Wastes from Various Sectors of
Pakistan: Potential Raw Materials for
Biofuels & Biomaterials
Dr. Taous Khan
Department of Pharmacy,
COMSATS, Abbottabad, Pakistan
INTRODUCTION
Generation of wastes is increasing due to:
– growing urbanisation &
– changes in life style
~1.6 billion metric tons of solid waste is produced per year
A lot of money is used managing this waste
– Asian countries spent ~US$25 billion per year in early 1990s
– expected to rise to ~US$50 billion by 2025
Mahar et al., Proceedings of the International Conference on Sustainable Solid Waste Management,
5 -7 September 2007, Chennai, India. pp.34-41
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INTRODUCTION
Threat to our already degraded environment
Advances in biotechnology, genetics, chemistry & engineering
– new manufacturing concepts
– converting waste materials to valuable fuels & biomaterials
Kyungpook National University, Biochemical Engineering Lab
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Wastes from various sectors of Pakistan
Large amount of waste materials produced from various sectors
of Pakistan
– industrial sectors
– agricultural sectors
These materials are a source of:
– environmental pollution
– water pollution
– different diseases
COMSATS University, Abbottabad
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Wastes from various sectors of Pakistan
These materials are:
– rich in various elements including carbon & nitrogen
– can be used as substrate for microbial growth
– production of useful metabolic products
Thus, these waste materials can be a good source for
production of bioenergy & biomaterials
– their use can address several societal needs
– will lead to a new manufacturing paradigm
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Wastes from various sectors of Pakistan
In America
– 2500 MW electricity
– generated by waste-to-energy plants
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Physical Composition of Wastes
Three general categories of solid waste in Pakistan:
– biodegradable e.g. food & animal wastes, leaves, grass & wood
– non-biodegradable e.g., plastic, rubber, textile waste, metals, stones
– recyclable material e.g., paper, card board, and bones
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Physical Composition of Wastes
Figure. Physical composition of solid wastes in Pakistan (% Weight).
Mahar et al., Proceedings of the International Conference on Sustainable Solid
Waste Management, 5 -7 September 2007, Chennai, India. pp.34-41
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Wastes from various sectors of Pakistan
Household
Agriculture
Units
Sugar Mills
Wood
Processing
Industry
Hotels &
Restaurants
Waste
Materials
Leather
Industry
Beverage
Industry
Food
Industry
Meat
Industry
Figure. Various sources of waste materials from Pakistan.
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Potential Products from Wastes from various Sectors of Pakistan
Bioethanol
Bacterial
Polysacchari
des
Bacterial
cellulose
Biogas
Waste
Materials
Biologically
active
compounds
Biofertilizer
Enzymes
Antibiotics
Figure. Various products that can be produced from waste materials from Pakistan.
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A case study – Production of WSOS & BC from Wastes of Korean Breweries
WSOS & BC successfully produced from synthetic media
Expensive process, hindering their commercial applications
Alternate, cheaper culture media for production of these products
Waste from beer fermentation broth
– large-scale availability from Korean breweries
– contains carbon, nitrogen, sulfur & ethanol
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Composition of WBFB
Table. Detailed composition of the waste from beer fermentation broth.
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Importance of Oligosaccharides
Medical

Infectious & inflammatory diseases

Metabolic & cardiovascular disorders

Transplantation & neutralization of toxins

Cancer immunotherapy
Food

Pre-biotics

Bulking agents in diet foods
Agriculture

Activation of plant cell machinery

Fertilizers
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Bacterial Cellulose
Figure. Organic constituents of wood.
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Bacterial Cellulose
Pure, free from lignin, pectin & hemicellulose
High degree of crystallinity & water retention value
High tensile strength and moldability
Biodegradable (eco-friendly materials)
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Applications of Bacterial Cellulose
Figure. Important applications of bacterial cellulose.
http://www.rish.kyoto-u.ac.jp/houga/researches/2006m05a.jpg
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Gluconacetobacter hansenii PJK
Gluconacetobacter hansenii PJK
– Gram-negative bacteria
– originally isolated from rotten apples
Capable of producing
– water soluble oligosaccharides (WSOS)
– bacterial cellulose (BC)
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D-Glucose
T. Naritomi et al., J. Ferment. Bioeng. 85(1998) 598.
ATP
ADP
Glucose Kinase
Inhibition
S. Bielecki, et al., (2004) In: Biopolymers, A. Steinbuchel, ed., WILEY-VCH
Production of BC & WSOS by G. hansenii PJK
D-Glucose-6-Phosphate Glucose Phosphate Dehydrogenases
Pentose Phosphate
Pathway
Phosphoglutamase
ATP
T. Naritomi et al., J. Ferment. Bioeng. 85(1998) 598.
D-Glucose-1-Phosphate
Ethanol
Pyrophosphorylase
S. Bielecki, et al., (2004) In: Biopolymers, A. Steinbuchel, ed., WILEY-VCH
UDP-D-Glucose
2NAD
Cellulose synthase
Cellulose
UDP-D-glucose dehydrogenase
2NADH
UDP-D-Glucuronic Acid
I.W. Sutherland, Int. Dairy J., 11 (2001) 663-674
http://glucuronicacid.quickseek.com/
Glucuronide
UDP
D-Glucuronic Acid
NAD
Aldehyde reductase
NADH
Daly, A.K.; Mantle, T.J.T., Biochem. J. 205 (1982) 381-388
http://glucuronicacid.quickseek.com/
L-Gulonic Acid
Figure. Proposed biosynthetic pathway for the simultaneous production of BC & WSOS from glucose by G. hansenii PJK
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Pre-treatment of WBFB
Figure. Schematic representation for the processing of WBFB.
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Bioreactors Configuration
(a)
(b)
Figure. Schematic diagrams of jar fermenters used for the production of WSOS and BC by G.
hansenii PJK; 5 L jar fermenter (a) and 5 L jar fermenter equipped with a spin filter (b).
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Bioreactors Configuration
(a)
(b)
Figure 19. Selected pictures of the 2 L jar fermenter equipped with a spin filter; empty fermenter
(a) and with 1.6 L of culture broth.
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Measurement of WSOS, BC & Cells
Figure. Schematic representation for the measurement of WSOS, BC and Cells from the culture broth..
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Culture Conditions
Table. Detailed fermentation conditions used for the production of WSOS and BC by G. hansenii PJK.
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WSOS & BC Production using WFBF
Effect of dilution ratio
Figure. Production of WSOS & BC by G. hansenii PJK in baffled flasks shaken at 100 rpm at
30 oC for 7 days, using the WBFB diluted with distilled water, in various ratios.
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WSOS & BC Production using WFBF
Effect of impeller speed
Figure. Production of WSOS & BC by G. hansenii PJK in a 5 L jar fermenter (3 L working
volume), an aeration rate of 1 vvm, a pH of 4.5-5.5 and impeller speeds of 300 rpm.
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WSOS & BC Production using WFBF
Effect of impeller speed
Figure. Production of WSOS & BC by G. hansenii PJK in a 5 L jar fermenter (3 L working
volume), an aeration rate of 1 vvm, a pH of 4.5-5.5 and impeller speeds of 500 rpm.
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WSOS & BC Production using WFBF
Effect of impeller speed
Figure. Production of WSOS & BC by G. hansenii PJK in a 5 L jar fermenter (3 L working
volume), an aeration rate of 1 vvm, a pH of 4.5-5.5 and impeller speeds of 600 rpm.
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Overproduction of WSOS obtained in present study
Table. Comparison of production of WSOS obtained in present study with products of related chemical nature reported in literature.
[1]. Jung et al., Enzyme Microb. Technol., 37 (2005) 354.
[2]. Courtois et al., J. Carbohydr. Chem., 12 (1993) 448.
[3]. Michaud et al., Int. J. Biol. Macromol., 17 (1995) 369.
[4]. Michaud et al., Int. J. Biol. Macromol., 16 (1994) 301.
[5]. Heyraud et al., Carbohydr. Res., 240 (1993) 71.
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Structure of WSOS
Figure. Schematic representation of various techniques applied for structure determination of WSOS.
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Structure of WSOS
Structure of WSOS
Figure. Chemical structures and fragmentation scheme of the major WSOS produced from WBFB.
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Structure of WSOS
Chemical structure of WSOS :

Mixture of oligomers of glucuronic acid

Molecular weights less than 1000

α- Linked rather than β
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Potential applications of WSOS
WSOS produced from WBFB:

Glucuronan oligosaccharide
Glucuronan polysaccharides:

Carriers for various drugs

Solvents, stabilizers, binders and swelling agents
Medical properties of glucuronan polysaccharides

Bleeding stoppage during surgery

Prevention of post-surgical adhesions

Antibacterial activity

Bone regeneration
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Physical properties of WSOS
Figure. Schematic representation for the study of physical properties of WSOS.
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Physical properties of WSOS
Physical characteristics of WSOS from the defined medium:

Flake-type with porous surfaces

Free from adhered microbial cells contamination


Thermal stability better or comparable to the other microbial polysaccharides


No problems in its practical applications.
Can proceed for development into a valuable product
Good emulsifying activity with short term stability

Useful as an emulsifier in combination with a stabilizer

Alone in situations where only short-term emulsion stability is desired.
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Conclusion
Production of WSOS and BC by G. hansenii PJK from the WBFB :

WBFB – worthy substitute for the conventional defined medium

Better production yield of WSOS compared to synthetic medium
Physico-chemical properties of WSOS :

Few structural differences from the WSOS from a defined medium

The presence of O-acetyl and O-methyl groups

Lack of unsaturation in the terminal unit

Similar or even better thermal and emulsifying characteristics

Potential for development for commercial applications
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Our Current Research in Pakistan
Currently we are working to evaluate various waste
materials from different industrial & agricultural
sectors of Pakistan for the production of BC
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Publications
From this work
1.
Taous Khan, S.H. Hyun, J.K. Park, Production of glucuronan oligosaccharides using the
waste of beer fermentation broth as a basal medium, Enzyme and Microbial Technology,
42, 89-92 (2007).
2.
Taous Khan, J.K. Park, The structure and physical properties of glucuronic acid oligomers
produced by a Gluconacetobacter hansenii strain using the waste from beer fermentation
broth, Carbohydrate Polymers, 73(3), 438–445 (2008).
3.
J.H. Ha, O. Shehzad, S. Khan, S.Y. Lee, Taous Khan, J.K. Park, Production of bacterial
cellulose by a static cultivation using the waste from beer culture broth, Korean Journal of
Chemical Engineering, 25(4), 812–815 (2008).
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