Characterization and Identification of Bacteria in a Consortium Capable of Degrading Residual Jatropha Biodiesel Wastewater Oil


Armi R. Creencia1, Bernadette C. Mendoza2, Veronica P. Migo1,
and Rosario G. Monsalud1
1National Institute of Molecular Biology and Biotechnology (BIOTECH),
University of the Philippines Los Baños, College, Laguna
1Institute of Biological Sciences (IBS),
University of the Philippines Los Baños, College, Laguna
corresponding author: This email address is being protected from spambots. You need JavaScript enabled to view it.



A consortium comprising of three bacteria had been found to have a potential in degrading residual Jatropha oil biodiesel wastewater. Based on a previous study, this promising consortium can degrade up to 97% of residual oil under partially optimized conditions for 8 d. The three bacteria were subjected to phenotypic and genotypic characterizations. Specifically, cultural, morphological, physiological and biochemical testing, and Fatty Acid Methyl Ester (FAME) profiling were conducted. Ribosomal DNA (16S rDNA) sequence analysis was also done. Isolates BOcMFW-2 and BOcMJL-12 were Gram-negative and Gram-positive mangrove bacteria, respectively. On the other hand, Isolate BDF-2 was a Gram-positive bacterium recovered from Jatropha wastewater. These bacteria were identified as: Pseudoalteromonas sp. (BOcMFW-2), Arthrobacter sp. (BOcMJL-12), and Bacillus cereus (BDF-2). The bacterial consortium can be further developed as microbial treatment for biodiesel wastewater.



The use of biodiesel has become a global trend in addressing environmental pollution problems. With biodiesel, biological and public health risks can also be reduced. Biodiesel wastewater generally contains water, glycerol, soaps, methanol, base/acid catalyst and residual oils. Lipids (i.e., fats, oil and grease) present in the wastewater are difficult to treat since they are water-insoluble (Matsumiya et al. 2007). Accumulation of these lipid-rich wastes may cause destruction of animal breeding grounds, corrosion of sewer lines, clogging of drainages and production of foul odors (Suehara et al. 2005; Matsumiya et al. 2007). Mechanical, physical separations and chemical treatments were found to be impractical and expensive (Bhumibhamon & Phattayakorn 2003). . . . . . . . . . . . . . . . . .





BHUMIBHAMON O, PHATTAYAKORN K. 2003. Lipase-producing microorganisms for use in contaminated fat and oil kitchen wastewater Treatment. Kasetsart J: Nat Sci 37: 327-333.

CAMMAROTA MC, FREIRE DMG. 2006. A review on hydrolytic enzymes in the treatment of wastewater with high oil and grease content. Bioresource Tech. 97: 2195-2210.

GARRITY GM, BRENNER DJ, KRIEG NR, STALEY JT. 2005. Bergey’s Manual of Systematic Bacteriology. 2nd ed. Vol. 2 Part B. New York, USA: Springer. 1106p.

HABA E, BRESCO O, FERRER C, MARQUES A, BUSQUETS M, MANRESA A. 2000. Isolation of lipase-secreting bacteria by deploying used frying oil as selective substrate. Enzym Microb Tech 26: 40-44.

HASAN F, SHAH AA, HAMEED A. 2006. Industrial applications of microbial lipases. Enzym Microb Tech 39: 235-251.

JONES D, KEDDIE R. 2006. The Genus Arthrobacter. In: Dworkin, Falkow MS, Rosenberg E, Schleifer K, Stackebrandt E, eds. The Prokaryotes. 3rd ed. Vol. 3. New York, USA: Springer. p. 945-960.

KUMAR S, TAMURA K, NEI M. 2004. MEGA3: Integrated software for Molecular Evolutionary Genetics Analysis and Sequence Alignment. Briefings in Bioinformatics 5: 150-163.

KUNITSKY C, OSTERHOUT G, SASSER M. 2000. Identification of microorganisms using fatty acid methyl ester (FAME) analysis and the MIDI Sherlock Microbial Identification System. Encyclopedia of Rapid Microbiological Methods. Newark, USA: MIDI, Inc. p. 1-17.

MADIGAN MT, MARTINKO JM, DUNLAP PV, CLARK DP. 2008. Brock Biology of Microorganisms. 12th ed. New Jersey, USA: Pearson Education, Inc. 341p.

MALIK KA. 1991. Maintenance of Microorganisms by Simple Methods. In: Kirsop, B.E. and A. Doyle. Maintenance of Microorganisms and Cultured Cells. 2nd ed. London: Academic Press, Inc. p. 121-132.

MATSUMIYA Y, WAKITA D, KIMURA A, SANPA S, KUBO M. 2007. Isolation and characterization of a lipid-degrading bacterium and its application to lipid-containing wastewater treatment. J Biosci Bioeng 103(4): 325-330.

MONGKOLTHANARUK W, DHARMSTHITI S. 2002. Biodegradation of lipid-rich wastewater by a mixed bacterial consortium. International Biodeterioration and Biodegradation 50(2): 101-105.

MONSALUD RG, ZULAYBAR TO, PAPA IA, CLAVILLAS JDLS. 2006. Isolation, Preservation and Identification of Microorganisms with Degradative, and Antimicrobial Potential from Mangrove Ecosystem in Mindoro. Project Terminal Report. Commission on Higher Education-Zonal Research Center (CHEDZRC) for Region IV.

RAYMUNDO AK, ZAMORA AF, DALMACIO IF. 1991. Manual on Microbiological Techniques. Technology and Livelihood Resource Center. University of the Philippines Los Baños: p. 17-31.

RECKNAGEL RD, BORMANN EJ, FRIEDRICH W. 1988. Improvement of medium composition by random balance designs. J Basic Microbiol 28: 659-666.

SUEHARA K, KAWAMOTO Y, FUJII E, KOHDA J, NAKANO Y, YANO T. 2005. Biological treatment of wastewater discharged from biodiesel fuel production plant with alkali-catalyzed transesterification. J. Bioscience and Bioeng 100(4): 437- 442.

TANO-DEBRAH K, FUKUYAMA S, OTONARI N, TANIGUCHI F, OGURA M. 1999. An inoculum for the aerobic treatment of wastewaters with high concentration of fats and oils. Bioresour Technol 69: 133-139. 

WILSON K. 1994. Preparation of genomic DNA from Bacteria. In: Ausubel FM, Brent R, Kingston RE, Moore D, Seidman JG, Smith JA, Struhl K. eds. Current Protocols in Molecular Biology. Vol. 1. USA: John Wiley and Sons, Inc. p. 241-242.