Isolation, Identification and Characterization of a Lipase Producing <i>Pseudomonas</i>

Saadatullah; Ijaz Malook; Mehmood Jan; Waheedullah; Noor Muhammmad; Zia ur Rehman

doi:doi:10.11648/j.jb.20180202.15

| Peer-Reviewed

Isolation, Identification and Characterization of a Lipase Producing Pseudomonas

Saadatullah, Ijaz Malook, Mehmood Jan, Waheedullah, Noor Muhammmad, Zia ur Rehman

Published in Journal of Biomaterials (Volume 2, Issue 2)

Received: 20 October 2018 Accepted: 9 November 2018 Published: 17 December 2018

Views: Downloads:

Download PDF

Share This Article

Twitter
Linked In
Facebook

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.

Published in	Journal of Biomaterials (Volume 2, Issue 2)
DOI	10.11648/j.jb.20180202.15
Page(s)	51-57
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2018. Published by Science Publishing Group

Keywords

Lipases, Pseudomonas, Production, Partial Purification, Molecular Characterization, Lipase Assay

References

[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.

Cite This Article

Plain Text BibTeX RIS

APA Style

Saadatullah, Ijaz Malook, Mehmood Jan, Waheedullah, Noor Muhammmad, et al. (2018). Isolation, Identification and Characterization of a Lipase Producing Pseudomonas. Journal of Biomaterials, 2(2), 51-57. https://doi.org/10.11648/j.jb.20180202.15

Copy | Download

ACS Style

Saadatullah; Ijaz Malook; Mehmood Jan; Waheedullah; Noor Muhammmad, et al. Isolation, Identification and Characterization of a Lipase Producing Pseudomonas. J. Biomater. 2018, 2(2), 51-57. doi: 10.11648/j.jb.20180202.15

Copy | Download

AMA Style

Saadatullah, Ijaz Malook, Mehmood Jan, Waheedullah, Noor Muhammmad, et al. Isolation, Identification and Characterization of a Lipase Producing Pseudomonas. J Biomater. 2018;2(2):51-57. doi: 10.11648/j.jb.20180202.15

Copy | Download

@article{10.11648/j.jb.20180202.15,
  author = {Saadatullah and Ijaz Malook and Mehmood Jan and Waheedullah and Noor Muhammmad and Zia ur Rehman},
  title = {Isolation, Identification and Characterization of a Lipase Producing Pseudomonas},
  journal = {Journal of Biomaterials},
  volume = {2},
  number = {2},
  pages = {51-57},
  doi = {10.11648/j.jb.20180202.15},
  url = {https://doi.org/10.11648/j.jb.20180202.15},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jb.20180202.15},
  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.},
 year = {2018}
}

Copy | Download

TY - JOUR
T1 - Isolation, Identification and Characterization of a Lipase Producing Pseudomonas
AU - Saadatullah
AU - Ijaz Malook
AU - Mehmood Jan
AU - Waheedullah
AU - Noor Muhammmad
AU - Zia ur Rehman
Y1 - 2018/12/17
PY - 2018
N1 - https://doi.org/10.11648/j.jb.20180202.15
DO - 10.11648/j.jb.20180202.15
T2 - Journal of Biomaterials
JF - Journal of Biomaterials
JO - Journal of Biomaterials
SP - 51
EP - 57
PB - Science Publishing Group
SN - 2640-2629
UR - https://doi.org/10.11648/j.jb.20180202.15
AB - 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.
VL - 2
IS - 2
ER -

Copy | Download

Author Information

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

Download PDF

Sections

Plain Text BibTeX RIS

APA Style

Saadatullah, Ijaz Malook, Mehmood Jan, Waheedullah, Noor Muhammmad, et al. (2018). Isolation, Identification and Characterization of a Lipase Producing Pseudomonas. Journal of Biomaterials, 2(2), 51-57. https://doi.org/10.11648/j.jb.20180202.15

Copy | Download

ACS Style

Saadatullah; Ijaz Malook; Mehmood Jan; Waheedullah; Noor Muhammmad, et al. Isolation, Identification and Characterization of a Lipase Producing Pseudomonas. J. Biomater. 2018, 2(2), 51-57. doi: 10.11648/j.jb.20180202.15

Copy | Download

AMA Style

Saadatullah, Ijaz Malook, Mehmood Jan, Waheedullah, Noor Muhammmad, et al. Isolation, Identification and Characterization of a Lipase Producing Pseudomonas. J Biomater. 2018;2(2):51-57. doi: 10.11648/j.jb.20180202.15

Copy | Download

@article{10.11648/j.jb.20180202.15,
  author = {Saadatullah and Ijaz Malook and Mehmood Jan and Waheedullah and Noor Muhammmad and Zia ur Rehman},
  title = {Isolation, Identification and Characterization of a Lipase Producing Pseudomonas},
  journal = {Journal of Biomaterials},
  volume = {2},
  number = {2},
  pages = {51-57},
  doi = {10.11648/j.jb.20180202.15},
  url = {https://doi.org/10.11648/j.jb.20180202.15},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jb.20180202.15},
  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.},
 year = {2018}
}

Copy | Download