Purification and Characterization of Lipase produced by Pseudomonas aeruginosa isolated from some soils of Basrah Governorate
DOI:
https://doi.org/10.36320/ajb/v7.i2.8013Keywords:
Pseudomonas aeruginosa, lipase extractionAbstract
The present study aimed to Purification and characterization of lipase produced by the isolate Pseudomonas aeruginosa. The results showed that crude lipase was purified by using the precipitation step of several concentrations ratios of cold acetone. The concentration ratio 4:1 gave the best specific activity 6 U/mg of protein. The subsequent purification step using gel filtration was showed that specific activity reached to 40.5U/mg of protein. The gel electrophoresis (SDS-PAGE) of lipase purity were determined. the result showed there is an appearance of a single protein band predominant in all purification steps, While The molecular weight of purified lipase was 68KDaDownloads
References
Stoytcheva, M.; Montero, G.; Zlaler, R.; Leon, A. and Gochev, V. 2012. Analytical methods for lipase activity determination: A Review. Curr Ana. chem. 8, P: 400-407. DOI: https://doi.org/10.2174/157341112801264879
Lee, L.P.; Karbul, H. M.; Citartan, M.; Gopinath, S.C.; Lakshmipriya, T. and Tang, T. 2015. lipase secreting Bacillus species in an oil-contaminated Habitat promising strains to alleviate oil pollution. BioMed. Res Intern, P: 1-9. DOI: https://doi.org/10.1155/2015/820575
Bharathi, D.; Rajalakshmi, G.; Komathi, S.; Mythili, S.; Queen, J. and Sithara, N.V. 2016. Isolation, characterization and production of lipase producing bacterial strains from Petrol spilled soil. Intern J. Cur. Res.8, P: 36723-36726.
Sharma, S. and Kanwar, S.S. 2014. Organic solvent tolerant lipases and applications. The Scientific World. Available online: http://dx.doi.org/10.1155/2014/ 625258. P: 1-15 DOI: https://doi.org/10.1155/2014/625258
Borrelli, G.M. and Trono, D. 2015. Lipases and Phosphlipases and thier use as biocatalysts for industrial applications. Intern J. Mol Sci. 16, P: 20774-20840. DOI: https://doi.org/10.3390/ijms160920774
Padmapriya, B.; Rajeswari, T.; Noushida, E.; Sethupalan, D.G. Venil C.K. (2011). Production of lipase enzyme from Lactobacillus spp. and Its application in the degradation of meat. World Appl. Sci. J. 12, P: 1798-1802.
Jemli, S.; Ayadi-zouari, D.; Hlima, H.B. and Bejar, S. 2014. Biocatalysts , Application and engineering for industrial purposes. Crit Rev Biotechnol. 6, P: 1-13.
Yan, J.; Zheng, X.; Du, L. and Li, S. 2014. Integrated lipase production and in situ biodiesel synthesis in a recombinant Pichia pastotis yeast: an efficient dual bio catalytic system composed of cell catalysts. Biotechnol for biofu. 7(55), P: 1-8. DOI: https://doi.org/10.1186/1754-6834-7-55
Hasan, F.; Shah, A.A. and Hameed, A. 2009.Methods for detection and characterization of lipase: A comprehensive review, Biotecnol Adva. 27, P: 782-798. DOI: https://doi.org/10.1016/j.biotechadv.2009.06.001
Panesar, P.S.; Kaur, R.; Singla, G. and Sangwan, R.S. 2016. Bio-processing of Agro-industrial wastes for producing of food- grade enzymes: Process and prospects. Appli. food Biotechnol. 3(4), 208-227.
Svendsen, A. 2000. Lipase protein engineering: Review. Biochim et Biophysica Acta. 1543, P: 223-238. DOI: https://doi.org/10.1016/S0167-4838(00)00239-9
Gupta, R.; Gupta N.; Rathi P. 2004. Bacterial lipases: an overview of production, purification and biochemical properties. Appl Microbiol Biotechnol. 64, P: 763–781. DOI: https://doi.org/10.1007/s00253-004-1568-8
Aruna, K. and Khan, K. 2014. Optimization studies on production and activity of lipase obtained from Staphylococcus pasteuri SNA 59 isolated from spoit skin lotion. Intern. J. Cur. Microbio. and appl. sci. 3(5), P: 326-347.
Prasad, M.P. 2014. Production of lipase enzyme from Pseudomonas aeruginosa isolated from lipid rich soil. Intern J. Pure and Appl. Biosci. 2(1), P: 77-81.
Alhamdani, M.A. and Alkabbi, H.J.2017.Studyy on the activity and optimizationof lipase production bacteria isolated from oil contaminated soil. Indo- Asian J. of Multidisc. Res.3(1), P: 944- 956.
Zouaoui, B.; Bouziane, A. and Rahoghalem, B. 2012. Production optimization and purification of lipase from Pseudomonas aeruginosa. Afri. J. of Microbiol. Res. 6(20), P: 4417-4423. DOI: https://doi.org/10.5897/AJMR11.1567
Sagar, K.; Bashir,Y.; Phukan, M.M.; Konwar,B.K. 2013. Isolation Of Lipolytic Bacteria From Waste Contaminated Soil: A Study With Regard To Process Optimization For Lipase. Intern. J. Sci. and Technol. Res.2, P: 214 - 218.
Bradford, M.M.A. 1976. A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein dye binding . Anal Biochem. 72, P: 248-54. DOI: https://doi.org/10.1006/abio.1976.9999
Bhavani, M.; Chowdary, G.V.; David, M.; Archana, G. 2012. Screening, Isolation and Biochemical characterization of novel lipase producing from soil samples. Intern J. of Biol. Eng. 2(2), P: 18-22. DOI: https://doi.org/10.5923/j.ijbe.20120202.03
Ali, C.H.; Qureshi, A.; Mbadinga, S.M.; Lin, J.; Yang, S. and Mu, B. 2016. Organic solvent tolerant lipase from Pseudomonas aeruginosa FM-SH-1: Purification and characterization. JSM Enzymol. Protein Sci. 1(1), P: 1-7.
Al-Shammary, A.H.R. and Al-Husseiny, A.F.R. 2014. Extraction, Purification and Characterization of lipase produced by a local isolate of Staphylococcus aureus. Iraq J. Medi. Scie. 12(3), P: 253-260.
Amersham, 2002. Gel filtration: Principles and methods. Amersham Biosciences. P:1-120. DOI: https://doi.org/10.1016/S1365-6937(02)80019-3
Sambrook, J. and Russell, D.W. 2001. Molecular cloning. A Labratory Manual, 3rd edition. cold spring Harbor Laboratory press, cold spring Harbor NY, USA.
Bisht, D.; Yadav, S.K. and Darmwal, N.S. 2013. An oxidant and organic solvent tolerant alkaline lipase by P. aeruginosa mutant: downstream processing and biochemical characterization. Brazil J. Microbiol. 44(4), P: 1305–14. DOI: https://doi.org/10.1590/S1517-83822013000400040
Odunuga, O.O. and Shazhko, A. 2013. Ammonium sulfate precipitation combined with liquid chromatography is sufficient for purification of bovine serum albumin that is suitable for most routine laboratory applications. Biochemical compounds. http://www.hoajonline.com/Journals/pdf/2052-9341-13.pdf. P:1- 6. DOI: https://doi.org/10.7243/2052-9341-1-3
Adan, A. 2009. Isolation and identification of lipase producing psychrotrophic bacteria from soil: Cloning and partial characterization of its lipase, MSC thesis submitted to the Graduate School of Engineering and Sciences of Izmir Institute of Technology, Izmir university, Turkey. P: 1- 60.
Al-Ansari, B.M.M. 2016. Purification and Characterization of lipase extracted from soy bean seeds and used it as improved in baking. ph D thesis submitted to Agriculture collage , University of Basrah. P: 1- 210.
Costa-silva, T.A.; Souza, C.R.F.; Oliveira, W.P. and Siad, S. 2014. Charactrization and spray drying of lipase produced by the endophytic Fungus cercospora kikuchi. Brazi. J. Chem. Engin. 31(4), P: 849-858. DOI: https://doi.org/10.1590/0104-6632.20140314s00002880
Balcao, V.M.; Oliveira, T.A. and Malcata, F.X.C. 1997. Stability of a commercial lipase from Mucor Javanicum: Kinetic modeling of pH and temperature dependencies. Biocat. and Biotrans. 16, P: 45-66 DOI: https://doi.org/10.3109/10242429809040110
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2019 Hanaa jaffar jabbar Alkabee, Maytham Ayuob AlHamdani
This work is licensed under a Creative Commons Attribution 4.0 International License.
which allows users to copy, create extracts, abstracts, and new works from the Article, alter and revise the Article, and make commercial use of the Article (including reuse and/or resale of the Article by commercial entities), provided the user gives appropriate credit (with a link to the formal publication through the relevant DOI), provides a link to the license, indicates if changes were made and the licensor is not represented as endorsing the use made of the work.