ISOLATION, PURIFICATION, AND CHARACTERIZATION OF LIPASE FROM BACILLUS SP. FROM KITCHEN GREASE

Jaiganesh R; Jaganathan Mk

doi:10.22159/ajpcr.2018.v11i6.24955

Authors

Jaiganesh R Department of Biotechnology, School of Bioengineering, SRM Institute of Science and Technology, Chennai, Tamil Nadu, India.
Jaganathan Mk Department of Biotechnology, School of Bioengineering, SRM Institute of Science and Technology, Chennai, Tamil Nadu, India.

DOI:

https://doi.org/10.22159/ajpcr.2018.v11i6.24955

Keywords:

Lipase, Esterase, Bacillus sp, Purification, Biomass

Abstract

Objective: The objective of this work was to isolation, purification and characterization of solvent tolerant lipase from Bacillus sp. The objective of this work was to isolation, purification and characterization of solvent tolerant lipase from Bacillus sp. from kitchen grease for a variety of applications including organic synthetic reactions and preparation of enantiomerically pure pharmaceuticals.

Methods: Lipase producing isolates were screened from kitchen grease on a selective medium rhodamine B olive oil agar, and tributyrin agar was used to screen the lipase and esterase producing an organism, respectively. The isolate identified using 16S rDNA sequencing method and enzyme activity was quantitatively assayed. Lipase production was characterized in different conditions.

Results: The isolate showed highest lipase activity was which later was identified as Bacillus sp. using 16S rDNA sequencing method. The lipase was purified using ammonium sulfate precipitation. The isolate showed excellent tolerance to methanol, ethanol, acetonitrile, and moderate tolerance to butanol. The increased biomass concentration, maximum production, and activity were achieved at 37Â°C in 24 h incubation, then gradual reduction in production was observed. The maximum activity of lipase enzyme was obtained at pH between 6 and 9.

Conclusion: The isolate produce solvent tolerance lipase enzyme and it can be a promising candidate of solvent tolerance lipase enzyme for variety of industrial applications.

Downloads

Download data is not yet available.

Author Biographies

Jaiganesh R, Department of Biotechnology, School of Bioengineering, SRM Institute of Science and Technology, Chennai, Tamil Nadu, India.

Associate Professor

Department of Biotechnology

School of Bioengineering

Jaganathan Mk, Department of Biotechnology, School of Bioengineering, SRM Institute of Science and Technology, Chennai, Tamil Nadu, India.

Assistant Professor

Department of Biotechnology

School of Bioengineering

References

Sakai Y, Hayatsu M, Hayano K. Use of tween 20 as a substrate for assay of lipase activity in soils. Soil Sci Plant Nutr 2002;48:729-34.

Jaeger KE, Dijkstra BW, Reetz MT. Bacterial biocatalysts: Molecular biology, three-dimensional structures, and biotechnological applications of lipases. Annu Rev Microbiol 1999;53:315-51.

Benjamin S, Pandey A. Mixed-solid substrate fermentation. A novel process for enhanced lipase production by Candida rugosa. Acta Biotechnol 1998;18:315-24.

Vakhlu J, Kour A. Yeast lipases: Enzyme purification, biochemical properties and gene cloning. Electron J Biotechnol 2006;9:69-85.

Saxena RK, Ghosh PK, Gupta R, Davidson WS, Bradoo S, Gulati R. Microbial lipases: Potential biocatalysts for the future industry. Curr Sci 1999;77:101-15.

Hasan F, Shah AA, Hameed A. Industrial applications of microbial lipases. Enzyme Microb Technol 2006;39:235-51.

Arpigny JL, Jaeger KE. Bacterial lipolytic enzymes: Classification and properties. Biochem J 1999;343 Pt 1:177-83.

Aravindan R, Anbumathi P, Viruthagiri T. Lipase applications in food industry. Indian J Biotechnol 2007;6:141-58.

Gupta N, Mehra G, Gupta R. A glycerol-inducible thermostable lipase from Bacillus sp.: Medium optimization by a plackettâ€“burman design and by response surface methodology. Can J Microbiol 2004;50:361-8.

Park H, Lee KS, Chi YM, Jeong SW. Effects of methanol on the catalytic properties of porcine pancreatic lipase. J Microbiol Biotechnol 2005;15:296-1.

Thota P, Bhogavalli PK, Vallem PR, Venkateswar S. Studies on optimization of extracellular lipase from potential fungal strain (s) isolated from oil contaminated soil. J Microbiol Biotechnol Res 2012;2:418-25.

Jain MK. Methods in enzymology volume 284. Lipases: Part A, biotechnology edited by B. Rubin (Lipomed) and E. A. Dennis (university of Californiaâˆ’San Diego). Academic Press: San Diego. 1997. Xxxi. J Am Chem Soc 1998;120:5354.

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S, et al. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 2011;28:2731-9.

Winkler UK, Stuckmann M. Glycogen, hyaluronate, and some other polysaccharides greatly enhance the formation of exolipase by Serratia marcescens. J Bacteriol 1979;138:663-70.

Green AA, Hughes WL. [10] Protein fractionation on the basis of solubility in aqueous solutions of salts and organic solvents. Methods Enzymol 1955;1:67-90.

Shu CH, Xu CJ, Lin GC. Purification and partial characterization of a lipase from Antrodia cinnamomea. Process Biochem 2006;41:734-38.

Henriette C, Zinebi S, Aumaitre MF, Petitdemange E, Petitdemange H. Protease and lipase production by a strain of Serratia marcescens (532 S). J Ind Microbiol 1993;12:129-35.

Kanimozhi1 K, Devairrakam EJ, Jegadeeshkumar D. Production and optimization of lipase from Bacillus subtilis. Int J 2011;2:6-10.

Abigor RD, Uadia PO, Foglia TA, Haas MJ, Scott K, and Savary BJ. Partial purification and properties of lipase from germinating seeds of Jatropha curcas L., J. Am. Oil Chem. Soc 2002;79(11):1123-126.

Ogino H, Miyamoto K, Ishikawa H. Organic-solvent-tolerant bacterium which secretes organic-solvent-stable lipolytic enzyme. Appl Environ Microbiol 1994;60:3884-6.

Ogino H, Miyamoto K, Yasuda M, Ishimi K, Ishikawa H. Growth of organic solvent-tolerant Pseudomonas aeruginosa LST-03 in the presence of various organic solvents and production of lipolytic enzyme in the presence of cyclohexane. Biochem Eng J 1999;4:1-6.

Kumar S, Ramesh R, Bhosle NB, Sardesai S, Sheshshayee MS. Natural isotopic composition of nitrogen in suspended particulate matter in the bay of bengal. Biogeosciences 2004;1:63-70.

Abdou AM. Purification and partial characterization of Psychrotrophic serratia marcescens lipase. J Dairy Sci 2003;86:127-32.

Joseph B, Upadhyaya S, Ramteke P. Production of cold-active bacterial lipases through semisolid state fermentation using oil cakes. Enzyme Res 2011;2011:796407.

Mohan TS, Palavesam A, Immanvel G. Isolation and characterization of lipase-producing Bacillus strains from oil mill waste. Afr J Biotechnol 2008;7:2728-35.

Rodriguez JA, Mateos JC, Nungaray J, Gonzalez V, Bhagnagar T, Roussos S, et al. Improving lipase production by nutrient source modification using Rhizopus homothallicus cultured in solid state fermentation. Process Biochem. 2006;41(11):2264-2269.

Kumar KD, Usha KY, Satyanarayana SV, Sailaja V. Characterization of partially purified lipase from Saccharomyces cerevisiae. Int J Pharm Pharm Sci 2014;6:514-17.

Suji CM, Priya RS, Sivaraj R. Optimization of lipase production from different agroindustrial wastes by marine actinomycetes. Int J Pharm Pharm Sci 2014;6:292-4.

Veerapagu M, Narayanan AS, Ponmurugan K, Jeya KR. Screening selection identification production and optimization bacterial lipase from oil spilled soil. Asian J Pharm Clin Res 2013;6 Suppl 3:62-7.

Lomthaisong K, Buranarom Z, Niamsup Z. Investigation of isolated lipase producing bacteria from oil-contaminated soil with proteomic analysis of its proteins responsive to lipase inducer. J Biol Sci 2012;12:161-7.