Articles

Isolation and identification of a novel, lipase-producing bacterium, Pseudomnas aeruginosa KM110

Abstract

Background and Objectives: Lipases are particularly important due to the fact that they specifically hydrolyze acyl glycerol, oils and greases, which is of great interest for different industrial applications.
Materialst and Methods: In this study, several lipase-producing bacteria were isolated from wastewater of an oil processing plant. The strain possessing the highest lipase activity was identified both biochemically and sequencing of 16S rRNA gene. Then we increase lipase activity by improving conditions of production medium. Also, lipase from this strain was preliminarily characterized for use in industrial application.
Results: The 16S rRNA sequensing revealed it as a new strain of Pseudomonas aeruginosa and the type strain was KM110. An overall 3-fold enhanced lipase production (0.76 U mL-1) was achieved after improving conditions of production medium. The olive oil and peptone was found to be the most suitable substrate for maximum enzyme production. Also the enzyme exhibited maximum lipolytic activity at 45°C where it was also stably maintained. At pH 8.0, the lipase had the highest stability but no activity. It was active over a pH range of 7.0-10.0. The lipase activity was inhibited by Zn2+ & Cu2+ (32 and 27%, respectively) at 1mM. The enzyme lost 29% of its initial activity in 1.0% SDS concentration, whereas, Triton X-100, Tween-80 & DMSO did not significantly inhibit lipase activity.
Conclusions: Based on the findings of present study, lipase of P. aeruginosa KM110 is a potential alkaline lipase and a candidate for industrial applications such as detergent, leather and fine chemical industries.

Sharma R, Chisti Y, Banerjee UC. Production, purification, characterization, and applications of lipases. Biotechnol Adv 2001; 19: 627-662.

Saxena RK, Ghosh PK, Gupta R, Davinson WS, Bradoo S, Gulati R. Potential biocatalysis and future industry. Curr Sci 1999: 77: 110-115.

Sztajer H, Maliszewska I, Wieczorek J. Production of exogenous lipase by bacteria, fungi and actinomycetes.Enzyme Microb Technol 1998; 10: 492-7.

Gupta R, Gupta N, Rathi P. Bacterial lipases: an overview of production, purification and biotechnological properties. Appl Microbiol Biotechnol 2004; 64: 763-781.

Arpigny JL, Jaeger KE. Bacterial lipolytic enzymes:classification and properties. Biochem J 1999; 343:177-13.

Ertugrul S, Donmez G, Takac S. Isolation of lipase producing Bacillus sp. From olive mill waste water and improving its enzyme activity. J Hazard Mater 2007;149: 720-724.

Karadzic I, Masui A, Lidija Izrael Zivkovic LI, Fujiwara N. Purification and characterization of an alkaline lipase from Pseudomonas aeruginosa isolated from putrid mineral cutting oil as component of metal working fluid. J Biosci Bioeng 2006; 102: 82-89.

Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 2007; 24:1596-1599.

Thompson JD, Higgins DG, Gibson TJ. Clustal W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.Nucleic Acids Res 1994; 22: 4673-4680.

Lotti M, Monticelli S, Montesinos JL, Brocca S, Valero F, Lafuente J. Physiological control on the expression and secretion of Candida rugosa lipase. Chem Phys Lipids 1998; 93: 143-148.

Abdel-Fattah YR. Optimization of thermostable lipase production from a thermophilic Geobacillus sp. Using Box-Behnken experimental design. Biotechnol Lett 2002; 24: 1217-1222.

Kaushik R, Saran S, Isar J, Saxena RK. Statistical optimization of medium components and growth conditions by response surface methodology to enhance lipase production by Aspergillus carneus. J Mol Catal B-Enzym 2006; 40: 121-126.

He YQ, Tan TW. Use of response surface methodology to optimize culture medium for production of lipase with Candida sp. J Mol Catal 2006. B-Enzym., 43, 99-125.

Eltaweel MA, Rahman RNZRA, Salleh AB, Basri M.An organic solvent-stable lipase from Bacillus sp. Strain 42. Ann Microbiol 2005; 55: 187-192.

Lee DW, Koh YS, Kim KJ, Kim BC, Choi HJ, Kim DS, et al. Isolation and characterization of a thermophilic lipase from Bacillus thermoleovorans ID-1. FEMS Microbiol Lett 1999; 179: 393-400.

Chen J, Ishii T, Shimura S, Kirimura S, Usami S. Lipase production of Tricosporon fermentans WUC12 a newly isolated yeast. J Ferment Bioeng 1992; 73: 412-414.

Dharmsthiti S, Kuhasuntisuk B. Lipase from Pseudomonas aeruginosa LP602: Biochemical Properties Application for Wastewater Treatment. J Ind Microbiol Biotechnol 1998; 21: 75-80.

Lotrakul P, Dharmsthiti S. Lipase production by Aeromonas sobria LP004 in a medium containing whey and soybean meal. World J Microbiol Biotechnol 1997;13: 163-166.

Sharma R, Soni SK, Vohra RM, Jolly RS, Gupta LK,Gupta JK. Production of extracellular alkaline lipase from a Bacillus sp. RSJ1 and its application in ester hydrolysis. Ind J Microbiol 2002; 42: 49-54.

Sugihara A, Tani T, Tominaga Y. Purification and characterization of a novel thermostable lipase from Bacillus sp. J Biochem 1991; 109: 211-215.

Dong H, Gao S, Han SP, Cao SG. Purification and characterization of a Pseudomonas sp. lipase and its properties in non-aqueous media. Biotechnol App Biochem 1999; 30: 251-256.

Rathi P, Saxena RK, Gupta RA. Novel alkaline lipase from Bulkholderia cepacia for detergent formulation. Process Biochem 2001; 37: 187-192.

Kojima Y, Shimizu S. Purification and characterization of the lipase from Pseudomonas flurescens HU380. J Biosci Bioeng 2003; 96: 219-226.

Jinwal U, Roy U, Chowdhury A, Bhaduri A, Roy PK.Purification and characterization of an alkaline lipase from a newly isolated Pseudomonas mendoncina PK-12Cs and chemoselective hydolysis of fatty acid ester.Bioorgan Med Chem 2003; 11: 1041-1046.

Makhzoum A, Owusu-Apenten RK, Knapp JS.Purification and properties of lipase from Pseudomonas fluorescens strain 2D. Int Diary J 1996; 6: 459-472.

Svendsen A, Borch K, Barfoed M, Nielsen T, Gormsen E, Patkar S. Biochemical properties of cloned lipases from Pseudomonas family. Biochim Biophys Acta 1995;1259: 9-17.

Kanwar L, Goswami P. Isolation of Pseudomonas lipase produced in pure hydrocarbon substrate and its application in the synthesis of isoamyl acetate using membrane immobilized lipase. Enzyme Microb Technol 2002; 31: 727–735.

Shabtai Y, Daya-Mishne N. Production, purification and properties of lipase from a bacetrium (Ps. aeruginosa YS-7) capable of growing in water-restricted environments. Appl Environ Microbiol 1992; 58: 174-180.

Gilbert EJ, Cornish A, Jones C. Purification and proper- ties of extracellular lipase from Pseudomonas aerugino- sa EF2. J Gen Microbiol 1991b; 137: 2223-2229.

Chartrain M, Katz L, Marcin C, Thien M, Smith S, Fisher E, et al. Purification and characterization of a novel bioconverting lipase from Pseudomonas aeruginosa MB 5001. Enzyme Microb Technol 1993; 15: 575-580.

-Mencher JR, Alford JR. Purification and characteriza- tion of the lipase from Pseudomonas fragi. J Gen Microbiol 1967; 48: 317-328.

Matsumae H, Shibatani T. Purification and characterization of lipase from Serratia marcescens Sr41 8000 responsible for asymmetric hydrolysis of 3-phenylglycidic esters. J Ferment Bioeng 1994; 77:152-158.

Lelie D. Screening, purification and characteriza-tion of the thermoalkalophilic lipase produced by Bacillus thermoleovorans CCR11. Enzyme Microb Tech 2005;37: 648-654.

Schmidt-Dannert C, Sztajer H, Stocklein W, Menge U, Schmid RD. Screening, purification and properties of a thermophilic lipase from Bacillus thermocatenulatus. Biochim Biophys Acta 1994; 1214: 43-53.

Files
IssueVol 3 No 2 (2011) QRcode
SectionArticles
Keywords
Lipase Pseudomonas aeruginosa Stability

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Mobarak-Qamsari E, Kasra-Kermanshahi R, Moosavi-nejad Z. Isolation and identification of a novel, lipase-producing bacterium, Pseudomnas aeruginosa KM110. Iran J Microbiol. 1;3(2):92-98.