Volume 2, Issue 2, December 2018, Page: 51-57
Isolation, Identification and Characterization of a Lipase Producing Pseudomonas
Saadatullah, Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Pakistan
Ijaz Malook, Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Pakistan
Mehmood Jan, Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Pakistan
Waheedullah, Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Pakistan
Noor Muhammmad, Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Pakistan
Zia ur Rehman, Department of Biotechnology and Genetic Engineering, Kohat University of Science and Technology, Kohat, Pakistan
Received: Oct. 20, 2018;       Accepted: Nov. 9, 2018;       Published: Dec. 17, 2018
DOI: 10.11648/j.jb.20180202.15      View  113      Downloads  25
Abstract
Lipases are hydrolytic enzymes and catalyze the hydrolysis of long-chain triacylglycerols into glycerol and fatty acid. Lipases are produced by plants, animals and microorganisms including bacteria and fungi. However microbial lipases, especially from bacteria, are more useful than their plant and animal derivatives because of several important properties. The primary goals of this research work is to isolate and identify a lipase producing Pseudomonas species from waste water samples collected from Dir lower, Peshawar and Kohat Khyber pakhtunkhwa Pakistan. The isolated bacteria were identified as Pseudomonas biochemically. The other purposes of this study are production, partial purification, characterization of lipase activity at different pH and incubation time, production and determination of molecular weight analysis. The lipase was partially purified up to 30% saturation using ammonium sulphate precipitation. Purity of lipases was cheeked by SDS-PAGE, showing the same banding pattern of all the lipases and the molecular weight were determined approximately 50kDA by comparing with protein marker bands. Spectrophotometric lipase assay was used for enzyme characterization. All the 7 isolates shows maximum activity at pH 7 after 48 hours of incubation and 37°C. Among all the isolates, isolate HSWPC shows highest activity of 110.11U /ml at pH 7 after 48 hours of incubation and 37°C. Pseudomonas lipases are widely used in food industry, detergent, paper, textile, leather and pharmaceutical industries because of their stability, selectivity and broad substrate specificity.
Keywords
Lipases, Pseudomonas, Production, Partial Purification, Molecular Characterization, Lipase Assay
To cite this article
Saadatullah, Ijaz Malook, Mehmood Jan, Waheedullah, Noor Muhammmad, Zia ur Rehman, Isolation, Identification and Characterization of a Lipase Producing Pseudomonas, Journal of Biomaterials. Vol. 2, No. 2, 2018, pp. 51-57. doi: 10.11648/j.jb.20180202.15
Copyright
Copyright © 2018 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Reference
[1]
Dong, H., S. Gao, S. Han and S. Cao (1990) Purification and characterization of a Pseudomonas sp. Lipase and its properties in non-aqueous media. Biotechnology Application Biochemistry. 30, 251.
[2]
Yamamoto, K. and N. Fujiwara (1995) The hydrolysis of castor oil using a lipase from Pseudomonas sp. f-B-24 positional and substrate specificity of the enzyme and optimum reaction conditions. Bioscience Biotechnology Biochemistry. 59, 1262.
[3]
Qamsari, E. M., R. K. Kermanshahi and Z. M. Nejad (2011) Isolation and identification of a novel, lipase producing bacterium, pseudomonas aeruginosa KM110. Iranian Journal of Microbiology. 3, 92.
[4]
Veerapagu, M., A. S. Narayanan, K. Ponmurugan and K. R. Jeya (2013) Screening selection identification production and optimization of bacterial lipases from oil spilled soil. Asian Journal of pharmaceutical and clinical research. 6, 62.
[5]
Rapp, P. and S. Backhaus (1992) Formation of extracellular lipases by filamentous fungi, yeast and bacteria. Journal of Enzyme Microbiology and Technology. 14, 938.
[6]
Vulfson, E. N., P. Woolley, S. B. Peterson (1994) Industrial applications of lipases their structure, biochemistry and applications. Cambridge University Press. 271.
[7]
Jaeger, K. E., B. W. Dijkstra and M. T. Reetz (1999) Bacterial biocatalysts molecular biology, Three-Dimensional Structures, and Biotechnological applications of Lipases. Annual Review of Microbiology. 53, 315.
[8]
Gokbulut, A. A. and Alper (2013) Purification and biochemical characterization of an extracellular lipase from psychrotolerant Pseudomonas fluorescens KE38. Turkish Journal of Biology. 37, 538.
[9]
Zouaoui, B., and A. Bouziane (2012) Production, optimization and characterization of the lipase from Pseudomonas aeruginosa. Romanian Biotechnological Letters. 17, 71-87.
[10]
Sharma, R., Y. Chisti and U. C. Banerjee (2001) Production, purification, characterization and applications of lipases. Biotechnology Advances. 19, 627.
[11]
Gupta, R., N. Gupta and P. Rathi (2004) Bacterial lipases an overview of production, purification and biochemical properties. Applied Microbial Biotechnology. 64, 763.
[12]
Stoyanova, V., G. Stanka , I. Ivan , M. Mariana and T. Sonya (2012) Lipolytic activity of genus Pseudomonas. JournalBioscience Biotechnology. 163-168.
[13]
Sirisha, E., M. L. Narasu and N. Rajasekar (2010) Isolation and Optimization of Lipase Producing Bacteria from Oil Contaminated Soils. Advances in Biological Research. 4, 249.
[14]
Underkofler, L. A., R. R. Barton, and S. S. Rennert (1957) Production of Microbial Enzymes and Their Applications. Application Microbiology. 3, 212.
[15]
Parsad, M. P., and K. Manjunath (2012) Effect of media and process parameters in the enhancement of extracellular lipase production by bacterial isolates from industrial effluents. International Journal of Microbiology Research. 4, 308.
[16]
Amara, A. A., and S. R. Salem (2009) Degradation of Castor Oil and Lipase Production by Pseudomonas aeruginosa. Journal of Agriculture and Environmental Science. 5,556.
[17]
Laemmli (1970) Cleavage of structural proteins during assembly of head of bacteriophage-T4. Nature. 227,680.
[18]
Krieger, N., A. S. Benjamin, C. R. Soccol, P. Nigam, and V. T. Soccol (1999) The realm of microbial lipases in biotechnology. Biotechnology Application Biochemistry. 29, 119.
[19]
Saeed, H. M., T. I. Zaghlolu, A. I. Khalil and M. T. Abdelbaeth (2005) purification and characterization of two extra cellular lipases from pseudomonas aeruginosa PS-X. Polish journal of microbiology. 54, 233.
[20]
Bornscheuer, U. T., C. Bessler, R. Srinivas and S. H. Krishna (2002) Optimizing lipases and related enzymes for efficient applications. Trends in Biotechnology. 20, 433.
[21]
Jaeger, K. E. and T. Eggert (2002) Lipases for biotechnology. Currurent Opinion in Biotechnology. 13, 390.
[22]
Chavez, G. S. and B. Palmeros (1994) Pseudomonas Lipases: Molecular Genetics and Potential Industrial Applications. Critical Review in Microbiology. 20, 95.
[23]
Joseph, B., P. W. Ramteke, G. Thomas and N. Shrivastava (2007) standard review cold-active microbial lipases, a versatile tool for industrial applications. Biotechnology and Molecular Biology Review. 2, 39.
[24]
Saxena, R. K., P. K. Ghosh, R. Gupta, W. S. Davidson, S. Bradoo, and R. Gulati (1999) Microbial lipases: Potential biocatalysts for the future industry. Current Science. 77, 101.
[25]
Xie, Y. C., H. Z. Liu and J. Y. Chen (1998) Candida rugosa lipase catalyzed esterification of racemic ibuprofen and chemical hydrolysis of S-ester formed. Biotechnology Letter. 20, 455.
[26]
Bergey, D. H. and R. S. Breed (1957) American Society for Microbiology. Bergey's manual of determinative bacteriology 7th edition.
[27]
Rashid, N., Y. Shimada, S. Ezaki, H. Atomi and T. Imanaka (2001) Low temperature lipase from psychrotrophic Pseudomonas species strain KB700A. Journal of Applied Environmental Microbiology. 67, 4064.
[28]
Stuer, W., K. E. Jaeger and U. K. Winkler (1986) Purification of extracellular lipase from Pseudomonas aeruginosa. Journal of Bacteriology. 168, 1070.
[29]
Tetling, S., U. K. Winkler, K. E. Jaeger and M. J. Benedik (1995) Gene cloning, sequence analysis, purification, and secretion by Escherichia coli of an extracellular lipase from Serratia marcescens. Application Environmental Microbiology. 61, 2674.
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