Sains Malaysiana 46(1)(2017): 43–50

http://dx.doi.org/10.17576/jsm-2017-4601-06

Biodegradation of Tapis Crude Oil Using Consortium of Bacteria and Fungi: Optimization of Crude Oil Concentration and Duration of Incubation by Response Surface Methodology

(Biodegradasi Minyak Mentah Tapis Menggunakan Konsortium Bakteria dan Kulat: Pengoptimuman Kepekatan Minyak Mentah dan Tempoh Pengeraman dengan Kaedah Gerak Balas Permukaan)

AINON HAMZAH*, VIDYAH MANIKAN & NUR AIN FATHIHA ABD AZIZ

School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 UKM, Bangi, Selangor Darul Ehsan, Malaysia

Received: 2 April 2015/Accepted: 14 April 2016

ABSTRACT

Response surface analysis was conducted to optimize the concentrations of Tapis crude oil and duration of incubation in order to achieve optimal microbial growth and crude oil biodegradation. Central Composite Rotatable Design (CCRD) was employed, where the design contained 13 experimental runs with different combinations of incubation time and crude oil concentration. The cultures containing mineral salt medium (MSM) with varying crude oil concentrations were incubated at 30°C, pH6.5 with 150 rpm agitation for 120-336 h. The inoculum contained a consortium of previously identified as oil degrading bacteria and fungi, namely Pseudomonas aeruginosa UKMP-14T, Acinetobacter baumannii UKMP-12T and Trichoderma sp. UKMP-1M and UKMP-2M. The analysis showed the duration of incubation plays a significant role (p<0.05) in affecting the bacterial growth and percentage of total petroleum hydrocarbon (TPH) biodegradation, meanwhile concentrations of Tapis crude oil has insignificant effect on the responses. Interaction of the two variables was found to be significant in affecting all the three responses, namely bacterial population, fungal biomass and percentage of TPH biodegradation. It was predicted through the CCRD that the percentage of biodegradation can be optimized to reach 86% on the 270th h when 5% (v/v) crude oil was used. This predicted value was verified to be achievable and reproducible through validation experiments.

Keywords: Biodegradation; crude oil; response surface methodology

ABSTRAK

Analisis gerak balas permukaan digunakan untuk mengoptimumkan kepekatan minyak mentah Tapis (yang ditemui dalam perairan Malaysia) dan tempoh pengeraman untuk mencapai pertumbuhan mikrob dan biodegradasi minyak mentah yang optimum. Reka Bentuk Gubahan Memusat Berputar (CCRD) telah digunakan, dengan 13 larian reka bentuk eksperimen kombinasi kepekatan minyak mentah dan tempoh pengeraman yang berbeza. Kultur yang ditumbuhkan dalam medium garam mineral (MSM) dengan kepekatan minyak mentah yang berbeza dieram pada suhu 30°C dan pH6.5 dengan kadar goncangan 150 psm bagi tempoh eraman 120-336 jam. Inokulum yang digunakan mengandungi konsortium bakteria dan kulat pengurai minyak yang telah dikenal pasti dalam kajian sebelumnya, iaitu Pseudomonas aeruginosa UKMP-14T, Acinetobacter baumannii UKMP-12T dan Trichoderma sp. UKMP-1M dan UKMP-2M. Hasil analisis menunjukkan tempoh masa eraman memainkan peranan yang signifikan (p<0.05) dalam mempengaruhi pertumbuhan bakteria dan peratus biodegradasi, manakala kepekatan minyak mentah Tapis tidak memainkan peranan yang signifikan. Interaksi antara dua pemboleh ubah juga didapati mempunyai kesan yang signifikan ke atas ketiga-tiga gerak balas yang dikaji, iaitu populasi bakteria, biojisim kulat dan peratus biodegradasi minyak mentah. Berdasarkan perisian CCRD, peratus biodegradasi yang optimum, iaitu 86% boleh dicapai pada jam yang ke-270 apabila 5% (v/v) minyak mentah digunakan. Jangkaan kombinasi optimum ini telah diuji dan disahkan melalui eksperimen.

Kata kunci: Analisis gerak balas permukaan; biodegradasi; minyak mentah

REFERENCES

Adebusoye, S.A., Ilori, M.O., Amund, O.O., Teniola, O.D. & Olatope, S.O. 2007. Microbial degradation of petroleum hydrocarbons in a polluted tropical stream. World Journal of Microbiology and Biotechnology 23(8): 1149-1159.

Agarry, S.E., Owabor, C.N. & Yusuf, R.O. 2012. Enhanced bioremediation of soil artificially contaminated with kerosene: Optimization of biostimulation agents through statistical experimental design. Journal of Petroleum & Environmental Biotechnology 3(3): 2-8.

Ainon Hamzah, Chia-Wei Phan, Pek-Hoon Yong & Nurul Hayati Mohd Ridzuan. 2014. Oil palm empty fruit bunch and sugarcane bagasse enhance the bioremediation of soil artificially polluted by crude oil. Soil and Sediment Contamination: An International Journal 23(7): 751-762.

Ainon Hamzah, Chia-Wei Phan, Nur Faizah Abu Bakar & Kok- Kee Wong. 2013. Biodegradation of crude oil by constructed bacterial consortia and the constituent single bacteria isolated from Malaysia. Bioremediation Journal 17(1): 1-10.

Ainon Hamzah, Mazni Abu Zarin, Aidil Abdul Hamid, Othman Omar & Sahidan Senafi. 2012. Optimal physical and nutrient parameters for growth of Trichoderma virens UKMP-1M for heavy crude oil degradation. Sains Malaysiana 41(1): 71-79.

Ainon Hamzah, Amir Rabu, Raja Farzarul Hanim Raja Azmy & Noor Aini Yussoff. 2010. Isolation and characterization of bacteria degrading Sumandak and South Angsi Oils. Sains Malaysiana 39(2):161-168.

Bao, M.T., Wang, L.N., Sun, P.Y., Cao, L.X., Zou, J. & Li, Y.M. 2012. Biodegradation of crude oil using an efficient microbial consortium in a simulated marine environment. Marine Pollution Bulletin 64(6): 1177-1185.

Bento, F.M., Camargo, F.A.O., Okeke, B.C. & Frankenberger, W.T. 2005. Comparative biodegradation of soil contaminated with diesel oil by natural attenuation, biostimulation and bioaugmentation. Resource Technology 96: 1049-1055.

Brooijmans, R.J.W., Pastink, M.I. & Siezen, R.J. 2009. Hydrocarbon-degrading bacteria: The oil-spill clean-up crew. Microbial Biotechnology 2(6): 587-594.

Chaillan, F., Le Flèche, A., Bury, E., Phantavong, Y.H., Grimont, P., Saliot, A. & Oudot, J. 2004. Identification and biodegradation potential of tropical aerobic hydrocarbon-degrading microorganisms. Research in Microbiology 155(7): 587-595.

Changyi, Z., Qingyuan, L. & Guocheng, S. 2009. Isolation, characterization and study on properties of a strain for degradation of offshore petroleum hydrocarbons. Journal of Jimei University 14(1): 24-28.

Das, N. & Chandran, P. 2011. Microbial degradation of petroleum hydrocarbon contaminants: An overview. Biotechnology Research International 2011: 1-13.

Ghazali, F.M., Rahman, R.N.Z., Salleh, A.B. & Basri, M. 2004. Biodegradation of hydrocarbons in soil by microbial consortium. International Biodeterioration & Biodegradation 54: 61-67.

Haines, J.R., Koran, K.M., Holder, E.L. & Venosa, A.D. 2003. Protocol for laboratory testing of crude-oil bioremediation products in freshwater conditions. Journal Indian Microbiology Biotechnology 30: 107-113.

Ijaha, U.J.J. & Antaib, S.P. 2013. Removal of Nigerian light crude oil in soil over 12-month period. International Biodeterioration Biodegradation 51: 93-99.

Mitsika, E.E., Christophoridis, C. & Fytianos, K. 2013. Fenton and Fenton-like oxidation of pesticide acetamiprid in water samples: Kinetic study of the degradation and optimization using response surface methodology. Chemosphere 93(9): 1818-1825.

Mohajeri, L., Aziz, H.A., Zahed, M.A., Mohajeri, S., Kutty, S.R.M. & Isa, M.H. 2011. Response surface analysis and modeling of n-alkanes removal through bioremediation of weathered crude oil. Water Science and Technology 63(4): 618-626.

Mohajeri, L., Aziz, H.A., Isa, M.H. & Zahed, M.A. 2010. A statistical experiment design approach for optimizing biodegradation of weathered crude oil in coastal sediments. Bioresource Technology 101(3): 893-900.

Moneke, A. & Nwangwu, C. 2011. Studies on the bioutilization of some petroleum hydrocarbons by single and mixed cultures of some bacterial species. African Journal of Microbiology Research 5(12): 1457-1466.

Muthuswamy, S., Binupriya, A.R., Baik, S. & Yun, S. 2008. Biodegradation of crude oil by individual bacterial strains and a mixed bacterial consortium isolated from hydrocarbon contaminated areas. Clean 36(1): 92-96.

Rahman, K.S.M., Thahira-Rahman, J., Lakshmanaperumalsamy, P. & Banat, I.M. 2002. Towards efficient crude oil degradation by a mixed bacterial consortium. Bioresource Technology 85: 257-261.

Sathishkumar, M., Binupriya, A.R., Baik, S. & Yun, S. 2008. Biodegradation of crude oil by individual bacterial strains and a mixed bacterial consortium isolated from hydrocarbon contaminated areas. Clean 36: 92-96.

Sharma, S.L. & Pant, A. 2001. Crude oil degradation by marine actinomycete Rhodococcus sp. Indian Journal of Marine Sciences 30: 146-150.

Sharma, A. & Rehman, M.B. 2009. Laboratory scale bioremediation of diesel hydrocarbon in soil by indigenous bacterial consortium. Indian Journal of Experimental Biology 47: 766-769.

Thenmozhi, R., Nagasthya, A. & Thajuddin, N. 2011. Studies on biodegradation of used engine oil by consortium cultures. Advances in Environmental Biology 5(6): 1051-1057.

Throne-Holst, M., Wentzel, A., Ellingsen, T.E., Kotlar, H.K. & Zotchev, S.B. 2007. Identification of novel genes involved in long-chain n-alkane degradation by Acinetobacter sp. strain DSM 17874. Applied and Environmental Microbiology 73(10): 3327-3332.

Zahed, M.A., Aziz, H.A., Isa, M.H. & Mohajeri, L. 2010. Enhancement biodegradation of n-alkanes from crude oil contaminated seawater. International Journal of Environmental Research 4(4): 655-664.

*Corresponding author; email: antara@ukm.edu.my