Journal of Applied & Environmental Microbiology
ISSN (Print): 2373-6747 ISSN (Online): 2373-6712 Website: Editor-in-chief: Sankar Narayan Sinha
Open Access
Journal Browser
Journal of Applied & Environmental Microbiology. 2018, 6(1), 10-17
DOI: 10.12691/jaem-6-1-2
Open AccessArticle

Characterization of Lipase from Bacillus subtilis Isolated from Oil Contaminated Soil

Femi-Ola T. O.1, , Odeyemi A. T.1, Olaiya B. S.1 and Ojo O. O.1

1Department of Microbiology, Ekiti State University, Ado-Ekiti, Nigeria

Pub. Date: January 10, 2018

Cite this paper:
Femi-Ola T. O., Odeyemi A. T., Olaiya B. S. and Ojo O. O.. Characterization of Lipase from Bacillus subtilis Isolated from Oil Contaminated Soil. Journal of Applied & Environmental Microbiology. 2018; 6(1):10-17. doi: 10.12691/jaem-6-1-2


An extracellular lipase from Bacillus subtilis isolated from oil polluted soil was partially purified and characterized in this study. The enzyme was purified to 19.36 fold and the molecular weight was estimated. The effect of temperature, time, metal ion and pH was also determined. The molecular weight of the enzyme was estimated to be 48.63 kDa by SDS-PAGE. The optimum temperature was 60¡ãC, while the enzyme exhibited appreciable thermostability retaining 70% of activity at 70¡ãC for 1h. The lipase was most active in the pH range of 7-9 with an optimum activity at pH 8.0. The enzyme activity declined in the presence of Al3+ and Fe2+, while Na+ stimulated the activity. Olive oil was found to be the preferred substrate. The maximum velocity Vmax and Km of the lipase during the hydrolysis of olive oil were 39.45 ¦Ìmol/min/ml and 20.01 mM respectively.

Lipase olive oil Bacillus subtilis thermostability

Creative CommonsThis work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit


[1]  Reis, P.; Holmberg, K.; Watzke, H.; Leser, M. E. and Miller, R. (2008). Lipases at interfaces: A review. Advances in Colloid and Interface Science.
[2]  Park, H.; Lee, K.; Chi, Y. and Jeong, S. (2005). Effects of methanol on the catalytic properties of porcine pancreatic lipase. Journal of Microbiology and Biotechnology; 15(2), 296.
[3]  Gupta, N.; Shai, V. and Gupta, R. (2007). Alkaline lipase from a novel strain Burkholderiamultivorans: Statistical medium optimization and production in a bioreactor. Process Biochemistry; 42(2), 518.
[4]  Grbavcic, S. Z., Dimitrijevic-Brankovic, S. I., Bezbradica, D. I., Siler-Marinkovic, S. S. and Knezevi, Z. D. (2007). Effect of fermentation conditions on lipase production by Candidautilis. Journal of the Serbian Chemical Society; 72(8-9), 757.
[5]  Franken, L. P. G., Marcon, N. S., Treichel, H., Oliveira, D., Freire, D. M. G. and Dariva, C. (2009). Effect of treatment with compressed propane on lipases hydrolytic activity.Foodand Bioprocess Technology; 10:0087-0095.
[6]  Joseph, B., Ramteke, P. W. and Thomas, G. (2008). Cold active microbial lipases: Some hot issues and recent developments. Biotechnology Advances; 26: 457-470.
[7]  Schmid, R. and Verger, R. (1998). Lipases: Interfacial enzymes with attractive applications. Angew. Chemical International Edition; 37: 1608-1633.
[8]  Titball, R. W. (1998). Bacterial phospholipases. Symp. Ser. Soc. Applied Microbiology; 27:127S-137S.
[9]  Margesin, R.; Labb¨¦, D.; Schinner, F.; Greer, C. W. and Whyte, L. G. (2003). Characterization of hydrocarbon-degrading microbial populations in contaminated and pristine alpine soils. Applied Environmental Microbiology; 69: 3085-3092.
[10]  Jaeger, K.E. and Eggert, T. (2004). Enantioselective biocatalysis optimized bydirected evolution. Current Opinion Biotechnology; 15: 305-313.
[11]  Rosenau, F. and Jaeger, K. E. (2000). Bacterial lipases from Pseudomonas: Regulation of gene expression and mechanisms of secretion. Biochimie; 82:1023-1032.
[12]  Kahveci, D., Guo, Z., Ozcelik, B. and Xu, X. (2009). Lipase-catalyzedglycerolysis in ionic liquids directed towards diglyceride synthesis. Process Biochemistry; 44: 1358-1365.
[13]  Shah, S., Sharma, S. and Gupta, M. N. (2004).Biodiesel preparation by lipase catalysed transesterification of Jatropha oil. Energy Fuels; 18: 154-9.
[14]  Kumar, S., Kikon, K., Upadhyay, A., Kanwar, S. S.and Gupta, R. (2005). Production, purification and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3.Protein Expression and Purification; 41: 38-44.
[15]  Nawani, N., Khurana, J. and Kaur, J. (2006). A thermostable lipolytic enzyme from a thermophilic Bacillus species. Purification and characterization. Molecular Cellular Biochemistry; 290: 17-22.
[16]  Olusesan, A. T., Azura, L. K., Forghani, B., Abu Bakar, F., Mohamed, A. K., Radu, S., Abdul Manap, M. Y. and Saari, N. (2011). Purification, characterization and thermal inactivation kinetics of a non regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS N. Biotechnology.
[17]  Lima, V., Krieger, N., Mitchell, D., Baratti, J., De Filippis, I., and Fontana, J. (2004). Evaluation of the potential for use in biocatalysis of a lipase from a wild strain of Bacillusmegaterium. Journal of Molecular Catalyst and Biological Enzyme; 31: 53-61.
[18]  Kambourova, M., Kirilova, N.; Mandeva, R. and Derekova, A. (2003). Purification and properties of thermostable lipase from a thermophilic Bacillus stearothermophilus MC. Journal of Molecular Catalyst and Biological Enzymes; 7(22): 307-313.
[19]  Singh, M., Kumar, S., Neha, S. and Krishnan. K. (2010). Lipase Production by Bacillus subtilis OCR-4 in Solid State Fermentation Using Ground Nut Oil Cakes as Substrate. Current Research Journal of Biological Sciences 2(4): 241-245.
[20]  Prasad, M. P. and Sethi, R (2013). Comparative Studies on the production of Lipase by Bacillus species under various growth parameters. Int.J.Curr. Microbiol. App. Sci2 (11): 179-185.
[21]  Praveen, D. and Sharmishtha, P. (2011). Isolation, screening and production of extracullular alkaline lipase from a newly isolated Bacillus species. PD-12. Journal of Biological Sciences; 11(5): 381-387.
[22]  Kojima, Y., Kobayashi, M. and Shimizu, V. (2003). A Novel Lipase from Pseudomonas fluorescens HU380: Gene Cloning, Overproduction, Reneturation-Activation, Two-Step Purification, and Characterization. Journal of Bioscience and Bioengineering; 96(3): 242-249.
[23]  Dahiya, P., Arora, P., Chand, S., Chandhury, A. and Dilbaghi, N. (2010). Characterization of an extracellular alkaline lipase from Pseudomonas mondocinaM-37.Journal of Basic Microbiology. 50: 420-426.