Biodegradation activities of
Phanerochaete chrysosporium
fungus and the bacteria
Pseudomonas putida and
Sphingomonas macrogoltabidus on
pretreated HDPE plastic
Anne Richards, Cara Broshkevitch, Ong Kim Yao,
and Poh Yong Rui
Background



HDPE plastic: used in food packaging, plastic bags, plastic
bottles, recycled plastic lumber, toiletry and liquid
containers, outdoor furniture, and piping
Takes over 100 years to degrade in a landfill-discarded
plastics filling up landfills (Sivan, 2011)
Detrimental environmental effects:



Water and air cannot enter soil – deplete underground water
resource
Disrupts degradation of other substances
In the ocean, plastic chokes and entangles marine life
Background continued

Other ways of disposal can not be used:


Burning plastics at high temperatures produces
toxic irritants that are human carcinogens
Past research: biodegradation with bacteria or
fungi, UV radiation, and thermal radiation have
been separately investigated

All three organisms have been shown to degrade
polyethylene plastic separately
Purpose
To determine the optimum treatment of
HDPE plastic for maximum biodegradation.
Academy of Science
Hypothesis:
The following factors affect
the rate of biodegradation of
HDPE plastic mass:

Ratios of different
microorganisms



P. Chrysosporium fungus
P. Putida bacteria
S. Macrogoltabidus bacteria
Hwa Chong Institution
Hypothesis:
The following factors affect
the rate of biodegradation of
HDPE plastic mass:


Environmental conditions for
biodegradation
Exposure time to UV
radiation
Variables:
Academy of Science


Independent variables
 Ratios of P.
chrysosporium fungus, P.
putida bacteria, and S.
macrogoltabidus bacteria
Dependent variable
 Percent change in dry
mass of the HDPE
plastic samples
 Amount of CO2 gas
present
Hwa Chong Institution

Independent variables



Environmental conditions for
biodegradation
Exposure time to UV radiation
Dependent variables




Percentage change in dry mass
of HDPE samples
Amount of CO2 gas present
Temperature of contents of flask
Cell density of bacterial cultures
Procedure
Academy of Science
Hwa Chong Institution
Culture Bacteria and Fungi
Culture Bacteria
Pre-treat Plastic
Prepare Soil
Conditions
Expose Plastic to Individual
Microorganisms
Pre-treat Plastic
Expose Plastic to different
Ratios of 3 Microorganisms
Expose Plastic to
Microorganisms
Measure Dependent
Variables
Measure Dependent
Variables
Microorganism Culture (AOS & HCI)
Bacteria will be cultured in
culture test tubes with
nutrient broth
Fungi will be cultured in
Petri dishes in potato
dextrose agar
Prepare Soil Conditions (HCI)
Erlenmeyer flask with
minimal media
Erlenmeyer flask with loamy soil
and distilled water
Pre-treat HDPE Plastic (AOS & HCI)
A
O
S
H
C
I
HDPE Plastic
Grocery Bags
HDPE Plastic
Grocery Bags
Cut into one
gram pieces
Cut into one
gram pieces
Exposed to
365nm UV
radiation for 96
hours
Exposed to
thermal radiation
in the oven at
115°C for 48
hours
Exposed to
365nm UV
radiation
for 72, 96,
120 hours
Exposed to
thermal radiation
in the oven at
115°C for 48
hours
Expose Plastic to Microorganisms (AOS & HCI)
Academy of Science
Expose plastic to:
 Individual
microorganisms




P. putida
S. macrogoltabidus
P. chrysosporium
Different ratios of
the three
microorganisms
Hwa Chong Institution
Expose plastic to:
 P. putida in:


Minimal Media
Loamy soil +
Distilled
water
Measure Dependent Variables (AOS & HCI)
Analytical Balance
CO2 Probe
Thermometer
Spectrophotometer
Will be used to measure:




Percentage change in dry mass of HDPE samples
Amount of CO2 gas present
Temperature of contents of flask
Cell density of bacterial cultures
A
O
S
H
C
I
Bibliography
Aamer Ali Shah (2007). Role of Microorganism in Biodegradation of Plastics. Retrieved October 30, 2011 from
http://eprints.hec.gov.pk/2361/1/2216.htm
Abraham, J., Nanda, S., & Sahu, S. (2010). Studies on the biodegradation of natural and synthetic polyethylene by
Pseudomonas spp. Journal of Applied Sciences and Environmental Management, 14.
Albano, C., Karam, A., Gonzalez, G., Dominguez, N., Sanchez,Y., Manzo, F. & Guzman-Garcia, C. (2005). Effect of gamma
irradiation on HDPE/HA (80:20) composites. Polymers for Advanced Technologies, 16, 283–285. Retrieved October
25, 2011 from http://onlinelibrary.wiley.com/doi/10.1002/pat.580/pdf
Ammala, A., Bateman, S., Dean, K., Petinakis, E., Sangwan, P., Wong, S.,Yuan, Q.,Yu, L., Patrick, C., & Leong, K.H. (2011). An
overview of degradable and biodegradable polyolefins. Progress in Polymer Science, 36, 1015-1049.
doi:10.1016/j.progpolymsci.2010.12.002
Anthony L. Andrady (1999). Environmental Degradation of Plastics under Land and Marine Exposure Conditions. Retrived
October 30, 2011 from http://www.5gyres.org/media/Environmental_Degradation%20of%20Plastics_by_Andrady.pdf
Arkatkar, A., Arutchelvi, J., Sudhakar, M., Bhaduri, S., Uppara, P.V., & Doble, M. (2009). Approaches to enhance the
biodegradation of polyolefins.The Open Environmental Engineering Journal, 2, 68-80.
Artham, T., & Doble, M. (2009). Biodegradation of physicochemically treated polycarbonate by fungi. Biomacromolecules,
11, 20-28. doi: 10.1021/bm 9008099
Arutchelvi, J., Sudhakar, M., Arkatkar, Ambika, Doble, Mukesh, Bhaduri, Sumit & Uppara, Parasu Veera (2008). Biodegradation
of polyethylene and polypropylene. Indian Journal of Biotechnology, 7, 9–22. Retrieved October 25, 2011 from
http://nopr.niscair.res.in/bitstream/123456789/7326/4/IJBT%207%281%29%209-22.pdf
CDC/Office of safety, health, and environment. (2010, November 10). Biosafety in microbiological and biomedical
laboratories (BMBL) 5th edition. Retrieved from http://www.cdc.gov/biosafety/publications/bmbl5/index.htm
Bibliography Continued…
Chowdhury,T., Ghosh, A., & Gupta, S. B. (2010). Isolation and selection of stress tolerant plastic loving bacteria isolates from
old plastic wastes. World Journal of Agricultural Sciences, 2, 138-140.
Chung, I.Y., Kim, E., Park, J.M., & Seo, S.W. (2008). A case of postoperative Sphingomonas paucimobilis endophthalmitis after
cataract extraction. Korean Journal of Ophthalmology, 22, 63-65. doi: 10.3341/kjo.2008.22.1.63
Gijsman, Pieter, Meijers, Guido & Vitarelli, Giacomo (1999). Cornparison of the UV-degradation chemistry of polypropylene,
polyethylene, polyamide 6 and polybutylene terephthalate. Polymer Degradation and Stability, 65, 433–441. Retrieved
October 25, 2011 from
http://cid.ispa.asso.fr/GEIDEFile/Degrad_0001.PDF?Archive=191929191910&File=Degrad+0001_PDF
He, Q., Liang, S., Wang,Y., Wei, C., & Wu, H. (2009). Degradation of o-chloronitrobenzene as the sole carbon and nitrogen
sources by Pseudomonas putida OCNB-1. Science Direct, 21, 89-95. doi: 10.1016/S1001-0742(09)60016-4
Huang,Yi-Li, Li, Qing-Biao, Deng, Xu, Lu,Ying-Hua, Liao, Xin-Kai, Hong, Ming-Yuan & Wang,Yan (2004). Aerobic and anaerobic
biodegradation of polyethylene glycols using sludge microbes. Process Biochemistry, 40, 207–211. Retrieved October 25,
2011 from http://envismadrasuniv.org/Biodegradation/pdf/Glycols%20using%20sludge%20microbes.pdf
Johnson, Kenneth E., Pometto, Anthony L. III & Nikolov, Zivko L. (1993). Degradation of Degradable Starch-Polyethylene Plastics
in a Compost Environment. American Society for Microbiology, 59, 1155–1161. Retrieved November 8, 2011 from
http://aem.asm.org/content/59/4/1155.full.pdf
Lanini, S., Houi, D., Aguilar, O. & Lefebvre, X. (2001).The role of aerobic activity on refuse temperature rise: II. Experimental and
numerical modelling. Waste Management & Research, 19, 58–69. Retrieved October 25, 2011 from
http://wmr.sagepub.com/content/19/1/58.full.pdf
Lucas, N., Bienaime, C., Belloy, C., Queneudec, M., Silvestre, F., & Nava-Saucedo, J. (2008). Polymer biodegradation:
Mechanisms and estimation techniques. Chemosphere, 73, 429-442. doi:10.1016/j.chemosphere.2008.06.064
Bibliography Continued…
Morancho, J.M., Ramis, X., Fernández, X., Cadenato, A., Salla, J.M., Vallés, A., Contat, L. & Ribes, A. (2006). Calorimetric and thermogravimetric
studies of UV-irradiated polypropylene/starch-based materials aged in soil. Polymer Degradation and Stability, 91, 44–51. Retrieved
November 8, 2011 from http://anvalllu.webs.upv.es/papers/2006_DSCTGAUVPPstarchsoil.pdf
Moriyoshi, K., Ohe, T., Ohmoto, T., Sakai, K., & Yamanaka, H. (2006). Biodegradation of Bisphenol A and related compounds by Sphingomonas sp.
Strain BP-7 isolated from seawater. Bioscience, Biotechnology, and Biochemistry, 71, 51-57.
Mostafa, H. M., Sourell, H. & Bockisch, F. J. (2010). The Mechanical Properties of Some Bioplastics Under Different Soil Types for Use as a
Biodegradable Drip Tubes. Agricultural Engineering International: the CIGR Ejournal, 12, 1–16. Retrieved November 7, 2011 from
http://www.cigrjournal.org/index.php/Ejounral/article/viewFile/1497/1270
Nanda, Sonil, Sahu, Smiti Snigdha & Abraham, Jayanthi (2010). Studies on the biodegradation of natural and synthetic polyethylene by
Pseudomonas spp. Journal of Applied Sciences & Environmental Management, 14, 57–60. Retrieved October 29, 2011 from
http://www.bioline.org.br/pdf?ja10027
Orhan, Yüksel, Hrenovićb, Jasna & Büyükgüngöra, Hanife (2004). Biodegradation of plastic compost bags under controlled soil conditions. Acta
Chimica Slovenica, 51, 579–588. Retrieved November 7, 2011 from http://acta.chem-soc.si/51/51-3-579.pdf
Shah, A., Hasan, F., Hameed, A., & Ahmed, S. (2008). Biological degradation of plastics: A comprehensive review. Biotechnology Advances, 26,
246-265. doi:10.1016/j.biotechadv.2007.12.005
Sharma, N., & Singh, B. (2008). Mechanistic implications of plastic degradation. Polymer Degradation and Stability, 93, 561-584.
doi:10.1016/j.polymolegradstab.2007.11.008
Sierra, Isabel, Valera, José Luis, Marina, M. Luisa & Laborda, Fernando (2003). Study of the biodegradation process of polychlorinated biphenyls in
liquid medium and soil by a new isolated aerobic bacterium (Janibacter sp.). Chemosphere, 53, 609–618. Retrieved November 7, 2011
from http://infolib.hua.edu.vn/Fulltext/ChuyenDe2009/CD240/60.pdf
Sivan, A. (2011). New perspectives in plastic biodegradation. Science Direct. doi: 10.1016/j.copbio.2011.01.013
Thraves, P., & Packer, R. (April 2009). Material safety data sheet. Retrieved from
http://www.hpacultures.org.uk/media/DF9/A0/Growing_Cultures_MSDS.pdf
Verma, Shefali (2002). Anaerobic Digestion of Biodegradable Organics in Municipal Solid Wastes. Retrieved October 25, 2011 from
http://www.seas.columbia.edu/earth/wtert/sofos/Verma_thesis.pdf