Production of Lipase by Isolated Halophile, Halobacillus sp. Strain AR11 from International Miankaleh Wetland

Document Type : Research Article

Authors

1 Department of Microbiology, Faculty of Basic Science, University of Mazandaran, Babolsar, Iran

2 Department of Molecular and Cell Biology, Faculty of Basic Sciences, University of Mazandaran, Babolsar, Iran

3 Biochemistry Department, Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran

10.22080/jgr.2021.20826.1237

Abstract

Lipases are particularly important because they specifically hydrolyze acyl glycerol, oils, which are of great interest for different industrial applications. Some halophilic microorganisms produced some lipases. Halophilic bacteria have greater ability to produce salt and thermo tolerant enzymes like amylases, proteases and lipases. Most industrial enzymatic functions may be stopped by concentrated salt solutions and high temperatures; therefore, halophilic enzymes that have optimal activity at a vast range of temperature, pH and ionic strength, would be considered as suitable biocatalysts in industrial processes. The aim of this study was to isolate and study the halophilic lipase producing bacteria from the Miankaleh Wetland. The lipase activity was measured using titrimetric methods. Three halophilic strains (AR11, AR18 and AR28) were isolated from the Miankaleh wetland and were screened for the production of hydrolytic enzymes and lipolytic activity. Among three isolates, one strain was selected for identification using the molecular methods and some morphological characteristics. The bacterium Halobacillus truperi AR11 with 1.82 ± 0.1 U/mL lipase activity was selected as the highest lipase producing isolate. Lipase enzyme produced by this potential isolate was also characterized for determining its optimal activity. Effect of different pH, NaCl concentration and temperature on lipase activity was determined. The optimum pH for AR11 was found to be 9, while the optimum temperature and NaCl concentration for lipase activity was found to be 45ºC and 5 % NaCl, respectively. This is the first report of isolation and molecular identification of lipase producing halophilic bacteria from the Miankaleh wetland.

Keywords


Amoozegar MA, Malekzadeh F, Malik KA. 2003. Production of amylase by newly isolated moderate halophile Halobacillus sp. Strain MA-2. J Microbiol Method 52; 353-359.
Amoozegar MA, Schuman P, Hajighasemi M, Fatemi AZ. 2008.  Salinivibrio proteolyticus sp. nov., a moderately halophilic and proteolytic species from a hypersaline lake in Iran. Int J syst Evol Microbiol 58: 1159-1163.
Boutaiba S, Bhatnagar T, Hacene H, Mitchell DA, Baratti JC. 2006. Preliminary characterization of a lipolytic activity from an extremely halophilic archaeon, Natronococcus sp. J Mol Catal B Enzym 41: 21-26.
Castro-Ochoa L, Rodriguez-Gomez C, Valerio-Alfaro G, Oliart Ros R. 2005. Screening, purification and characterization of the thermo alkalophilic lipase produced by Bacillus thermoleovorans CCR11. Enzyme Microb Technol 37(6): 648-654.
Ghasemi Y, Rasoul-Amini S, Ebrahiminezhad A, Zarrini G, Kazemi A, Mousavi-Khorshidi S, Ghoshoon MB, Raee MJ. 2010. Halotolerant amylase production by a novel bacterial strain, Rheinheimera aquimaris. Res J Microbiol5(2): 144-149.
Govender L, Naidoo L, Setati ME. 2009. Isolation of hydrolase producing bacteria from Sua pan solar salterns and the production of endo-1, 4-β-xylanase from a newly isolated haloalkaliphilic Nesterenkonia sp. Afr J Biotechnol 8: 5458-5466.
Gutiérrez-Arnillas E, Rodríguez A, Sanromán MA, Deive FJ. 2016. New sources of halophilic lipases: isolation of bacteria from Spanish and Turkish saltworks. Biochem Eng J 109: 170-177.
Hasan F, Sanh A, Hameed A. 2005. Industrial applications of microbial lipases. Enzyme Microb Technol 39: 235-251.
José Murillo P, Barbosa Ranyere L, Souza Cláudia Moura de Melo Alini T, Mara F, Soares e Álvaro S. 2012.  Biochemical characterization of lipase from a new strain of Bacillus sp. ITP-001. Quim Nova 35(6): 1173-1178.
Kushner DJ. 1985. The Halobacteriacea, In the Bacteria, Vol.8, eds Woese .C.R. & Wolfe, R. S. Londen Academic Press. 171-214.
Kumar S, Kikon K, Upadhyay A, Kanwar S, Gupta R. 2005. Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulans BTS-3. Protein Expr Purif41(1): 38-44.
Mariana NG, Virginia A, Vargas HA, Michal S. 2008. Lipolytic enzyme production by halophilic/halotolerant microorganisms isolated from laguna verde, Bolivia. Rev Bol  Quim 25: 14-23.
Mobarak-Qamsari E, Kasra-Kermanshahi R, Moosavi-nejad Z. 2011. Isolation and identification of a novel, lipase-producing bacterium, Pseudomnas aeruginosa KM110. Iran J Microbiol 3(2): 92-98.
Niyonzima F, More S. 2014.  Biochemical properties of the alkaline lipase of Bacillus flexus XJU-1 and its detergent compatibility.  Biologia 69(9): 1108-1117.
Onishi H, Mori T, Takeuchi S, Tani K, Kobayashi T. 1983. Halophilic nuclease of moderately halophilic Bacillus sp .production purification and characteristic. Appl Environ Microbiol 45: 24-30.
Oren A. 2006. Life at High Salt Concentrations. Prokaryotes 2: 263-282.
Oren A. 2002. Molecular ecology of extremely halophilic archaea and bacteria. FEMS Microbiol Ecol 39: 1-7.
Ozcan B, Ozyilmaz G, Cokmus C, Caliskan M. 2009. Characterization of extracellular esterase and lipase activities from five halophilic archaeal strains. J Ind Microbiol. Biotech 36: 105-110.
Pignede G, Wang H, Fudalej F, Gaillardin C, Seman M, Nicaud  J. 2000. Characterization of an extracellular lipase encoded by LIP2 in Yarrowia lipolytica. J Bacteriol 182: 2802-2810.
Paul S, Bag SK, Das S, Harvill ET, Dutta C. 2008. Molecular signature of hyper saline adaptation: insights from genome and proteome composition of halophilic prokaryotes. Genome Biol 9: 1-19.
Rohban R, Amoozegar MA, Ventosa A. 2009. Screening and isolation of halophilic bacteria producing extracellular hydrolyses from Howz Soltan Lake, Iran. Ind Microbiol Biotechnol 36: 333-340.
Sahay H, Singh S, Kaushik R, Saxena AK, Arora DK. 2011. Characterization of halophilic bacteria from environmental samples from the brackish water of Pulicat Lake, India.  Biologia 66 (5): 741-747.
Schreck SD, Grunden AM. 2014. Biotechnological applications of halophilic lipases and thioesterases. Appl. Microbiol. Biotechnol 98: 1011-1021.
Soares MM, Silva RD, Gomes E. 1999. Screening of bacterial strains for pectinolytic activity: Characterization of the polygalacturonase produce by Bacillus sp. Rev Microbiol 30: 229-303.
Vidya J, Swaroop S, Singh SK, Alex D, Sukumaran RK, Pandey A. 2011. Isolation and characterization of a novelα-amylase from a metagenomic library of Western Ghats of Kerala, India. Biologia 66 (6): 939-944.
Weisburg WG, Barns SM, Pellriter DA, Lane DJ. 1991. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173: 697-703.
Kanlayakrit W, Boonpan A.  2007. Screening of halophilic lipase-producing bacteria and characterization of enzyme for fish sauce quality improvement. J Natural Sci41: 576- 585.