoride, we fabricated the graphene/bismuth oxyfluoride nanocomposites in which both phases were intimately interwoven. The graphene/virus complex formed a homogeneously distributed conducting framework and the kinetics of electron transfer inside the battery electrodes was improved, demonstrating the increased specific capacity of bismuth oxyfluoride at a high current density (131 mAh g-' at 300 mA g-') and the reduced overpotentials for both charging and discharging cell reactions. This study demonstrates the importance of well-dispersed graphene in aqueous media for synthesizing composite materials and this general method could be extended to other materials for applications including biosensors, supercapacitors, catalysts and energy conversion applications. 30 2.5 Figure and Tables Oil C.1 -25 - >-30* 40. j45- 50. - ---------------- -00. 65. i") 1, pH Figure 2.1. Characterizations of graphene. (a) TEM image of the chemically reduced graphene with a selected area electron diffraction (SAED) pattern. AFM image (b) and Zeta potential data (c) of water-soluble graphene, the size of the AFM image is 2 mm by 2 mm. 31 100 -- 100 - 8W vWpqylIC iss i f loss from vrus a-- 94 -2 - 90 92 Bismuth oxyfluoride 90 Bismuth oxyfluoride-graphene on FC#2, under nitrogen -- on FC#2, under nitrogen 100 200 300 400 500 100 Temperature CC) 200 300 400 500 600 Temperature ( 0 C) Figure 2.2. Determination of virus mass by Thermogravimetric analysis (TGA). (a) TGA data of bismuth oxyfluoride grown on FC#2 showing that virus accounts for 11 wt% of the final hybrid nanowire. (b) TGA data for the analysis of virus mass inside the graphene-bismuth oxyfluoride nanocomposites formed by FC#2 (before adding 2.4 wt% of additional graphene), which shows the similar quantity as bismuth oxyfluoride grown on virus. 32 b pcst3 IAl DI VI YI E S A LPDPA K A A F Site mutation EFE A D YESAL N . 4 C D P A E A A F E . 20 -VVYGSALP 10 02 01 -10 -20 -30 Amino Acl -40 0 N -50 -60 -- EFE p8cs#p -70 3 4 5 6 7 8 9 10 11 10 nm -pH 0 nm 33 Figure 2.3. The graphene/M13 virus complex with the enhanced colloidal stability for the hybrid graphene/nanoparticle nanocomposites. (a) Scheme of the inorganic nanoparticle nucleation on the graphene/virus template. The virus enables a close contact between inorganic nanoparticles and the graphene. (b) Top: The peptide sequence of pVIIl protein of p8cs#3 and EFE. Bottom: The zeta potential of EFE and p8cs#3. Inset: The hydrophobicity plot of the pVIIl major coat proteins of p8cs#3 virus as a function of the amino acid location. (c) The left vial is the graphene solution after 24 hours of incubation with bismuth nitrate, showing the aggregation of graphene on top. The right vial is the graphene/FC#2 complex solution after 24 hours of incubation with bismuth nitrate without any aggregation. (d) AFM image of the graphene/virus (FC#2) complex. Inset: The height profile of line A showing the thickness of graphene as ~0.8 nm. 34 a 500 nm C 300 nm d - -- iif +-Graphone 5 nm Figure 2.4. Characterizations of the bismuth oxyfluoride nucleated on the graphene/virus complex. (a) TEM image of bismuth oxyfluoride nucleated on FC#2. (b) HRTEM image of a bismuth oxyfluoride nanoparticle with the Fm-3m crystal system, (200) d=2.9 (220) d=2.1 A (left arrow) and A (right arrow). (c) TEM image of the bismuth oxyfluoride/graphene nanocomposites with FC#2 virus for battery electrodes. The bismuth oxyfluoride/FC#2 virus complexes are shown exclusively on the graphene. Inset: Catalyzed nucleation of bismuth oxyfluoride on FC#2 virus was observed by lowering the precursor concentration in the composite synthesis with a scale bar of 400 nm. Background is the graphene in the inset. (d) TEM image of the bismuth oxyfluoride/graphene nanocomposites formed with the wild type virus showing the separation of graphene and bismuth oxyfluoride. 35 4.0 a Bismuth oxyftuoride on virus (FC#2) - - Bismuth oxyfluoridelgraphene (2.4 wt%j with FC#2 3.5 03.0 (.4 C LiF formation L~ILf 02.S 00 20 25 30 35 40 45 50 55 0 2.0 Li 2 O formtion C; 60 65 70 2 Theta (degree) 15 0 30 60 90 120 150 180 210 240 270 300 Specific capacity (mAhlg) Figure 2.5. Characterizations of bismuth oxyfluoride with an electrochemical method and XRD. (a) XRD pattern of bismuth oxyfluoride nanoparticles. (b) Galvanostatic data of bismuth oxyfluoride/graphene nanocomposites (2.4 wt% of graphene addition) under constant current density (6 mA g-) until 1.5 V. 36 a3.2 -aj --- 2.8 60 mA/g 150 mA/g' 300 mA/g, --- 00 mAILg --- Bismuth oxyfluoride/graphone b,3. b nooositr 20 40 (2.4 wt%) s with FC#2 60 80 100 120 140160 180 2.8- 2. Bismuth oxyfluorid. on virus (FC#2) 20 40 60 C 3 .2 80 100 120 140 160 180 2.8- 2.4 Bismuth oxyfluoridIelgraphone (2.4 wt%) d 3.2 20 40 60 80 100120140160180 2.8 Bismuth oxyfluoridelgraphone (2.4 wt%) compositswt wild typ virus 3.2 20 40 60 80 100 120 140 160 180 2.0 2.8 Bismuth oxyfluoride/graphenes 0.5 --- 1.6 Bi m t 020 40 xfy 60 80 d*8 t N *05w% 100 120 140 160 180 200 Specific capacity (mAhlg) Figure 2.6. The power performance of the graphene/bismuth oxyfluoride nanocomposites with the genetically engineered M13 virus. (a-d) The first discharge of bismuth oxyfluoride at different current densities; (a) the bismuth oxyfluoride/graphene nanocomposites with FC#2, (b) the bismuth oxyfluoride templated on FC#2 37 without graphene, (c) the bismuth oxyfluoride/graphene composites in the absence of virus, (d) the bismuth oxyfluoride/graphene composites in the presence of the wild type virus. (e) Comparison between the graphene and SWNTs: The first discharge of the bismuth oxyfluoride/graphene composites and the bismuth oxyfluoride/SWNTs composites formed with FC#2 viruses having the same mass percentage of carbon (0.5 wt% of electrodes) at a 30 mA g- of current density. 38 a . . . . oxyfluoridelgraphene 4.8 . . Bismuth 4.4- Bismuth oxyfluoride on FC#2 2.4 b wt% . .,IIeg. * O3.2 t - . . . 50 55 60 Bismuth oxyfluoride-graphene nanocomposite after discharge . Bi metal 2deys nn 2 ycle - 2 cyce 2.48 2.0 0 20 40 60 80 100 120 140 160 180 200 220 2025 Specific capacity (mAh/g) Figure 2.7. Electrochemical 30 35 40 45 65 70 2 Theta (degree) performance and characterization of bismuth oxyfluoride/graphene nanocomposites and bismuth oxyfluoride on FC#2. (a) First (straight) and second (dashed) cycles of bismuth oxyfluoride/graphene (2.4 wt%) nanocomposites with FC# 2 and bismuth oxyfluoride on FC#2 at a current density of 30 mA g-1. (b) XRD patterns of the bismuth oxyfluoride-graphene nanocomposites (2.4 wt% addition) electrode after discharging to 1.5 V with constant current density of 6 mA g-. Bi metal is detected, however, Bi 20 observed and this could be due to small particle sizes.29 39 3 was not a D-- - Bismuth oxyfluoridelgraphone (2.4 wt%) with FC#2 (a=5.8160 A - C Bismuth oxyftuoride/SWNTs (0.5 wt%) with FC#2 (a-5.8213 A) Bismuth oxyftuorld./grsphone (2.4 wt%) without virus (a=5.8158 A -- d - Bismuth oxyfluoridelgraphen. (2.4 wt%) with wild type virus (az5.8145 A) 20 25 30 35 40 45 50 55 60 65 70 2 Theta (degree) Figure 2.8. XRD patterns of control samples. (a) bismuth oxyfluoride/graphene nanocomposites formed with FC#2, (b) bismuth oxyfluoride/SWNTs nanocomposites formed with FC#2, (c) bismuth oxyfluoride-graphene composites formed without the virus, (d) bismuth oxyfluoride-graphene nanocomposites formed with the wild type virus. 40 4 AA4 40 312 25 14 2.0 80 5f Gaphtna w4th F C*2 B 40 2.4 0 0B Graphene w'thOu virus C 4.0 36 28 0 20 40 60890 1001201404601802W0220240 Specific capacity (mAh/g) Figure 2.9. The first five cycles galvanostatic data of bismuth oxyfluoride/graphene (2.4 wt%) nanocomposites oxyfluoride/graphene (BOF-GP) (2.4 wt%) with FC# 2 nanocomposites and control (BOF-GP) samples. with FC#, (A) bismuth (B) bismuth oxyfluoride/graphene (2.4 wt%) nanocomposites without virus, (C) bismuth oxyfluoride/graphene (2.4 wt%) nanocomposites with wild type virus under constant current (30 mA g-). 41 Table 2.1. XRD peaks table for synthesized bismuth oxyfluoride from 20-70 degree (2 Theta) with Rietveld refinement results for compound. (h k I ) 2T(cal) 2T(cor) 2T(obs) Delta d(cal) d(cor) d(obs) Del-d 1% ( 1 1) 26.547 26.547 26.609 0 3.3549 3.355 3.3472 0 100 (2 0 0) 30.748 30.747 30.809 0.001 2.9054 2.9055 2.8998 0.0001 42.2 (220) 44.041 44.042 44.101 -0.001 2.0545 2.0544 2.0518 0 62.7 (3 11) 52.164 52.163 52.221 0 1.7521 1.7521 1.7503 0 55.6 (2 2 2) 54.671 54.669 54.726 0.002 1.6775 1.6775 1.6759 0.0001 16 (400) 64.044 64.038 64.093 0.006 1.4527 1.4528 1.4517 0.0001 7 (cal=Calculated, obs=Observed, cor=Corrected) Table 2.2. Atomic concentration (%) of bismuth oxyfluoride by XPS analysis. Elements Atomic concentration (%) Bi 23.09 F 46.79 42 2.6 References 1. Geim AK, Novoselov KS. The rise of graphene. Nat Mater 6, 183-191 (2007). 2. Bekyarova E, Itkis ME, Ramesh P, Berger C, Sprinkle M, de Heer WA, et al. Chemical modification of epitaxial graphene: spontaneous grafting of aryl groups. JAm Chem Soc 131, 1336-1337 (2009). 3. Petridis D, Bourlinos AB, Goumis D, Szabo T, Szeri A, Dekany I. 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J Electrochem Soc 153, A 159-A 170 (2006). 30. Gmitter AJ, Badway F, Rangan S, Bartynski RA, Halajko A, Pereira N, et al. Formation, dynamics, and implication of solid electrolyte interphase in high voltage reversible conversion fluoride nanocomposites. J Mater Chem 20, 4149-4161 (2010). 31. Landi BJ, Ganter MJ, Cress CD, DiLeo RA, Raffaelle RP. Carbon nanotubes for lithium ion batteries. Energy Environ Sci 2, 638-654 (2009). 45 Chapter 3. Li-0 2 battery hybrid nanocatalyst formed with M 13 virus templated Manganese oxides nanowires and noble metal nanoparticles* - The content in this chapter was reprinted with permission from Nature Communications. Copyright © 2013, Rights Managed by Nature Publishing Group 46 3.1 Introduction Rechargeable Li-0 2 batteries operate through the reversible reaction between lithium ions and oxygen molecules." 2 ' 3 When Li-0 2 batteries discharge in non-aqueous electrolytes, Li' reacts with reduced oxygen (oxygen reduction reaction; ORR) yielding the reaction product, mainly Li 2 0 2 that is deposited on the catalyst electrode at a thermodynamic voltage of 2.96 V (vs. Li/Li+).4'5'6 Upon charging, the electrical energy is stored by decomposing Li 20 2 back into Li+ and 02(oxygen evolution reaction, OER). To promote these ORR and OER for Li-0 2 batteries, several materials have been investigated as catalyst electrodes, such as noble metals (e.g. Au, Pt, Ru, Pd),''' 9 transition metal oxides (e.g. a-MnO 2,'" Co 30 4 ," MnCo 2 0 4 ,'2 RuO 2,") and carbon- based materials (e.g. graphene 415, ' carbon fiber,' 6 doped carbon nanotube' 7 ). However, these materials suffer either from high material costs, low conductivities or side reactions with electrolytes, which limits their use as high performance Li-0 difficulties restrain the capability of Li-0 2 2 battery electrodes. Although these batteries, the rational design and selection of electrode materials may fully deliver these battery chemistries to a high gravimetric energy storage system. To enhance structural functionality, organisms in nature form skeletal tissues from soft to hard through the interactions between nucleating protein matrices and inorganic ions.18, 19,20 Inspired by these biominerals researchers have fabricated novel nanoarchitectures using peptides or proteins as scaffolds applied in various functional structures.2 For example, numerous types of platforms such as synthetic peptide nanofibers,22 nanorings,23 and ferritin cages 24 provide simple and uniform ways to synthesize interesting materials at the nanoscale. In electrochemical energy storage devices, nanostructured materials enhance Li ion batteries by shortening the diffusion length of Li ions, 29 For Li-0 2 25,26,27 and benefit capacitors by providing electrodes with large surface areas. 28, batteries, since the oxygen is in principle derived from the air, both the oxygen flow into electrodes and the high efficiency to promote discharge/charge reactions are additional important factors. 30 Constructing a new catalyst that combines porous nanostructures and high catalytic activity would allow us to harness the advantages of Li-0 47 2 batteries for practical applications, such as long-range electric vehicles. Here, we built the catalyst structures to achieve high Li-0 2 battery performances by forming a nanocomposite of biotemplated manganese oxide nanowires (bio MO nanowires) with incorporation of a small weigh percent (3 wt %) of Pd nanoparticles (Fig. 3.1). We believe this is the first bio-directed synthetic method demonstrated for Li-0 2 battery applications. 48 3.2 Experimental 3.2.1 Synthesis of bio MO nanowires and their nanocomposites with Au or Pd nanoparticles Manganese sulfate monohydrate (MnSO 4-H 20, Sigma-Aldrich, 2.4 ml of 50 mM) and sodium sulfate (Na 2 SO 4 , Sigma-Aldrich, 2.4 ml of 50 mM) solutions were added together into 50 ml of D.I. water. FC#2 viruses (1.63x]01) were added into this solution and stirred at room temperature overnight. After the overnight incubation, potassium permanganate (KMnO 4, Alfa Aesar, 16 ml of 10 mM) solution was added and stirred at room temperature overnight. The final products were washed with D.I. water by centrifugation and collected after lyophilization. To incorporate the noble metal nanoparticles (Pd, Au), the bio MO solution was dialyzed against D.I. water overnight and diluted with D.I. water to make a final solution volume of 200 ml. PAA (MW: 2,000, 0.6g in 20 ml of D.I. water) was added and stirred overnight. The PAA wrapped bio MO nanowires were collected by centrifugation and dispersed again with 240 ml of D.I. water. Au/Pd precursor solutions were added into the previous solution (For Au: gold (III) chloride trihydrate, HAuCl4e3H 20, 2.17 ml of 25.4 mM for 1.7 wt % of Au in the final Au/bio MO nanowire composite electrodes. For Pd: sodium tetrachloropalladate (II), Na 2PdCI 4, SigmaAldrich, 0.612 ml of 34 mM for 3 wt % (0.408 ml of 34 mM for 1.1 wt %) of Pd in the final Pd/bio MO nanowire composite electrodes) and stirred overnight at room temperature. To reduce the metal ion, 3 ml of ethylene glycol was added to each Au/bio MO nanowire and Pd/bio MO nanowire solution for 24 hours inside a brown bottle due to light sensitivity. The solution was washed three times by centrifugation with D.I. water and lyophilized. 3.2.2 Characterizations of MO nanoparticles, bio MO nanowires and Au,Pd/bio MO nanowires For SEM images, FEG-SEM 6700F (JEOL) was used under 5 kV with 5.9 mm of working distance (or 0.5 kV with 3 mm of WD) after coating the sample with Pd. High-resolution transmission electron microscopy (HRTEM) images were obtained with 2100, 2010 FEG (JEOL) 49 under an accelerating voltage of 200 kV. The sample was loaded on formvar/carbon or quantifoil grids (Electron microscopy sciences). X-Ray Diffraction (PANanalytical Multipurpose Diffractometer, Cu Ka radiation) patterns were collected with 0.18 */min scan speeds under the following settings: 20 of anti-scatter slit, 6 mm of irradiated length of automatic mode and 0.04 rad of soller slit. The discharged/charged samples were collected after disassembling batteries inside an Ar-filled glove box and were covered by Kaptono tape to prevent any air contamination. Inductively coupled plasma atomic emission spectroscopy (ICP-AES, Horiba Jobin Yvon ACTIVA-S) was used to determine the atomic ratio between manganese, gold, palladium ions of Au/bio MO nanowires and Pd/bio MO nanowires. The sample was dissolved in nitric acid at 50 C overnight and diluted with D.I. water (ACS reagent grades) to make the final nitric acid concentration of 2 % v/v for ICP measurements. X-ray photoelectron spectroscopy (XPS, Physical Electronics Versaprobe 11) was measured with the monochromatic Al X-ray source (K0, 1486.6 eV) under the pass energy of 23.5 eV and the step size of 0.1 eV. The acquired XPS spectra were calibrated with adventitious carbon peak (Is) as a reference positioned at 284.8 eV. To quantify the PAA mass of the metal nanoparticles/bio MO nanowires composites, thermogravimetric analysis (TGA, Q50 TA instrument) was used with 10 *C min-' increasing temperature rate under nitrogen. 3.2.3 Chemical titration for Mn oxidation state determination The average oxidation state of MO was determined by two separate chemical titration steps reported in the literature. 3 1 The color change indicator (sodium diphenylamine-4-sulfonate, C, 2 HIONNaO 3S, Alfa Aesar) was further added during the titration. MO nanoparticles were used for titrations because the manganese oxidation state is expected to be similar to bio MO nanowires from XPS analysis and the protein residue of bio MO nanowires would cause significant errors during the titration. 50 3.2.4 Electrochemical tests The lyophilized catalyst materials were further dried in a vacuum oven at 90 'C overnight before electrode preparations. The Li-0 2 electrodes were composed of Ketjan black (EC600JD, AkzoNobel), catalyst materials, LiTHionTm dispersion (Ion Power, USA) with the mass ratio of 44 : 36 : 20 for high carbon containing (44 wt %) electrodes and the mass ratio of 8 : 72 : 20 for low carbon containing (8 wt %) electrodes. These three components were dissolved into 2-propanol and mixed overnight using a bath sonicator to get homogeneous slurry mixtures. The mixture was spread on the separator (Celgard, PP2075) by tape casting methods and dried in a vacuum oven at 80'C for 2 hours. The dried slurry was punched into circle electrodes with diameters of 1.27 cm and the mass of each electrode was measured (0.1-0.3 mg/cm 2 ) following further drying in vacuum oven at 80'C overnight before the electrochemical test. Inside the Arfilled glove box, the catalyst electrode was soaked with the electrolyte in advance (0.1 M LiCIO in DME, Novolyte) and assembled into the Li-0 2 4 battery cell designed by our lab. Metallic Li foils (negative electrode), two layers of separators, catalyst electrodes and a positive electrode current collector (316 stainless steel mesh) were sequentially put on the cell and additional electrolyte (100 ptl) was added. After the assembly, the Li-0 2 battery cell was purged with oxygen (Airgas, UPC grade) and closed at 1.08 atm. The purged cell was allowed to rest at least 2 hours before the electrochemical test. The first cycle of Li-0 2 battery cell was tested by galvanostatic measurements at 0.4 A/ge with a Solartron Analytical 1470E potentiostat. For the cycling tests, the pressure of the Li-0 2 battery cell was adjusted again to 1.08 atm after 16 cycles and I A/ge of current was applied. 51 3.2.5 The true electrode surface area specific capacity Here we make estimation in calculating the thickness of the discharge product, Li 20 2. It is assumed that Li 2 0 2 is formed as films throughout the overall catalyst electrode with the volumetric capacity, 2698.6 mAh cm- . First, the true electrode surface area was calculated. The BET surface area of bio MO nanowires is 271.7 m 2 Ketjan black (EC600JD, AkzoNobel) is 1,400 m2 -1 and the reported BET surface area of -1. Thus, the true surface area can be obtained by considering the weight percentage (catalyst: Ketjan black = 36 : 44) of each component in electrode, resulting in 271.7 m 2 g- * 0.36 +1,400 m 2 -' * 0.44 = 713.8 mtrue 2 -c+catalyst. As 13,347 mAh g'c corresponds to 7,340.8 mAh g- c+catalyst, the true electrode surface area specific capacity is -1.0 tAh cm2true. 3.2.6 The Gas Chromatography (GC) of Pd/bio MO nanowires electrodes after the first galvanostatic discharge/charge Two separate Pd (3 wt %)/bio MO nanowires cells were prepared to collect the gaseous sample after discharge/charge steps. After the first galvanostatic discharge under I atm of 02, the cells were disassembled in Ar glove box and purged with Ar to remove 02. These cells were then taken out from the Ar glove box and one of them was charged to 4.15 V at 0.4 A g-1, resulting in the 02 evolution. 250 Rl of gas sample was collected from each discharged and charged cells with an airtight syringe. These samples were analyzed with 7980A GC system (Agilent Technologies) with two sequential separating columns (80/100 Hayesep N (13067-U) and 45/60 molecular sieve (13069-U, Sigma-Aldrich)) and thermal conductivity detector (TCD). The retention time was calibrated with the standard reference sample (H 2 = 0.8 min, 02= 1.1 min, N 2 4.5 min). We observed higher composition of 02 = 1.2 min, CO 2 = in addition to the air introduced during the sample injection in charged cells than discharged cells indicating the 02 evolution reaction from 52 Li 2O 2 oxidations. A small amount of H 2 could be due to the reaction of H 20 with the Li metal anode. 53 3.3 Results and discussion 3.3.1 M13 virus mediated synthesis of manganese oxide nanowires Bio MO nanowires with spherulitic surface morphology were synthesized through a facile and moderate chemical reaction by utilizing the biomolecule, M13 virus. M13 viruses have been used as versatile templates for high aspect ratio nanowire synthesis (e.g. semiconductors,3 2 metal oxides3 3' 34'35) under environmentally benign conditions. Among the five different types of capsid proteins on this virus template, p8 is a major coat protein with 2,700 copies surrounding the single-stranded DNA. In addition to the material specificity of these surface proteins (p8 or five copies of minor proteins existing at one end of virus particle, p3), electrostatic interactions between the opposite charge of precursor ions and functional groups of p8 coat protein broaden the template functionality. In this work, the virus clone with the p8 peptide sequences ADVYESALPDPAEAAFE (named FC#2) 36 was used because it has two additional negative functional groups (-COOH from glutamic acid on 13 th and 1 7 th) than Ml3KE (New England Biolabs) under basic conditions. Using this clone, bio MO nanowires were synthesized by first binding Mn2 ions to the FC#2 virus p8 proteins followed by reacting with KMnO 4 at room temperature. From this simple aqueous biotemplating reaction, homogeneous bio MO nanowires with high aspect ratio (-80 nm in diameter and - [tm in length) were obtained. These nanowires showed spherulitic surface morphology (TEM image in Fig. 3.2.a, b, c). While the flower-like the nucleation of MO along the virus particle spheres of MO were previously synthesized,' developed this spherulitic surface of materials into higher dimensional nanowires. Thus, the hierarchical nanostructures of these bio MO nanowires are expected to be advantageous in Li-0 2 battery performances. While previous studies utilized smooth surface MO nanowires for Li-0 2 battery electrodes 0 ' 39, the rough surface of these bio MO nanowires can provide large catalytic area as well as enough storage space for discharge products. To the best of our knowledge, this is the first report of M13 virus directed synthesis of manganese oxide nanowires, which is 54 applicable not only to Li-0 2 battery electrodes but also to various applications such as electrochemical capacitors 0 , Li ion battery electrodes 4' and water purifications. 4 2 With the aqueous, room temperature approach using M 13 virus templates, multi-valent (Mn 3+/Mn4 ), birnessite-like MO nanowires were developed. The crystallographic and chemical properties of bio MO nanowires were investigated with X-ray diffraction (XRD), high-resolution transmission electron microscopy (HRTEM), the chemical titration method, and X-ray photoelectron spectroscopy (XPS). First, the XRD pattern of virus-templated MO nanowires showed three broad peaks centered at 37', 53.5* and 65.5* (20 of Fig. 3.3.a), partially matching to 8-MnO 2 (birnessite, space group C2/m, powder diffraction file (PDF) no. 01-073-2509) XRD pattern. Also, the surface morphology of bio MO nanowires was similar to that of typical birnessites43, as can be seen in the TEM image (Fig. 3.2.b). The crystal sizes of bio MO nanowires were 2-5 nm and the co-existing amorphous phase was observed by HRTEM (Fig. 3.3.b). The average oxidation state of bio MO nanowires was determined to be 3.51 by chemical titration methods, 3' suggesting a multi-valence state (Mn 3+/Mn4 = 0.96) of nanowires. This mixed-valence property of MO was also observed from XPS spectra in Fig. 3.4. We confirmed that the average oxidation state of bio MO nanowires lies between Mn 3+ and Mn4 by observing two features, the peak position of Mn 2 P3/2 and the multiplet splitting of Mn 3s. First, the Mn 2 P3/2 4± peak position of bio MO nanowires (642.1 eV in Fig. 3.4.a) was close to that of MnO 2 (Mn , 642.2 - 642.4 eV) as reported in the literature.44' 45 However, the width of Mn 3s multiplet splitting (5.2 eV in Fig. 3.4.b) of bio MO nanowires showed a larger separation than that of MnO 2 (4.5 ~ 4.7 eV) but close to that of Mn 2 O 3 (5.2 ~ 5.4 eV)."' 45 Since it is known that the separation between Mn 3s splitting peaks becomes larger as the oxidation state of Mn is lower, 4 6 the larger separations of Mn 3s splitting compared to Mn4 imply the presence of a lower valence state in bio MO nanowires. 55 3.3.2 Nanocompositing ORR catalyst on bio MO nanowires To enhance the ORR activity and surface conductivity of porous bio MO nanowires for Li-0 2 battery electrodes, 1-3 wt % of ORR catalytic metal nanoparticles were homogeneously incorporated onto the surface of the nanowires. It is reported that noble metal nanoparticles (e.g. Pd, Pt, Ru and Au) have higher intrinsic ORR activities compared to carbon with 0.1 M lithium perchlorate (LiCIO 4) in dimethoxyethane (DME) electrolyte.' In particular, very recent studies reported a great stability (Au) 47 and high ORR catalytic activity (Pd)7 in Li-0 2 battery applications. However, those electrodes contained 40-100 wt % of noble metal nanoparticles, making them less practical due to cost efficiency. Thus, we suggest the composite structure of bio MO nanowires combined with noble metal nanoparticles to decrease the loading of precious metals in cathodes while maximizing the catalytic activity of Li-0 2 batteries. The nanocomposites were designed to nucleate the metal nanoparticle on the surface of the bio MO nanowire templates to maximize the contact with discharge products in Fig. 3.1 (red lines, right middle). In addition, the homogeneous distribution of ORR metal catalyst along the bio MO nanowires was important in maintaining the porous structure of the electrodes. To facilitate these requirements, poly acrylic acid (PAA) was incorporated around the bio MO nanowires to nucleate the metal nanoparticle. The PAA wrapped nanowires increased the colloidal stability of the nanowires during the synthesis so that a more homogeneous solution of bio MO nanowires was observed (Fig. 3.5.a). Control experiment without PAA wrapping resulted in free Pd nanoparticle formation and aggregated MO nanowires as shown in the TEM image (Fig. 3.5.b). This composite synthesis method was applied to two different ORR metal catalysts, Au or Pd (Au/Pd). To nucleate these metal nanoparticles on PAA wrapped bio MO nanowires, ethylene glycol was used as a mild and selective reducing agent to react with noble metal precursors (HAuCl 4 , Na 2PdCI 4) slowly, avoiding reactions with virus-templated MO nanowires. 56 This metal nanoparticle incorporation method by stabilizing and trapping precursor ions with PAA is expected to be applicable not only to biotemplated nanowires but also to various nanosized substrates, such as nanofibers. The homogeneous distribution of Au/Pd nanoparticles (with a diameter of 2-6 nm, Fig. 3.6) was observed by elemental mapping using scanning transmission electron microscopy (STEM) in Fig. 3.7. The amount of PAA used to form the composite nanostructure was determined with thermogravimetric analysis (TGA) to be 2.5 wt % of the electrode by comparing the weight loss difference between bio MO nanowires and Pd incorporated bio MO nanowires at 500*C (Fig. 3.8.a). 48 The amount of Au/Pd nanoparticle incorporated in the bio MO nanowires was measured by inductively coupled plasma atomic emission spectroscopy (ICP-AES) analysis and varied approximately 1-3 wt % of the total electrode, depending on the metal precursor amount that was added during synthesis. 3.3.3 Improved Li-0 2 battery performances with bio MO nanowires The virus-templated MO Li-0 mAh/gc+catayst+L202) 2 battery cathodes showed a 9,196 mAh/g, (955 first discharge capacity with 92.6 % of coulombic efficiency at a current density of 0.4 A/ge. The electrochemical tests were performed with 0.1 M LiCIO 4/DME electrolyte under 1 atm of oxygen (further details on measurement methods are described in the experimental section). Although ether-based electrolytes may undergo some side reactions during Li-0 2 battery charging49 , it is one of the most stable electrolytes against the oxygen electrode for Li-0 2 batteries known to date without the reactivity with Li metal like DMSO has.47 Further control experiments with different electrolytes were included in the following section and Fig. 3.8.b. For galvanostatic tests, the operating voltage was limited to 2.2-4.15 V (vs. Li/Li+) to minimize possible parasitic reactions. First, the effect of MO electrode structures on Li-0 2 battery performance was studied by varying the architecture from nanoparticles to bio MO nanowires. Under the same synthetic conditions, MO nanoparticles (60 nm diameter) were synthesized 57 without viruses and their morphology was compared to bio MO nanowires with scanning electron images (Fig. 3.9.a,b). The surface area of both the nanoparticles and microscope (SEM) nanowires was measured by the Brunauer-Emmett-Teller (BET) method giving 98.5 m 2 /g and 271.7 m2/g, respectively. Bio MO nanowires showed a 40 % improvement of Li 2O 2 storage capability (9,196 mAh/gc at 0.4 A/ge) compared to MO nanoparticles (6,545 mAh/ge), as can be seen from the first discharge capacity in Fig. 3.10.a. In addition, the discharge voltage of the bio MO nanowires (-2.68 V) is higher than that of MO nanoparticles (-2.6 V) by -80 mV and the charge voltage of the bio MO nanowires (-3.65 V) is lower than that of MO nanoparticles (-3.77 V) by -120 mV. In other words, the bio MO nanowires showed decreased discharge and charge overpotentials compared to the MO nanoparticles. Since the oxidation state of both the MO nanoparticles and nanowires was determined to be similar by XPS analysis (Fig. 3.4.a, b), the difference in Li-0 2 battery discharge capacity was mainly attributed to their different surface area 5 ' and morphologies. Since MO nanoparticles agglomerate during the synthesis, the low surface area electrode materials (Fig. 3.9.a) limit the surface mediated reaction so that it results in a poor specific discharge capacity. In our biotemplated system, the high aspect ratio as well as larger surface areas (Fig. 3.9.b) formed highly active Li-0 2 battery cathodes where the electrode spaces can be fully utilized while minimizing the effect of discharge product blocking the electrodes. The major discharge product obtained after the first discharge of bio MO nanowire electrodes was Li 2 0 2 confirmed by XRD in Fig. 3.10b. The XRD data of discharged electrode matches well to the reference XRD pattern of Li 2 0 2 (orange star, PDF no. 01-074-0115). There was no other major discharge product observed except Li 2 O 2. In previous reports, MOs have been shown to produce a LiOH phase during the discharge 39' residues. The Li-0 2 49 due to the structural water or hydroxyl battery performance of dehydrated (thermal treatments at 160*C for 2 hours) bio MO nanowires was investigated to eliminate any possible contributions from water molecules or hydroxyl groups. The thermally treated nanowires showed a similar discharge capacity of 58 9,206 mAh/ge at 0.4 A/g, (Fig. 3.8.b) compared to nanowires without heat treatments, although structural water decreased from 3.2 to 1.5 wt % of electrodes after heat treatment (Fig. 3.8.a). Thus, the LiOH formation is negligible in this Li-0 3.3.4 The Li-0 2 2 battery performance. battery performance with different electrolytes The first galvanostatic cycle of bio MO nanowires was tested with I M lithium triflate (LiCF 3SO 3) in tetra (ethylene) glycol dimethyl ether (TEGDME) to investigate the Li-0 performance dependence on electrolytes. Here, bio MO nanowire electrodes 2 with LiCF 3SO 3/TEGDME exhibited higher charging overpotential as well as lower first discharge capacity (Fig. 3.8.b, red, 8,766 mAh g'e at 0.4 A g-e). Furthermore, the coulombic efficiency of nanowire electrodes dropped from 92.6 % (0.1 M LiC1O 4 in DME) to 61.5 % (1 M LiCF 3SO 3 in TEGDME) at the same charging cut off voltage, 4.15 V. Since increasing the test voltage windows higher than 4.15 V may evoke the electrolyte decomposition, DME was suitable for bio MO nanowire electrodes in order to achieve higher coulombic efficiency in lower voltage ranges. 3.3.5 Li-0 2 battery performances of Pd/Au loaded bio MO nanowires The hybrid nanostructure composed of Pd nanoparticles and bio MO nanowires exhibited improvement of the Li-0 2 battery discharge capacity. The first galvanostatic cycle of Au/bio MO nanowires and Pd/bio MO nanowires were compared with approximately the same amount of Au (1.7 wt %) and Pd (1.1 wt %) incorporations in each nanocomposited electrode. After the addition of only -I wt % of Au or Pd nanoparticles, the first discharge capacities of Au/bio MO nanowires and Pd/bio MO nanowires (cut off voltage at 2.2 V vs Li/Lie) were improved by 8 % (Au) and 15.5 % (Pd) respectively, compared to bare bio MO nanowires (at 0.4 A/gc, Fig. 3.11.a, Au: green straight, Pd: red dashed). In addition, the Pd/bio MO nanowire electrodes showed approximately 60 mV higher discharge voltage than Au/bio MO nanowires as can be observed from Fig. 3.11.a. The higher discharge voltage of Pd/bio MO nanowires than Au/bio MO nanowires in the first 59 galvanostatic cycle of Li-0 and oxygen. 2 electrodes may be attributed to the attractive interaction between Pd Also, although Pd has a lower contrast than Au on TEM observation due to its lower atomic mass, large (higher than 5 nm) Pd nanoparticles on bio MO nanowires were rarely observed than Au nanoparticles decorated bio MO nanowires (Fig. 3.6) so the smaller nanoparticle distribution of Pd/bio MO nanowires in a given synthetic conditions may improve the battery performances. Thus, the surface electrocatalytic functionality of bio MO nanowires was easily tuned by the composite synthesis method developed here. The capability to store lithium peroxide was further improved by increasing Pd incorporations from 1.1 wt % to 3 wt % of Pd/bio MO nanowires electrodes. With 3 wt % of Pd incorporation, a 45 % improvement of the first specific capacity of bio MO nanowires (Fig. 3.11.a, red straight, 13,347 mAh/gc, 1,008 mAh/gc+cataystLi20 2 ) was achieved at 0.4 A/gc. This is the highest specific capacity under a high applied current (0.4 A/ge, 0.22 A/gc+catalyst) among all manganese-oxide-based electrodes reported to date.39' 3 The formation of Li 2 0 2 after discharge and its decomposition during the charge (oxygen evolution was detected by Gas Chromatography (GC) in Table. 3.1) with Pd/bio MO nanowire electrodes was also confirmed by Raman spectroscopy in Fig. 3.11.b. No significant amount of side product (e.g., Li 2 CO 3) can be detected by XRD and Raman, suggesting that DME is largely stable during discharge. We believe the enhanced discharge capacity of Pd/bio MO nanowires originated from the better utilization of porous electrode spaces by improving the electrochemical active surface area and surface conductivity of bio MO nanowires. The Pd nanoparticles located on the surface of the bio MO nanowires facilitated ORR reactions and improved the electronic conduction throughout the Li 2 02 layers allowing further discharge product depositions. The differences in the charging profile of the 3 wt % Pd-MO nanowires compared to the bare bio MO nanowires (Fig. 3.10.a) can be associated with the increased thickness of Li 2 02 layer of the 3 wt % Pd/bio MO nanowires (-4 nm Li 2 0 2 ) compared to the bare bio MO nanowires (-2.6 nm Li 2 O 2 ) due to larger discharge capacity. However, the charging potential of Pd/bio MO 60 nanowires after decomposing 0.05 IAAh/cmtre 2 of Li 2O 2 was still -100 mV lower than that of catalyst-free, carbon electrode55 at the same depth of discharge, -1 rtAh/cmtme2 (the true electrode surface area specific capacities, the detail calculation method is the experimental section). This lower OER potential compared with catalyst-free electrodes suggests the catalytic function of Pd/bio MO nanowires even after the deep discharge in Li-0 2 battery. The following (second) cycle profile of MO nanoparticles, bio MO nanowires, Au (1.7 wt%)/bio MO nanowires, Pd (1.1 wt%)/bio MO nanowires and Pd (3 wt%)/bio MO nanowires are included in Fig. 3.12.a. The second charging profile showed enhanced performance than previous manganese-oxides-based electrode at -6 times higher current density3 9 and further studies to improve the cycle life after the deep discharge remains as future work. To clarify the role of Pd nanoparticles, PAA wrapped bio MO nanowires were tested as Li-0 2 battery electrodes under the same test conditions. The first discharge capacity of the PAA/bio MO nanowires electrode was 9,722 mAh/gc (Fig. 3.8.b, green), which is only a 5 % improvement of bio MO nanowire electrodes. This small improvement of discharge capacity could be attributed to the decreased aggregation between bio MO nanowires after PAA stabilization. The average value of discharge specific capacity can be seen in Fig. 3.12.b. Therefore, the increased first discharge capacity of Pd/bio MO nanowire composites originated from Pd nanoparticle incorporation onto bio MO nanowires, not from the PAA wrapping. We expect that this methodology can be easily applied to other promising ORR or OER candidate materials to develop efficient catalyst structures for Li-0 2 batteries in future studies. 3.3.6 Lowered carbon amounts in Pd/bio MO nanowires cycling electrodes The catalyst nanostructure designed in this work, Pd/bio MO nanowires, allowed for an electrode with greatly decreased carbon content (8 wt %) in the catalyst electrodes to improve cycle life of Li-0 2 battery. Although carbon functions as a conducting agent and an ORR catalyst, it can also reduce the cycle life because side products can be formed at the carbon/electrolyte or 61 carbon/Li 2O 2 interface in electrodes with ether-based electrolytes.56 By reducing carbon amounts in our electrodes, cycle life of Li-0 A/gc+catalyst) 2 batteries was enhanced at high current densities (1 A/ge; 0.1 even under relatively low charging cutoff voltage (4.15 V). The cycling tests were conducted by limiting the discharge capacity to 2,000 mAh/g, (200 mAh/gcatalyst) or 4,000 mAh/gc (400 mAh/gc+cataIyst, 299 mAh/gc+catayst+Li202) following the previous works method.:' ' Similar to the first discharge specific capacity, the cycle life of Li-0 2 batteries was gradually enhanced as the catalyst structure changed from MO nanoparticles to nanowires and finally to composite Pd/bio MO nanowires (Fig. 3.13-16). In particular, the OER overpotential was lowered as the surface area of catalyst electrodes as well as the surface conductivity of bio MO nanowires was improved by Pd incorporation (Fig. 3.13.a) after being discharge to 4,000 mAh g-'c (400 mAh g-'c+catalyst). Thus, the first cycling round trip efficiencies of MO nanoparticles, bio MO nanowires and Pd/bio MO nanowires was improved from 75 %, 77.7 % to 79.2 %, respectively, as the cathode was rationally designed. The aggregated microstructure of MO nanoparticles limited the Li-0 A/ge (0.1 2 battery cycle life to 10 cycles with 4,000 mAh/gc (299 A/gc+cataiyst). mAh/gc+catayst-Li2o2) at I The bio MO nanowires showed cycle retentions up to 20 cycles due to the high porosity and large catalytic surface area of the electrodes. After incorporating Pd nanoparticles, the discharge capacity of Li-0 2 batteries maintained stable to 50 cycles (Fig. 3.13.b, 3.14). When the depth of discharge capacity was fixed to 2,000 mAh/ge (at I A/ge; 0.1 A/gc+catalyst), these electrodes showed a longer cycle life with a similar cycling performance trend. Each MO nanoparticles, bio MO nanowires and Pd/bio MO maintained the 2,000 mAh/gc of discharge capacity up to 22, 47 and 58 cycles, respectively (Fig. 3.13.b, 3.15). The reversible cycling number with Pd/bio MO nanowires electrodes is smaller than the recently reported,3' 47 which might be due to the instability of electrolyte or the reactivity between carbon and oxygen reduction intermediates.56 , 7 This can be observed from the charging cycle profile in Fig. 3.14-16, resulting in the faster decay of charging capacity than the discharging capacity as cycling number 62 increased in Fig. 3.13.b. Ongoing and future work will be focused on developing stable electrolytes and carbon-free electrodes to enhance the cycling of these electrodes. To investigate the effect of carbon in cycle of Li-0 2 batteries, electrodes were prepared with 44 wt % of carbon. The cycle life of bio MO nanowires and Pd/bio MO nanowires decayed faster than those of low carbon electrodes under the same fixed discharge capacity (728 mAh/ge, 400 mAh/g+catajys,) and current density (I A/ge, 0.1 A/gc+catayst). The observed cycle number was 17, 21 and 30 for MO nanoparticles, bio MO nanowires and Pd/bio MO nanowires, respectively (Fig. 3.16). Although the electronic conductivity was higher for 44 wt % versus 8 wt % carbon electrodes, side products can be formed in the high carbon electrodes during cycling 56 counteracting the kinetic advantage in Li-0 2 battery performance. Therefore, we believe Pd/bio MO nanowires are a viable materials platform for long lasting, high capacity Li-0 63 2 batteries. 3.4 Conclusion Using a biological template, a new catalyst electrode was developed that increased capacity (13,350 mAh/gc; 7,340 mAh/gc; 400 mAh/gc+catajyst at 0.4 A/gc) and cycle life (50 cycles with 4,000 mAh/gc+catalyst at I A/gc; 0.4 A/g+catayst) of Li-0 2 batteries at the highest gravimetric current density compared with other transition metal oxide based electrodes in literatures.' 39 These templates allowed the formation of high aspect ratio manganese oxide nanowires wrapped in 3 wt % Pd nanoparticles facilitating compositional control that outperforms materials made by mechanical mixing.; These high aspect ratio bio MO nanowires formed a porous network that maximizes the interaction between the catalyst and the discharge products (Li 2 0 2 ) thus increasing reversibility and generating a stable cycle life of Li-0 2 batteries. Furthermore, the Pd/bio MO nanowires allowed the implementation of low carbon electrodes batteries improving battery cycle life. We believe this template is readily adaptable for other materials combinations and can provide a platform to test various catalytic electrode materials for Li-0 2 batteries and other catalytic systems. In addition, the application of biomaterials will broaden the material selection scope into diverse areas in designing Li-0 2 development of energy storage systems. 64 battery electrodes thus contributing to the 3.5 Figure and Tables Biotemplating MnOx Carbon particle M13 virus 8I ADVYESALPDPAEAAFESL.. Porous biotemplated NWs electrodes 02 PAA wrapping on biotemplated MnO NWs Incorporation of metal NPs Figure 3.1. Schematic of a nanocomposite Porous and catalytic biotemplated NWs electrodes structure. Synthesis step of the metal nanoparticle/M 13 virus-templated manganese oxide nanowires (bio MO nanowires) and the operational reaction inside Li-0 2 battery cells. 65 a C - 200 nm Figure 3.2. Electron microscope images of bio MO nanowires. TEM images of Ml 3 virustemplated MO nanowires (a) at low magnitude (b) zoomed at the end of an individual bio MO nanowire or the whole nanowire (c). 66 Biotemplated MnO NWs AL 10 20 30 40 50 2 Theta (degree) 60 70 Figure 3.3. Crystallographic property of bio MO nanowires. (a) XRD pattern of bio MO nanowires with the weak and broad peak signals due to the poorly crystalline phase of nanowires. The orange triangle marks (2 theta at 37*, 53.50, 65.50) correspond to the lattice spacing observed from HRTEM FFT image (Fig. 3.2b). (b) HRTEM image showing amorphous phase mixed with crystalline phase (Inset: the Fast Fourier Transform (FFT) pattern with two major rings matching to the atomic planes with the spacing of 2.4 A and 1.6 A, respectively). 67 a - 2 b Mn 2p, Mn 2p Mn 3s 5.2 eV Mn 2p. C C b~ W~ C MnOx NPs Bkoemplated MnOx NWs 660 656 652 PnOx NPs Boemplaed MnOx NWs 648 644 640 636 96 94 92 Binding energy (eV) 90 88 86 84 82 80 78 76 Binding energy (eV) C Bomplated MnO. NWs C 040 C 1,100 1.00 900 800 700 600 500 400 Binding energy (eV) 300 200 100 0 Figure 3.4. XPS spectra of bio MO nanowires and manganese oxide (MO) nanoparticles. The high resolution spectra of bio MO nanowires at (a) Mn 2p region, (b) Mn 3s regions showing 5.2 eV of 3s multiplet splitting. (c) The full range survey XPS spectrum of bio MO nanowires indicates that no element other than Mn is present within the bio MO nanowires. 68 Figure 3.5. Stabilization effect of PAA wrapping on bio MO nanowires. (a) The digital camera image of PAA wrapped bio MO nanowire solutions (left) and bare bio MO nanowire solutions (right). (b) TEM image of bio MO nanowires composited with Pd nanoparticles without PAA wrappings. 69 Figure 3.6. Pd/Au nanoparticles loading on bio MO nanowires. HRTEM image of (a) Pd/bio MO nanowires showing high contrast round shape particles (Pd) with 2.2 A of lattice spacing that corresponds to (111) planes of Pd. (b) Au/bio MO nanowires with dark round particles (Au) having 2.3 A of lattice spacing that corresponds to (I 11) planes of Au. 70 I Figure 3.7. Elemental mapping of Pd/Au nanoparticles loaded bio MO nanowires. (a) STEM image of Pd/bio MO nanowires (left) and the corresponding Energy Dispersive X-ray (EDX) elemental mapping images of manganese (Mn, red, upper right) and palladium (Pd, green, lower right, scale bars, 100 nm). (b) STEM image of Au/bio MO nanowires (left) and the corresponding EDX elemental mapping images of manganese (Mn, red, upper right) and gold (Au, yellow, lower right, scale bars, 50 nm). 71 0 Specific capacity (mAh/gc) 2,000 4,000 6,000 8.000 10.000 a100 4.0903.5 800 3.0 2 70- only Biotemplated MnOx NWs Mno - 2.0 Biotemplated MnOx NWs with heat treatments Biotemplated MnO NWs with PAA/Pd NPs 50 100 0 500 300 400 200 Temperature (celsius) 1.5 Biotemplated MnOx NWs 1M LiCF3SO 3 TEGDME Heat treatment PAA wrapping 0 600 Ketjan black 1,000 2.000 3000 4.000 5,000 6,000 Specific capacity (mAhlgc+catalyst) Figure 3.8. TGA data and galvanostatic profile of bio MO nanowires and control samples. (a) TGA curve of MO nanoparticles, bio MO nanowires, bio MO nanowires after heat treatment at 160*C for 2 h and Pd/bio MO nanowires. Since there is almost no difference in weight percent 59 between MO nanoparticles and bio MO nanoparticles at 500 C , the mass percentage of virus in bio MO nanoparticles is negligible. In addition, 1.623x 10"3 of virus particles (one virus particle weighs ~17 MD) were consumed to produce 24 mg of bio MO nanoparticles. Therefore, (2.82x10- 4 x 1.623x10" )/24 x100%=1.9 %, which is 0.68 % of mass in electrode thus we can almost neglect the mass percentage of virus in Li-0 cycle (at 0.4 A g-', under I atm of 02) 2 battery electrodes. (b) The first galvanostatic of bio MO nanowires tested with the electrolyte composed of I M LiCF 3SO 3/TEGDME (red), bio MO nanowires after heat treatment at 160*C for 2 h (blue, with 0.1 M LiCIO 4 /DME) and the PAA wrapped bio MO nanowires (green, with 0.1 M LiClO 4/DME). 72 a BET surface area: 271.7 m2 /g BET surface area: 98.5 m2/g Figure 3.9. Microstructure analysis of MO nanoparticles and bio MO nanowires. The SEM image and BET surface area of (a) MO nanoparticle showing bulky phase of aggregated particles (Scale bar corresponds to 1 pm) and (b) homogeneously dispersed bio MO nanowires with larger pores (scale bars, I pm). The two BET surface areas are 98.5 and 271.7 m 29-, respectively. 73 a 0 Specific capacity (rnAhlgc) 2.000 4,000 6.000 8,000 10.000 - U.I. 4.5 4.0Charged 3.5 S3.0 25 **D ed Biotemplated MnO NWs MnO NPs 2 0 - ~ Biotemplated MnO NWs electrodes 1.5 . 0 1,000 2000 3000 4,000 5.000 Specific capacity (mAh/gc+catalyst) Figure 3.10. Li-0 2 2 Theta (degree) battery operation of MO nanoparticles and bio MO nanowires. (a) The first galvanostatic Li-0 2 battery cycle of MO nanoparticles (green) and bio MO nanowires (red) at a current density of 0.4 A/ge (-0.044 mA/cm 2 ) with 0.1 M LiClO 4/DME electrolytes under I atm of oxygen. (b) XRD patterns of bio MO nanowire electrodes at the pristine state (blue), the end of the first full discharge until 2.2 V (green) and the end of the first full charge until 4.15 V (red) tested under 0.4 A/ge of current density. The XRD data of discharged electrode matches well to the reference XRD pattern of Li2 0 2 (orange star, PDF no. 01-074-0115). There was no other major discharge product observed except Li 2 O 2 . 74 a Specific 0 3.000 4'5 capacity (mAh/gc) 9,000 12.000 6,000 15.000 - I - Biotemplated MnO. NWs nanocomposites I 40 3.5- Dischargled Charged Charged D pak G peak 2.52. 0-0 20 --PAA/Au NPs (1.7 wt%) PAA'Pd NPs (1 1 wt/a) PAA/Pd NPs (3 wt%) o 2,000 4,000 6,000 Specific capacity (mAhlgc+catalyst) Figure 3.11. Li-0 2 8000 800 1.000 1,200 1,400 1.600 Raman Shift cm 1 . 1800 battery operation of Au, Pd nanoparticle loaded bio MO nanowires electrodes. (a) The first galvanostatic cycle of bio MO nanowires with the similar weight percent of the different type of metal nanoparticle composites (Pd nanoparticles: red dash, Au nanoparticles: green straight) tested under the current density of 0.4 A/g. Further improvement was observed with 3 wt % addition of Pd NPs (red straight) onto bio MO nanowires. (b) Pd (3 wt %)/bio MO nanowires electrodes after the discharge (red) and charge (blue) tested under the current density of 0.4 mA g-1, with 0.1M LiCIO 4/DME electrolyte. 75 a Specific capacity (mAhigc) 4 5 -. 4 0 0 3,000 6:000 9000 12.000 15.000 b 000 1000 12-000 1.0001 3 > -8.000 S4 000 =2.5 ,0 C O NPS B", * i Botempiated MiO NWs 2.0 2.000 PAAAw NP, 11.7 wtyBrto MO NWs NPs 1 1wtV iOMO NWs NP- 13 wto MO NWs U -- -PAAPd -PAAPd 0 2,000 4.000 6,000 Specific capacity (mAh/9c+catatyst) 4.000 2,000- 0 0 Biotempt ted Btemp[ 8,000 Biotemplated d Mn0 NWs MnO, NWs/PAA MnO NWs with Figure 3.12. The improved specific capacity of Li-0 2 PAAPd NP, battery with the rationally designed catalyst electrodes. (a) The second galvanostatic cycle (at 0.4 A g-' under I atm of 02) of various electrodes MO nanoparticles (green), bio MO nanowires (blue), PAA/Au NPs (1.7 wt%)/Bio MO nanowires (yellow), PAA/Pd NPs (1.1 wt%)/Bio MO nanowires (red dash) and PAA/Pd NPs (3 wt%)/Bio MO nanowires (red straight) were tested with the electrolyte, 0.1 M LiC1O 4 /DME. These are the following second cycles of the first cycle of Fig. 3.10a and Fig. 3.11a. (b) The specific capacities of electrodes with MO nanoparticles (orange), bio MO nanowires (blue), bio MO nanowires with PAA wrapping (green), Pd/bio MO nanowires (red) including standard deviations error bars (154-854 mAh/ge). Each sample type was made and tested at least triplicate. 76 a Specific capacity (mAh/g.) 1.000 0 2.000 3,000 b 4.000 45 500~-- 2. . Eectrodes with 8 wt % of carbon Electrodes with 8 wt % of Carbon )uI oi 2 4004 4.0- 4.000 300 3 > 3,0- -2,000 MnOX NPs 25- MnOX NWs Bonemplated MnOX NWs with PAA'Pd NPs 0 S 0 -~-Betemplated -- 0 200 300 400 Specific capacity (mAh/gc+catalyst) 100 Boterp afed MnO 10 1.000 rrr MnO NWs -~-MOps. Bolwmplated 20 NWs w, PAA!Pd NPs 30 40 Cycle number 50 60 0 70 Figure 3.13. The improvement of Li-0 2 battery cycling performance with the rationally designed catalyst electrodes. (a) The first cycling voltage profile of Li-0 2 batteries with the fixed discharge capacity of 4,000 mAh/g, (400 mAh/gccatalys,) for MO nanoparticles (blue), bio MO nanowires (green) and Pd/bio MO nanowires (red) at the current density of I A/ge. Each electrode was composed of 8 wt % of carbon and was cycled in 2-4.15 V voltage windows. (b) The cycling performance of Li-0 2 battery electrodes composed of MO nanoparticles (blue), bio MO nanowires (green) and Pd/bio MO nanowires (red) at the current density of I A/ge (0.1 A/gc+catalyst) with the two different fixed discharge capacities of 2,000 mAh/g, and 4,000 mAh/g, (200, 400 mAh/g+catalyst). The circular shape indicates discharge capacities (for both, 2,000 and 4,000 mAh/ge), the triangle indicates charge capacities (until 4.15 V) for 2,000 mAh/ge and the inverted triangle indicates charge capacities for 4,000 mAh/gc. Each electrode was composed of 8 wt % carbon. All of electrochemical tests reported in this work were repeated for at least six different devices. 77 4.5- 4 Specific capacity (mAhjg) 1.000 2.000 3000 4.000 0 Electrodes with 8 wt of carbor 5,000 - -J' cycae 0 35- '03,0 > 0 25 2.0 MnOx NPs 0 b 45. 0 100 200 300 400 Specific capacity (mAhigc+caataysI) 500 Specific capacity (mAh/gc) 1000 2,000 3,000 4,000 Electrodes with 8 *t of carbon 5,000 c -- 10 cyde '41 Cycle 335 -6 3,0 5 30 20 0tm ated Mo0 NWs 0 C 0 45 100 200 300 400 Specific capacity (mAhgqc+catatys I 500 Specific capacity imAhigci 1,000 2.000 3,000 iectrodes witn 8 W" of carbon 4.000 5,000 (;yde - 1() 2r 40 . '13' y cycle 50' cycle o3 20 .BoKempIeodMno NWs with PAA]PO NPs 0 100 200 300 400 Specific capacity (mAh/gccatayt) 500 Figure 3.14. The cycling performance of low carbon Li-0 2 batteries with the fixed discharge capacity of 4,000 mAh g~'c (400 mAh g'c+catayst). (a) MO nanoparticles, (b) bio MO nanowires and (c) Pd/bio MO nanowires at the current density of I A g-'c. Each electrode was composed of 8 wt % of carbon and was cycled in 2-4.15 V voltage windows. 78 a Spebftc cpartly (mArlgc ) 500 1.000 1,500 2.000 0 4.5 of caboi Electodes mdh 8 1M - 4,0 > 2 500 1 - i c0ycle -- 35 S3.01 20- Mno NP, 50 0 100 Specific capaCdty 150 200 Specic capac y (mAqC) 1500 1000 500 0 wctodes wh S wt I^f 250 (mmh,'C,4rays arbxor 2.500 2.000 1 -- eyde 36 5 2 WCn a SpoI 0 C ),Mo 50 100 1 0 Specfir capacity (mAhIy,+, 0 CaOacity 500 1000 NWb, 200 250 , yst) 4 OTI(mAh 0 P 1,500 2.000 2.50,) 4.5 ctrodes E wi 8 w-Ii o cadx C 20 225 2V0 Bvte 0 + iiated MnO. 51 250 NNs wth PAA PO 100 150 NPs 200 250 2,500 E 5. ectrcdt,,,, *d 8 wt %of rcarbor 200 150 2.000 L0 B0 tempatea MnirO, NWs -- jotemptateduno, NWs wePAAPd NPs 0 to 2' 300 15 50 60 Cyrte numftr Figure 3.15. The cycling voltage profile of low carbon Li-02 batteries with the fixed discharge capacity of 2,000 mAh gc (200 mAh g~' +,tyt).(a) MO 79 nanoparticles, (b) bio MO nanowires, (c) Pd/bio MO nanowires at the current density of 1 A g~',. Each electrode was composed of 8 wt % of carbon and was cycled in 2-4.15 V voltage windows. 80 ImAh/gc) 500 600 700 8W0 Specific capacity a4 .5 0 100 200 300 400 with 44 WF% of carbon ElecIrodoa 900 -cycl 4,0 0 3.014 2.5 2.0-' MnO NPs 0 500 100 200 300 400 Specific capac4y (mAh!gc+calatlyst Specific capacity (mAh/g.) b 100 200 300 400 500 600 700 800 900 0 Electrodes wfth 44 W % of carbon 1 cycle 10 Cycle 20 cyde 403+5- 3,0- -6 2 0- B.empadf 0 C WnOW NW% 200 300 400 Specific capac;ty (mAh!gc+catalys) 500 100 Specific capacity (mAh/c) 100 200 300 400 500 600 700 800 900 0 4.5 Electrodes with 44 W of carbon iy'.yde -- 20 cyce cycle ___30 cxde 10 40 4 0 ~3Q. 20 1 oafnplated MnOx NW wi th PAA(Pd NPs 0 100 d 200 300 400 500 Specific capablty mAhgC ctalyst) 500 Electrories with 44 Wi - %1 800 of Co 400. 6(00 ,00 400 200 U10 NW'. Brk+iplaled MnO, NW' wilh PAA;Pd NPs - 0 200 e nO AP Biolerrplated MnO, - , 0 - - - - - 10 20 Cycle number - - - - 0 30 81 Figure 3.16. The cycling voltage profile of high carbon Li-0 2 batteries with the fixed discharge capacity of 727 mAh g-' (400 mAh g~'c+catalyst). (a) MO nanoparticles, (b) bio MO nanowires, (c) Pd/bio MO nanowires and (d) cycle life of MO nanoparticles, bio MO nanowires and Pd/bio MO nanowires at the current density of I A g-c. Each electrode was composed of 44 wt % of carbon and was cycled in 2-4.15 V voltage windows. 82 Table 3.1. The Gas Chromatography (GC) data of Pd (3 wt %)/bio MO nanowires electrodes after the first galvanostatic discharge/charge at 0.4 A g~', with 0.1 M LiClO in 4 DME. Sample name Retention Time (min) Gas Type Bio MO NWs with PAA/Pd NPs (3 wt%) discharged Bio MO NWs with PAA/Pd NPs (3 wt%) charged 0.801 1.127 1.274 H2 02 ~4.5 CO 2 0.794 1.119 1.270 H2 02 N2 -4.5 CO 2 Area % 1.594 28.682 69.722 Not detected 3.716 36.251 60.031 Not detected N2 Area ratio normalized with N2 0.022 0.411 1 0.061 0.603 1 - Table 3.2. The discharge/charge capacity (mAh g~',) for the first and second cycle of different electrodes galvanostatically tested at 0.4 A g-' with 0.1 M LiClO 4 in DME under 1 atm of 02. Sample name MnOx NPs Biotemplated MnOx NWs Bio MO NWs with PAA/Au NPs (1.7 wt%) Bio MO NWs with I" Discharge Is Charge capacity capacity capacity capacity 6695.1 9196.5 9935.5 4524.8 8516.5 7196.5 5781.6 10528.8 11264.1 1853.3 4367.4 3268.9 10620.1 8549.9 13378.2 3328.4 12267.6 _ 3276.7 2 nd Discharge 2 "dCharge PAA/Pd NPs (1.1 wt%) Bio MO NWs with PAA/Pd NPs (3 wt%) 13347.2 9839.1 _ _ 83 _ 3.6 References 1. Abraham KM, Jiang Z. A polymer electrolyte-based rechargeable lithium/oxygen battery. JElectrochem Soc 143, 1-5 (1996). 2. Christensen J, Albertus P, Sanchez-Carrera RS, Lohmann T, Kozinsky B, Liedtke R, et al. A Critical Review of Li/Air Batteries. J Electrochem Soc 159, R I-R30 (2012). 3. Jung HG, Hassoun J, Park JB, Sun YK, Scrosati B. An improved high-performance lithium-air battery. 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Pd/MnO2 Air Electrode Catalyst for Rechargeable Lithium/Air Battery. Electrochem Solid St 13, A165-A 167 (2010). 59. Lee YJ, Yi H, Kim WJ, Kang K, Yun DS, Strano MS, et al. Fabricating Genetically Engineered High-Power Lithium-Ion Batteries Using Multiple Virus Genes. Science 324, 105 1-1055 (2009). 88 Chapter 4. Investigation of catalytic behavior of M 13 virus templated MnxCo.O 4 02 battery cathodes 89 (x = 0, 1, 2) nanowires for Li- 4.1 Introduction The practical energy density of future battery pack can be improved to ~1,000 Wh kgwith the non-aqueous Li-0 2 battery system.' This potential improvement is promising, since the current Li-ion battery can only deliver 100~140 Wh kg-' (-100 miles of driving range with a single charge), impeding the replacement of gasoline powered vehicles (-300 miles of driving range with a refueling). 2 Despite of this attractive energy density, the current state of non-aqueous Li-0 2 batteries is preliminary. They suffer from the large voltage hysteresis, low power, poor cycle life and the absence of efficient filter that selectively passes oxygen molecules. Many recent studies achieved significant improvements with a novel approach to solve those limitations of Li02 battery systems. Some researchers applied a redox mediator to achieve 100 cycles at the high current density3 (at I mA cm 2 ) and the other utilized the gel cathode/solid catholyte structure to cycle the Li-0 2 battery (at 0.16 mA cm 2 ) for 100 times in ambient air 4 . In addition to employing these advances in Li-0 2 battery systems, a systematic study to select good cathode materials among various compounds is also important to develop a better performing Li-0 Although there are many reports focusing on a single material as a Li-0 Nao. 4 4 MnO 2 , 5 Lao.75Sro.2 5MnO 3, 6 TiC, 7 Ru0 8 2 2 2 battery. battery cathode (e.g. ), direct comparisons between different materials were hardly made. Therefore, a versatile tool to screen the possible candidate for an efficient Li02 battery cathode is required to produce a high energy density storage system. Spinel oxides have shown many interesting physicochemical properties with a diverse combination of elements, AB 2 0 4 (A: metal cation occupying tetrahedral sites, B: metal cation occupying octahedral sites) applicable to magnetism,9 electrocatalyst' 1112 system. 1 and energy storage In particular, cobalt manganese spinel oxides (MnCo 3.,0 4, x = 0-3, here we designate them as MCO) have presented considerable catalytic activities for oxygen reduction reaction (ORR) and oxygen evolution reactions (OER) in alkaline solutions. 3 ' 14 These spinel oxides were also studied in previous reports as a cost effective electrocatalyst for Zn-airl and Liair batteries but with carbonate based electrolytes 90 5 which decompose during Li-air battery operations.1 6 Here we first report the biomolecule directed synthesis of MCO to investigate their functionality as Li-0 2 battery cathodes with ether based electrolytes. While the conventional synthetic methods generated random sizes of particle'', plate' and spheres' 9 of MCO, the Ml3 virus enabled us to form homogeneously distributed high-aspect ratio MCO nanowires (25-50 nm of diameter and ~1 tm of length). Thus, we could observe the correlation between the Li-0 2 battery performance and the chemical composition of bio MCO nanowires while confining the geometry of cathode materials in a similar scale. M13 bacteriophage is a filamentous virus with a single-stranded DNA encapsulated by various major and minor coat proteins. It is approximately 880 nm long and 6 nm in diameter (Fig. 4.1). The viral capsid is composed of 2,700 copies of helically arranged major coat proteins, p8, and 5-7 copies of p3, p6, p9 and p7, located at either ends of the M 13 virus. The genetic sequence of these proteins can be modified to display short (<8-12 amino acid) peptide sequences at their own locations on the M 13 virus. In particular, the E3/E4 virus clone (having a sequence of AEEEE (E4) or AEEE (E3) at the N-terminus of each p8) was used as an effective 2 versatile template for the nucleation of a wide variety of metal oxides such as Co 30 4 ,' , 20 perovskite oxides (SrTiO 3, BiFeO 3).21 Since negatively charged virus surfaces make them easily interact with cationic metal precursors, metal oxide nanomaterials were formed with a high production yield. We recently synthesized MnOx nanowires for Li-0 2 battery cathodes using a M 13 virus clone ADVYESALPDPAEAAFE at p8, presenting negatively charged virus surfaces originated from Aspartate (D) and Glutamate (E). 22 In this study, we extended the bio-directed synthesis of Li-0 2 battery cathodes to different metal cations (Co, Mn) with their diverse stoichiometries (Fig. 4.1) using a versatile clone, E3/E4. 91 4.2 Experimental 4.2.1 Biotemplated synthesis of CoO, nanowires The Co precursor solution was prepared by sequentially adding D.l. water (14 mL), NH3-H 20 solution (4 mL, 28 %) and hydrogen peroxide (2 mL) solution into a glass flask with 10 mmol of cobalt (II) acetate tetrahydrate ((CH 3COO) 2 Co-4H 2 0, 98 %, 500 mM) solution with a vigorous stirring. This Co precursor solution (0.5 mL, 500 mM) was added slowly into the E3/E4 virus solution (150 mL, l.Ox101 pfu mUl, pH 9). After stirring this solution at room temperature more than 6 hours, NH 3 -H20 solution (1 mL, 10%) was added and stirred for one hour. Hydrogen peroxide solution (0.1 mL, I M) was added and reacted for one hour at room temperature. The temperature of this solution was increased to 60'C and was stirred for 12 hours. The final product, M 13 virus-templated cobalt oxide nanowires with the smooth surface morphology, was obtained. The as-synthesized biotemplated cobalt oxide nanowire solution was washed with D.I. water by centrifugation, at least repeating twice. 4.2.2 Biotemplated synthesis of Mn,Co 3 ,0 4 (x=1, 2) nanowires Aqueous Co, Mn precursor solutions were prepared by mixing the cobalt (II) sulfate heptahydrate aqueous solution (CoSO 4 -7H 2 0, >99%, 500 mM), manganese (II) sulfate monohydrate aqueous solution (MnSO 4 -H 20, >99%, 500 mM) in Mn:Co molar ratio of 2:1 and 1:2 for CoMn 2O 4, MnCo 20 4, respectively. For bio-MC20 NWs: 0.32 mmol of Co and 0.16 mmol of Mn, for bio-CM20 NWs: 0.16 mmol of Co and 0.32 mmol of Mn was added slowly to E3 virus solutions (100 mL, 5.Ox1012 /mL) with vigorous stirring. The solution was kept stirring at room temperature more than 2 hours, and then NH 3*H 20 solution (5%) was added to adjust the pH of solution between 9 and 10. The color of solution quickly changed from light red to light green and gradually evolved to dark brown with the time for bio-CO NWs. For bio-MC20 and CM20 NWs, the color of solution changed from light red to yellow brown instantly. After one hour, hydrogen peroxide solution (H 2 0 2 , I M, 0.48 mL for bio-CO NWs, 0.32mL for bio-MC20 92 and CM20 NWs) was added while stirring at room temperature for 2 hours. To increase the yield and rate of bio-CO NWs formation, the solution can be further heated up to 80 0C in oil bath with stirring. The final product can be collected by centrifugation at 3,300 rpm. The product was washed at least twice, with 70 v/v % ethanol and span at 3,300 rpm for 20 min. 4.2.3 Synthesis of Ni nanoparticles The synthesis was carried out under oxygen free environment. In a typical synthesis of Ni NPs, 0.1 mmol of nickel (II) acetylacetonate (Ni(CH 3COCHCOCH 3) 2(H20) 2, 95%, Ni(acac) 2 ) and 0.3 mmol of Tert-butylamineborane (TBAB, dissolved in 2 mL of oleylamine (OAm, >70%)) were dissolved in a mixture of oleic acid (OA, 90%, 0.2 mL) and OAm (10 mL) at room temperature with stirring. The temperature of this mixture was maintained to 160 'C for 30 min under vacuum to dehydrate the precursor and remove trace water in solvents. Under argon gas flowing over the solution in flask, the temperature of solution was further increased to 270 'C with a constant heating rate of 15 'C min-, and kept for 20 min. Then, the solution was cooled down. A mixture of 3-mercaptopropionic acid (6 [l) and hexane (0.5 mL) was injected to the solution under stirring when temperature went below 90 'C. The final product was washed with ethanol and separated by centrifugation (10,000 rpm for 15 min), and then dispersed in ethanol. 4.2.4 Bio CO/Ni nanoparticles nanocomposite formation Bio CO/Ni nanoparticles nanocomposites were obtained by binding the preformed metal nanoparticles on the surface of cobalt oxide nanowires. The bio CO nanowire solution (100 mL) and Ni nanoparticles dispersion (in ethanol, or ethanol/water, 1.8 mmol) was mixed overnight. 93 The weight percent of Ni in nanocomposites was calculated as 1.76 wt %. The final product was washed with water and collected with centrifugation. 4.2.5 Characterizations of biotemplated MnCo3 ..,0 Biotemplated MnCo 3.,0 4 (x=O, 1, 2) nanowires 4 (x=0, 1, 2) nanowires were imaged with high-resolution transmission electron microscopy (HRTEM, 2010 FEG, JEOL) under an accelerating voltage of 200 kV. X-Ray Diffraction (PANanalytical Multipurpose Diffractometer, Cu Kca radiation) patterns of bio MCO nanowires were collected with 0.41 '/min scan speeds under the following settings: 2' of anti-scatter slit, 6 mm of irradiated length of automatic mode and 0.04 rad of soller slit. The surface areas of bio-MCO nanowires were measured with Brunauer-Emmett-Teller (BET) method (Micromeritics, ASAP 2020) after one hour of degassing at 50 'C. 4.2.6 Electrochemical tests The final products of biotemplated MnCo 3 .,0 4 (x=0, 1, 2) and bio CO/Ni nanoparticles nanocomposites were lyophilized and further dried in a vacuum oven at 90 C overnight. The Li02electrodes were prepared with the same method described in chapter 3. The spinel oxide based electrodes were made with Ketjan black (EC600JD, AkzoNobel), catalyst materials, LiTHionTM dispersion (Ion Power, USA) with the mass ratio of 44 : 36 : 20 or 8 : 72 : 20. These three components were mixed with 2-propanol using a bath sonicator overnight. This mixture was tape casted on the separator (Celgard, PP2075) and dried in a vacuum oven at 80*C for 2 hours. These electrodes were punched into circle with diameters of 1.27 cm and the mass of each electrode was about 0.1-0.3 mg/cm 2 . Before the electrochemical test, these electrodes were further dried in vacuum oven at 80'C overnight and were soaked with the electrolyte in advance (0.1 M LiC1O 4 in DME, Novolyte) inside the Ar-filled glove box. Li-0 2 batteries were assembled with metallic Li foils (negative electrode), two layers of separators, additional electrolyte (100 [d), catalyst 94 electrodes and a current collector (316 stainless steel mesh) sequentially. After taking the cell from Ar-filled glove box, the Li-0 2 battery cell was purged with oxygen (Airgas, UPC grade) and closed at 1.08 atm. The cell was rested at least 2 hours before the electrochemical test. The galvanostatic test was conducted at 0.4 A/ge with a Solartron Analytical 1470E potentiostat. In cycling tests at 1 A/ge (100 mA/g+catalyst) of current, the pressure of the Li-0 adjusted again to 1.08 atm after 16 cycles. 95 2 battery cell was 4.3 Results and discussion Biotemplated MCO nanowires were synthesized by two-step reactions, decoration of precursor metal ions along the M 13 virus and oxidation of these ions afterwards. First, the E3/E4 virus solution was incubated with a stoichiometric mixture of metal cation precursors, Co2+ and Mn2+. The biotemplated MCO nanowires were formed along the M 13 virus after the hydrolysis and condensation with the addition of ammonia solution or H20 2 while adjusting pH from 8 to 10. The bio MCO nanowires were characterized by High-resolution transmission electron microscopy (HRTEM) (Fig. 4.2.A-D). We observed the particle shape growth of Co 3 O 4 /Co(OH) 2 (designated as CO) after the oxidation of Co 2 + stained virus (Fig. 4.2.A). While there was no significant crystalline peak observed for bio CO nanowires synthesized at room temperature, they changed to crystalline phase of bio Co 30 4 (Co 30 4 PDF no. 00-043-1003, cubic) nanowires after annealing at 400 *C for 10 minutes (Fig. 4.2.E). The annealing temperature was adjusted to be long enough for further crystallization but not too long to destroy nanowires morphology. Furthermore, the BET surface area of biotemplated C030 4 nanowires increased from 59 to 90.2 m2 /g after annealing. This increase of surface area might be originated from the roughened surface of bio Co 30 4 nanowires after heat treatment as can be observed by TEM images (Fig. 4.2.A,B). The surface roughness of bio MCO increased as Mn ion involved during the nanowire nucleation. Incorporation of Mn during the synthesis developed nanosheet structures along the biotemplate (Fig. 4.2.C, D), resulting in the additional surface area of nanowires. While as synthesized bio MnCo 3.,0 4 , (x = 0, 1) nanowires were amorphous or poorly crystalline, CoMn 20 4 showed perfect crystalline phase matching to the reference card (CoMn 2O 4 PDF no. 00-055-0685, tetragonal) (Fig. 4.2E). The surface area of bio MnCo 204and bio CoMn2 O 4 was measured with BET method, showing 104 and 95.6 m 2 g', respectively. The Li-0 2 battery performance of bio CO and heat-treated bio C030 4 nanowires was investigated with ether based electrolyte, 0.1 M LiClO 4 in DME. The specific capacity of bio CO nanowires was improved by 30 % after thermal treatments of bio Co3O 4 nanowires, from 11,000 96 mAh/gc (6,050 mAh/gc+catalyst) to 14,340 mAh/gc (7,890 mAh/gc+cata 1 yst) (Fig. 4.3.A). These Li-0 batteries were tested at the current density of 400 mA/ge (220 mA/gc+catalyst) 2 with the custom designed cell having closed pure oxygen (Airgas, UPC grade) environment at I atm. To the best of our knowledge, this is the highest specific capacity achieved under highest gravimetric current density with Co 3 0 4 based Li-0 2 battery electrodes up to date. In addition to the improvement of specific capacity, heat-treated bio C0 30 4 nanowires showed higher coulombic efficiency (91.5 %) than as synthesized bio CO nanowires (83 %) within the relatively lower cut-off voltage windows (2.2-4.15 V, vs Li/Li+) than other works. These improvements in electrochemical performance of Li-0 2 battery after thermal treatment of bio Co 30 4 nanowires could be originated from the increased surface area (from 59.9 to 90.2 m2 /g) of catalyst materials. High surface area electrodes provide enough electrocatalytic reaction sites thus resulting in a higher capacity as well as a better coulombic efficiency. Here we further functionalized bio CO nanowires by hybridizing preformed Ni nanoparticles on the surface of nanowires. The successful synthesis of Ni nanoparticles was confirmed with HRTEM showing around 5 nm of their diameter and XRD, as can be seen from Fig. 4.4. After the incorporation of Ni nanoparticles on bio CO nanowires, the specific capacity of Li-0 2 battery was slightly improved to 11,830 mAh/gc (6,500 mAh/gc+catajyst for bio Ni/CO nanowires) and 14,830 mAh/gc (6,500 mAh/g.catayst for bio Ni/CO nanowires after heat treatment). More interestingly, the charging profile of Ni composited electrode was different from that of bio CO nanowires based electrodes. While a single step charging profile was observed for bio CO nanowires electrodes, two flat voltage regions around 3.3 V and 3.9 V were observed for bio Ni/CO nanowires electrodes during charging (Fig. 4.3.B). Since the amount of discharge product for bio CO nanowires and bio Ni/CO nanowires was approximately similar, the different charging profile might be resulted from the surface mediated electrochemical reaction not from the different thickness of discharge products. In Fig. 4.3.A, the charging voltages after decomposing 500 mAh/gc of discharge products were 3.34 V (for bio CO nanowires) and 3.31 V 97 (for heat treated bio Co 30 4 nanowires). We observed the decrease of charging voltages after 500 mAh/gc of Li 2O 2 decomposition for Ni composited bio CO nanowires electrodes, 3.18 V and for heat treated bio Ni/CO nanowires, 3.24 V in Fig. 4.3.B. Therefore, these decreases of charging potential after the incorporation of Ni nanoparticles in electrodes can suggest a better electrocatalyst design for Li-0 2 battery electrodes. The improved OER activity after incorporating Ni nanoparticles on bio CO nanowires was more clearly observed when the same amount of discharge product was oxidized (Fig. 4.3.C). At the given thickness of Li 20 2 (8,000 mAh/gc corresponding to -2.5 nm of Li 2O 2 if it formed homogeneous layer on catalyst electrode), heat-treated bio Ni/CO nanowires showed a lower charging potential than bio CO nanowires, heat-treated bio Co 30 Ni/Co 30 4 4 nanowires, and bio nanowires. Since the overall surface area of catalyst electrodes would be similar after heat treatment or Ni nanoparticle decoration, the lowered charging profile of heat-treated bio Ni/CO nanowires might be due to the improved surface conductivity with Ni nanoparticles incorporation. The better OER activity after heat-treatments than as synthesized Ni/bio CO nanocomposites could be originated from the improved contact between Ni nanoparticles and bio CO nanowires with annealing. In addition, heat-treated Ni/bio CO nanowires showed a longer cycle life maintained to 32 cycles than that of bare bio C0 30 The cycling performance of Li-0 mAh/gc (200 The mAh/gc+catayst) 4 nanowires, 17 cycles (Fig. 4.3.D). battery electrodes was tested with a fixed capacity of 2,000 at I A/ge (100 mA/ge). electrochemical composition (MnCo 3.0 2 4 performance of biotemplated spinel , x=0, 1, 2) was investigated as Li-0 2 oxides with different battery electrodes. As the morphology changes to urchin-like shape with the addition of Mn to bio CO nanowires, the specific capacity of bio MnCo 2 0 4 and CoMn 20 4 nanowires was larger than that of bio CO nanowires by 14-17 % (Fig. 4.5.A), which might be originated from the improved surface area of bio MCO nanowires. Moreover, the charging profile of bio MCO nanowires evolved in two steps while bio CO nanowires showed only a single step during charging. While bio CoMn 204 98 nanowires showed a larger sloppy region than bio MnCo 2O 4 nanowires during charging in the first full cycle, both bio MCO nanowires showed similar charging behaviors when they were charged after cutting the discharge capacity to 8,000 mAh/ge (Fig. 4.5.B). Although the charging behavior of bio MnCo 2 0 4 and CoMn 20 4 nanowires was similar, bio MnCo 20 presented a longer cycle life to 16 cycles than bio CoMn 20 4 4 nanowires nanowires which could be cycled only 10 times with a fixed capacity of 2,000 mAh/g, (200 mAh/g+catalyst) at I A/ge (100 mA/ge) (Fig. 4.5.C). 99 4.4 Conclusion We formed a various compositional set of spinel oxides, MnxCo 3.xO 4 (x=0, 1, 2) nanowires by using biomolecule, the M 13 virus. All of these three different cobalt manganese oxides can be simply synthesized by two-step reaction and they shared the similar geometrical scale around -50 nm of diameter, -1 [tm of length. We observed two different nanowire morphology evolutions with each chemical component, the particle shape growth with cobalt and the planar shape growth with manganese along the M 13 virus. These cobalt manganese spinel oxides were applied to Li-0 2 battery cathodes to investigate their electrocatalytic behavior. The modification of these nanowires with thermal treatments, Ni nanoparticles incorporations showed improvements of Li-0 2 battery performances. This biologically formed nanowires suggest an easy route to frame the selection criterion for high performance Li-0 100 2 battery cathode materials. 4.5 Figures and tables 0 0 0 0 0 C0304 000* M13 virus 0 x= 2,3 0 MnCo 20 4 0 Oxygen * Manganese * Cobalt CoMn 2 O4 Figure 4.1. M13 virus mediated synthesis of various Li-0 2 battery materials for Li-0 2 batteries. Through the interaction between transition metal ions (Co+, Mn+) and the major surface protein of M13 virus (left side), here we synthesized three different compositions of MnCo 3.,0 4 (x = 0, 1, 2) nanowires to utilize them as Li-0 101 2 battery electrodes. E Bio Co 0 NWs Bio Co.O4 NWs after heat treatment o CO 3 .(00-043-1003) -Bio MnvCo, O, NWs S,000231237) Bio CoMn2qO NWs ACOWn,0 (0"-55-M65) 20 30 40 50 60 70 2 Theta (degree) Figure 4.2. HRTEM images of biotemplated MCO nanowires. The scale bars in A-D correspond to 50 nm. (A) bio CO nanowires, (B) heat-treated (400 *C for 10 minutes) bio C0 30 4 nanowires, (C) bio MnCo 204nanowires and (D) bio CoMn 2O4 nanowires. (E) XRD patterns of bio CO nanowires (green), heat-treated (400 *C for 10 minutes) bio C0 30 4 nanowires (violet), bio MnCo 2O 4 nanowires (blue) and bio CoMn2 O4 nanowires (orange). 102 Specific capacity (mAh/gc) A45 0 6,000 3,000 9,000 Specific capacity (mAh/gc) 12,000 15000 B 0 8,000 9000 12M0 15,000 45 40- 40 35- 35 3.31V 3.34 V$ P3 3000 3.24 V M 3.0- 3.18 V 3.02: 5 2.5Bi C030 4 NWs - BK C030 4 NWs after heat treatment 20- 0 2,0 -- 2,000 4.000 6,000 8,00 Specific capacity (mAh/gc+catalyst) - Bo Ni.Co 3 0 4 Nws Ba NiCo3 0 4 NWs after heal treatment 0 4.000 2000 6000 8.000 Specific capacity (mAh/gc+catalyst) Specific capacty (mAh/gcI C4 5 0 2.000 4000 6.000 8,000 I D 2500 250 Electrodes with Ketjan 8 wt % 4,01 (/, 2000 -200 E 1500 150 0h 3.5 8 3 3.0 4 1000- 100 > 1o Cc 3 0 4 NWs 2.51 500.U s -- L ,o0 0 -- iA O m 3 4 Nwls aftr h"I irii"ine Ni co30 4 NWW after heat ieatmni 1.000 2.000 3.000 4.000 ) BiO Co 3O4 NWs after heat treatment Ba Ni/Co 30 4 NWs after heat treatment 5 10 15 20 Cycle number Specific capacity (mAhIgc+catalyst) 25 30 Figure 4.3. The electrochemical performance of bio MCO nanowires in Li-0 C) The first galvanostatic cycle of Li-0 2 2 -50 0 35 batteries. (A- batteries tested with 0.1 M LiC104 in DME electrolytes within a cut-off voltage 2.2-4.15 V at the current density of 400 mA/ge under I atm of 02. The discharge/charge profile of (A) bio CO nanowires (dashed) and heat-treated (400 *C for 10 minutes) bio C0 30 Ni/Co 30 4 4 nanowires (straight), (B) bio Ni/CO nanowires (dashed) and heat-treated bio nanowires (straight). (C) Voltage profiles of Li-0 2 batteries tested with the fixed discharge capacity (8,000 mAh/ge). (D) The discharge capacity (fixed with 2,000 mAh/ge;200 mAh/g+catalyst) against cycle number for bio Ni/CO nanowires (orange circle) and heat-treated bio Ni/Co 30 4 nanowires (blue triangle) tested at 1 A/ge (100 mA/g+atalyst) under I atm of 02. The electrodes were formed with 8 wt % of carbon and 72 wt % of catalyst materials. 103 B -- Ni (00-004-0850) ~M5 - Ni NPs ' 20 30 40 50 2 Theta 60 70 Figure 4.4. Characterizations of Ni NPs. (A) the HRTEM image and (B) the XRD pattern of Ni NPs. 104 A 0 45 Specific capacity (mAhtg) 3,000 6.000 9,000 12,000 15,000 B 45= 0 Specific capacity (mAh/g.) 2000 4,000 6,000 8.000 4.0 3.5 3.5 3.0 3.0 2.5 Bio0Co U NWs 2.5- Rio MnCo 0 NWs - Bi CoMn 0, NWs 2.0 0 2,000 4, )00 6,000 Specific capacit) (mAhIg, ) C 2. 500 20 8,000 4--.... .....- 2200 NWs Bio MnCo Bia CoMn O,NWs O,NWs 1,000 2,0 DO 3,000 4,00 Specific capac , I Electrodes with Ketjan 8 wt % 2, 000 - - Bio CoO I 250 -200 .. 0 E 500- 150 U:) 2. 000 100 "C tF 1. 5O0 -- 0 50 Rio MnCo,O nanowires u nZ Bio CoMnO, nanowires 3 12 I0 15 18 Cycle number Figure 4.5. The electrochemical performances of bio MnCo3 .0 4 (x=1, 2) nanowires in Li- 02 batteries. (A) The first galvanostatic cycle of bio CO (yellow), MnCo 2O 4 (straight pink) and bio CoMn 20 4 (straight green) nanowires at 400 mA/&. (B) The Li-0 2 batteries voltage profiles of bio MnCo 2O 4 nanowires (dashed pink) and bio CoMn 20 4 nanowires (dashed green) tested with the fixed discharge capacity (8,000 mAh/ge). (C) The cycle life (fixed with 2,000 mAh/g; 200 mAh/g+caalyst) bio MnCo 2 0 4 nanowires (pink triangle) and bio CoMn 0 2 4 nanowires (green dot) tested at 1 A/ge (100 mA/gcaalyst) under I atm of 02. The electrodes were formed with 8 wt % of carbon and 72 wt % of catalyst materials. 105 4.6 References 1. Black R, Adams B, Nazar LF. Non-Aqueous and Hybrid Li-02 Batteries. Adv Energy Mater 2, 801-815 (2012). 2. Bruce PG, Freunberger SA, Hardwick LJ, Tarascon JM. Li-O-2 and Li-S batteries with high energy storage. Nat Mater 11, 19-29 (2012). 3. Chen YH, Freunberger SA, Peng ZQ, Fontaine 0, Bruce PG. Charging a Li-O-2 battery using a redox mediator. Nat Chem 5, 489-494 (2013). 4. Zhang T, Zhou HS. A reversible long-life lithium-air battery in ambient air. Nat Commun 4, (2013). 5. Lee JH, Black R, Popov G, Pomerantseva E, Nan FH, Botton GA, et al. The role of vacancies and defects in NaO.44MnO2 nanowire catalysts for lithium-oxygen batteries. Energ Environ Sci 5, 9558-9565 (2012). 6. Xu JJ, Xu D, Wang ZL, Wang HG, Zhang LL, Zhang XB. Synthesis of Perovskite-Based Porous LaO.75Sr0.25MnO3 Nanotubes as a Highly Efficient Electrocatalyst for Rechargeable LithiumOxygen Batteries. Angew Chem Int Edit 52, 3887-3890 (2013). 7. Ottakam Thotiyl MM, Freunberger SA, Peng Z, Chen Y, Liu Z, Bruce PG. A stable cathode for the aprotic Li-02 battery. Nat Mater 12, 1050-1056 (2013). 8. Jung HG, Jeong YS, Park JB, Sun YK, Scrosati B, Lee YJ. Ruthenium-Based Electrocatalysts Supported on Reduced Graphene Oxide for Lithium-Air Batteries. Acs Nano 7, 3532-3539 (2013). 9. Yamasaki Y, Miyasaka S, Kaneko Y, He JP, Arima T, Tokura Y. Magnetic reversal of the ferroelectric polarization in a multiferroic spinel oxide (vol 96, pg 207204, 2006). Phys Rev Lett 96, (2006). 10. Cheng FY, Shen JA, Peng B, Pan YD, Tao ZL, Chen J. Rapid room-temperature synthesis of nanocrystalline spinels as oxygen reduction and evolution electrocatalysts. Nat Chem 3, 79-84 (2011). IL. Courtel FM, Duncan H, Abu-Lebdeh Y, Davidson IJ. High capacity anode materials for Li-ion batteries based on spinel metal oxides AMn(2)O(4) (A = Co, Ni, and Zn). JMater Chem 21, 10206-10218 (201 1). 12. Nam KT, Kim DW, Yoo PJ, Chiang CY, Meethong N, Hammond PT, et al. Virusenabled synthesis and assembly of nanowires for lithium ion battery electrodes. Science 312, 885-888 (2006). 13. Nissinen T, Valo T, Gasik M, Rantanen J, Lampinen M. Microwave synthesis of catalyst spinel MnCo204 for alkaline fuel cell. JPowerSources 106, 109-115 (2002). 106 14. Rios E, Gautier JL, Poillerat G, Chartier P. Mixed valency spinel oxides of transition metals and electrocatalysis: case of the MnxCo3-x04 system. ElectrochimActa 44, 149 1- 1497 (1998). 15. Wang L, Zhao X, Lu YH, Xu MW, Zhang DW, Ruoff RS, et al. CoMn2O4 Spinel Nanoparticles Grown on Graphene as Bifunctional Catalyst for Lithium-Air Batteries. J Electrochem Soc 158, A 1379-A 1382 (2011 ). 16. Freunberger SA, Chen YH, Peng ZQ, Griffin JM, Hardwick L, Barde F, et al. Reactions in the Rechargeable Lithium-O-2 Battery with Alkyl Carbonate Electrolytes. JAm Chem Soc 133, 8040-8047 (2011). 17. Lavela P, Tirado JL, Vidal-Abarca C. Sol-gel preparation of cobalt manganese mixed oxides for their use as electrode materials in lithium cells. Electrochim Acta 52, 7986- 7995 (2007). 18. Liu L, Yang YZ. Shape-controlled synthesis of Mn-Co complex oxide nanostructures via a polyol-based precursor route and their catalytic properties. Superlattice Microst 54, 2638 (2013). 19. Li JF, Xiong SL, Li XW, Qian YT. A facile route to synthesize multiporous MnCo204 and CoMn204 spinel quasi-hollow spheres with improved lithium storage properties. Nanoscale 5,2045-2054 (2013). 20. Rosant C, Avalle B, Larcher D, Dupont L, Friboulet A, Tarascon JM. Biosynthesis of Co304 electrode materials by peptide and phage engineering: comprehension and future. Energ Environ Sci 5, 9936-9943 (2012). 21. Nuraje N, Dang XN, Qi JF, Allen MA, Lei Y, Belcher AM. Biotemplated Synthesis of Perovskite Nanomaterials for Solar Energy Conversion. Adv Mater 24, 2885-2889 (2012). 22. D Oh JQ, YC Lu, Y Zhang, Y Shao-Horn, A M Belcher. Biologically enhanced cathode design for improved capacity and cycle life for lithium-oxygen batteries. Nat Commun 4, 2756 (2013). 107 Chapter 5. Ongoing work: Probing graphene defects with phage display 108 5.1 Introduction Graphene has showed its great potential with its excellent electrical', quantum mechanical2 and mechanical3' 4 properties. In particular, mass production of defect free graphene sheets in large scale (~M2) is crucial in flexible display, solar panel and electronic device fabrications.5 However, currently available methods to detect the graphene defect are limited to scanning techniques, such as Atomic force microscope (AFM), 6 Transmission electron microscopy (TEM) 7 and Raman spectroscopy 8 . Thus, an easy and single-step toolkit to screen the impurity of graphene needs to be developed to expand its application into industrial scale. The material specific affinity of protein assembly can be used to detect the impurity of products,9 for example, graphene defects. Here, a M 13 virus based graphene defect detector is proposed by implementing the dual functionality in a single biomolecule. As each type of M13 virus coat protein can be separately designed with a different functionality, the detector and signal producer can be equipped into p3 and p8 of M 13 virus (Fig. 5.1). By selecting a specific peptide sequence that favorably binds to graphene defects from M13 virus p3 library, this clone can be used to detect nanoscale defect on one layer of graphene sheet. Since the perfect graphene sheet exhibits stronger hydrophobicity than defect region (e.g. -COOH, -OH), we expect the interaction mechanism between graphene defect and peptide sequence can be easily used to differentiate the defect region. In addition, once the virus attached to the imperfect carbon bonds, the position of virus can be easily detected by applying fluorescence molecules on the body of M13 virus. This would help to detect the graphene defects with a single optical image of the products rather than using time-consuming analysis techniques. 109 5.2 Experimental 5.2.1 Selecting the graphene flake-binding genetic sequence by bio-panning method The graphene flake sample with an area of 0.5x0.5 cm 2 was prepared on a cupper foil. This sample was held on the 24-well plate with double sticky tapes and washed with I ml of TBS (IX) solution. The blocking buffer was added to completely cover (1 ml) the sample area and incubated for 1 hr with gentle rocking at room temperature. After the incubation, the sample was washed 6 times rapidly with TBST (0.1 % v/v, 300 [d). From the library stock solution (NEB, Ph.D.TM -C7C library), 2x10" phage was diluted with 390 [d of TBST (0.1 % v/v) and incubated with the graphene flake sample for 1 hr with gentle rocking at room temperature. Unbound phage was discarded after the incubation and the sample was washed with I ml of TBST (0.1 % v/v, from the second round, increase to 0.2 % v/v) with 10 times. The elution buffer (I ml of 0.2 M Glycine-H Cl, pH 2.2, 1 mg/ml of BSA) was added for 20 min with gentle rocking at room temperature and collected for neutralization with 150 [l of 1 M Tris-HCI (pH 9.1). For E.coli elution, 100 [l of mid-log culture was incubated for 5 min and directly followed for titrations. From each eluted phage solution (Glycine buffer and E.coli elution), 10 [d of virus was collected and amplified with 20 ml scale. In each round, the number of phage in library was fixed to 2x10". After the amplification, the phage solution was titered using agar plates containing Xgal/isopropyl-#f-D- I -thiogalactopyranoside (IPTG)/tetracycline. The DNA from each plaque was extracted using QlAprep* Miniprep Kit (Qiagen) and sequenced in the Biopolymers laboratory at MIT. 110 5.3 Results and discussion To select the genetic sequence having affinity towards graphene defects from Ph.D.TM - C7C library, graphene flakes were grown on Cu foil by a low pressure chemical vapor deposition (CVD) method (Fig. 5.2). The synthetic condition was adjusted to form graphene flakes rather than the perfect graphene layer because the flake has a high concentration of graphene edges or broken bonds. These flakes can be easily transferred to Si wafer so that we can disregard the interaction between the protein assembly and substrate materials. These graphene flakes samples with two different substrates (Cu, Si) will be applied alternatively for each round to minimize the substrate-mediated binding. The first round of bio-panning with C7C library was conducted against Cu-graphene flakes. To collect the bound phages, the acid elution was first applied and the E.coli solution was further added afterwards to harvest remaining phages. These phages were DNA sequenced by selecting the random plaques after titrations. The sequence results of phages eluted by E.coli solution and acid are included in Fig. 5.3. We observed the high frequency of Histidine (H), which functioned as the hydrophobic-hydrophobic interaction moiety with carbon-based materials,' 0 among the selected phages. The diversity of library was maintained after the first round as can be seen from the various combinations of sequences in Fig. 5.3. On the second round, the amplified library from the first round was added with the fixed number of phages. Unbound phages were washed with more harsh conditions (TBST, 0.2 % v/v) for the second round. However, the sequencing result after the second round of bio-panning converges into few clones that dominate in the population. This can be due to the difference of phage growth speed during the amplification after the selection. Thus, the amplification duration can be adjusted to the point where the library diversity maintains. 111 5.4 Figures M13 virus p8: Signal producer (Dye conjugated) p3: Graphene defect detector Figure 5.1. Scheme of M13 virus-based graphene defects detector. Ml13 virus based graphene defects detector (left) with dual functionality (p 8 : signal producer, p 3 : graphene defect detector) and the operation mechanism of this biosensor (right). 112 Figure 5.2. Scanning electron microscope (SEM) image of graphene flakes. The graphene flakes were grown with Cu-mediated low pressure CVD method by Mr. Yong Cheol Shin. The image credit goes to Mr. Shin. 113 1 st Round Gly C C G V S L A G C C C C C C C C C T L G L Q K R T F T K M A V P R E P A Q R T T L A I G N Q P L S G N K T S G D N A E A F I p A Q L G N K M A G T F P N S D S A D E A C K S T E T A S C N T G S p Y E C E R F N Q G L C N E p K p R L C C C T G p A R R 2"d Round E.coli Figure 5.3. p3 sequences of M13 virus clones selected from bio-panning against graphene flakes. The sequencing result after the first and second round of bio-panning against graphene flakes on Cu foil. The left column indicates the elution method. Each sequence contains two Cysteines in the beginning and at the end that form the ring in p3. 114 5.5 References 1. Geim AK, Novoselov KS. The rise of graphene. Nat Mater 6, 183-191 (2007). 2. Novoselov KS, Geim AK, Morozov SV, Jiang D, Katsnelson MI, Grigorieva IV, et al. Two-dimensional gas of massless Dirac fermions in graphene. Nature 438, 197-200 (2005). 3. Lee C, Wei XD, Kysar JW, Hone J. Measurement of the elastic properties and intrinsic strength of monolayer graphene. Science 321, 385-388 (2008). 4. Bunch JS, Verbridge SS, Alden JS, van der Zande AM, Parpia JM, Craighead HG, et al. Impermeable atomic membranes from graphene sheets. Nano Lett 8, 2458-2462 (2008). 5. Bae S, Kim H, Lee Y, Xu XF, Park JS, Zheng Y, et al. Roll-to-roll production of 30-inch graphene films for transparent electrodes. Nat Nanotechnol 5, 574-578 (2010). 6. Paredes I, Villar-Rodil S, Solis-Fernandez P, Martinez-Alonso A, Tascon JMD. 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