PURIFICATION AND CHARACTERIZATION OF EXTRACELLULAR LIPASE ENZYME FROM Aspergillus costaricaensis CBS115574
Asian Journal of Microbiology and Biotechnology, Volume 4, Issue 3,
Page 119-134
Abstract
In the present study, the lipase of alkaline nature obtained from the previously isolated strain Aspergillus costaricaensis CBS115574 was characterized and purified out using ammonium sulfate precipitation and chromatographic techniques on Diethylaminoethyl A-50 and Sephadex G-100. The purified lipase has the molecular weight of ~50 kDa which was determined by Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Zymography by using the phenol red along with the purification fold of 6.63 and specific activity of 31.73 IU/mg after the gel filtration chromatography. Lipase was found to be stabilized at the pH 7 (122.68 IU/g/min) and at the temperature of 80ºC (141.36 IU/g/min) with its optimal activity at pH 8 and temperature 50ºC. The Km and Vmax value for extracellular lipase were reported to be 29.62 mM and 74.07 IU/ml with their higher tolerance to glycerol (organic solvent), tween 80 (detergent), Ethylene diamine tetra acetic acid (activator & inhibitor), hydrogen peroxide (oxidizing & reducing agent) and sodium chloride (metal ions) with lipase activity of 106.74 IU/g/min, 84.53 IU/g/min, 108.06 IU/g/min, 97.17 IU/g/min and 118.7 IU/g/min. The above results suggested that A. costaricaensis CBS115574 lipase found a suitable position for application in different types of industries.
Keywords:
- Aspergillus costaricaensis CBS115574
- Diethylaminoethyl A-50
- Sephadex G-100; tolerance
How to Cite
CHAUDHARY, P., SHARMA, A. K., & ., P. (2020). PURIFICATION AND CHARACTERIZATION OF EXTRACELLULAR LIPASE ENZYME FROM Aspergillus costaricaensis CBS115574. Asian Journal of Microbiology and Biotechnology, 4(3), 119–134. Retrieved from https://ikprress.org/index.php/AJMAB/article/view/4853
References
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Andualema B and Gessesse A. Microbial lipase and their industrial applications: Review. Biotechnology. 2012;11(3):100-118.
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Penta JS, Johnson FM, Wachsman JT and Copeland WC. Mitochondrial DNA in human malignancy. Mutation Research. 2001;488(2): 119-33.
Avaiable:https://doi.org/10.1016/s1383-5742(01)00053-9
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Avaiable:https://doi.org/10.1155/2018/1618085
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Available:https://doi.org/10.7324/JAPS.2017.70108
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Sayali D, Sahithya K, Karthik L, Kumar G and Rao KVB. Isolation, purification and application of enzyme from Bacillus subtilis. Research Journal of Pharmaceuticals, Biological and Chemical Sciences. 2013; 4(3):1196-1204.
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Available: https://doi.org/10.5897/AJB10.369
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680-685.
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Available:https://doi.org/10.1080/13102818.2018.1446764
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Available: https://doi.org/10.1007/s11515-016-1409-z
Adetunji AI and Olaniran AO. Immobilization and characterization of lipase from an indigenous Bacillus aryabhattai SE3-PB isolated from the lipid-rich wastewater. Preparative Biochemistry and Biotechnology. 2018;48(10):898-905.
Available:https://doi.org/10.1080/10826068.2018.1514517
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Available:https://doi.org/10.1016/j.egypro.2014.07.293.
Chandorkar V, Gomashe AV and Parlewar S. Production of lipase from immobilized cells of Aspergillus niger. International Journal of Current Microbiology and Applied Sciences. 2014;3(8):703-707.
Sharma P, Sharma N, Sharma P, Pathania S and Handa S. Purification and characterization of a halotolerant and thermotolerant lipase produced from a novel bacterium “Brevibacterium halotolerans PS4 |KX671556|” and its application in detergent formulations. Proceedings of the Indian National Science Academy. 2017;83(3):681-687.
Available:https://doi.org/10.16943/ptinsa/2017/49025
Zouaoui B, Bouziane A and Ghalem BR. Isolation, purification and properties of lipase from Pseudomonas aeruginosa. African Journal of Biotechnology. 2012;11(60):12415-12421.
Available:https://doi.org/10.5897/AJB11.3951
Fatima H, Khan N, Rehman AU and Hussain Z. Production and partial characterization of lipase from Pseudomonas putida. Fermentation Technology. 2014;4(1):1-7.
Available:https://doi.org/10.4172/2167-7972-1000112
Das A, Shivakumar S, Bhattacharya S, Shakya S and Swathi SS. Purification and characterization of a surfactant-compatible lipase from Aspergillus tamarii JGIF06 exhibiting energy-efficient removal of oil stains from polycotton fabric. 3 Biotech. 2016;6:131.
Available: https://doi.org/10.1007/s13205-016-0449-z
Padhiar J, Das A and Bhattacharya S. Optimization of process parameters influencing the submerged fermentation of extracellular lipases from Pseudomonas aeruginosa, Candida albicans and Aspergillus flavus. Pakistan Journal of Biological Sciences. 2011; 14:1011-1018.
Mahmoud GA, Koutb MMM, Morsy FM and Bagy MMK. Characterization of lipase enzyme produced by hydrocarbons utilizing fungus Aspergillus terreus. European Journal of Biological Research. 2015; 5(3):70-77.
Barbosa JMP, Souza RL, de Melo, CM, Fricks AT, Soares CMF and Lima AS. Biochemical characterization of lipase from a new strain of Bacillus sp. ITP-001. Quimica nova. 2012;35(6):1-12.
Available:https://doi.org/10.1590/S0100-40422012000600020
Tiwari P and Upadhyay MK. Characterization of a thermo tolerant lipase from Pichia anamola. European Journal of Experimental Biology. 2012;2(3):464-467.
Jaiganesh R and Jaganathan MK. Isolation, purification and characterization of lipase from Bacillus sp. from kitchen grease. Asian Journal of Pharmaceutical and Clinical Research. 2018;11(6):224-227.
Ghori MI, Iqbal MJ and Hameed A. Characterization of a novel lipase from Bacillus sp. isolated from tannery wastes. 2011;42(1):1-10.
Available:https://doi.org/10.1590/S1517-83822011000100003
Kavitha M and Shanthi C. Alkaline thermostable cold active lipase from halotolerant Pseudomonas sp. VITCLP4 as detergent additive. Indian Journal of Biotechnology. 2017;16:446-455.
Ekinci AP, Dinçer B, Baltaş N, Adigüzel A. Partial purification and characterization of lipase from Geobacillus stearothermophilus AH22. Journal of Enzyme Inhibition and Medicinal Chemistry. 2015; 31(2):325-331. Avaiable:https://doi.org/10.3109/14756366.2015.1024677
Vorderwülbecke T, Epple M, Cammenga HK, Petersen S and Kieslich K. Racemic lactones from butterfat: an advanced approach that includes stereo differentiation. Journal of American Oil Chemists Society. 1992;69(8): 797-801.
Andualema B and Gessesse A. Microbial lipase and their industrial applications: Review. Biotechnology. 2012;11(3):100-118.
Available:https://doi.org/10.3923/biotech.2012.100.118
Penta JS, Johnson FM, Wachsman JT and Copeland WC. Mitochondrial DNA in human malignancy. Mutation Research. 2001;488(2): 119-33.
Avaiable:https://doi.org/10.1016/s1383-5742(01)00053-9
Kaja BS, Lumor S, Besong S, Taylor B and Ozbay G. Investigating enzyme activity of immobilized Candida rugosa lipase. Journal of Food Quality. 2018;1-9.
Avaiable:https://doi.org/10.1155/2018/1618085
Edupuganti S, Parcha L and Mangamoori LN. Purification and characterization of extracellular lipase from Staphylococcus epidermidis (MTCC 10656). Journal of Applied Pharmaceutical Science. 2017;7(1): 057-063.
Available:https://doi.org/10.7324/JAPS.2017.70108
Ananthi S, Ramasubburayan R, Palavesam A and Immanuel G. Optimization and purification of lipase through solid state fermentation by Bacillus cereus MSU as isolated from the gut of a marine fish Sardinella longiceps. International Journal of Pharmacy and Pharmaceutical Sciences. 2014;6(5):292-298.
Nath M, Hindumathy CK. Isolation, optimization and purification of lipase from Myroides odoratimimus. International Journal of Latest Research in Science and Technology. 2012;1(3):239-246.
Available:https://doi.org/10.29111
Dave BR, Sudhir AP and Subramanian RB. Purification and properties of an endoglucanase from Thermoascus aurantiacus. Biotechnology Reports. 2015;6:85-90.
Available:https://doi.org/10.1016/j.btre.2014.11.004
Lestari P, Raharjo TJ, Matsjeh S and Haryadi W. Partial purification and biochemical characterization of extracellular lipase from Azospirillum sp. JG3 bacteria. Advances of Science and Technology for Society. 2016; 080003(2016):080003-1- 080003-7.
Available: https://doi.org/10.1063/1.4958511.
Sayali D, Sahithya K, Karthik L, Kumar G and Rao KVB. Isolation, purification and application of enzyme from Bacillus subtilis. Research Journal of Pharmaceuticals, Biological and Chemical Sciences. 2013; 4(3):1196-1204.
Syed MN, Iqbal S, Bano S, Khan AB, Qader SAU and Azhar A. Purification and characterization of 60 KDa lipase linked with chaperonin from Pseudomonas aeruginosa BN-1. African Journal of Biotechnology. 2010; 9(45):7724-7732.
Available: https://doi.org/10.5897/AJB10.369
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680-685.
Borkar PS, Bodade RG, Rao SR and Khobragade CN. Purification and characterization of extracellular lipase from a new strain: Pseudomonas aeruginosa SRT9. Brazilian Journal of Microbiology. 2009;40(2): 358-366.
Available: https://doi.org/10.1590/S1517-838220090002000028
Hu J, Cai W, Wang C, Du X, Lin J and Cai J. Purification and characterization of alkaline lipase production by Pseudomonas aeruginosa HFE733 and application for biodegradation in food wastewater treatment. Biotechnology and Biotechnological Equipment. 2018;32(3):583-590.
Available:https://doi.org/10.1080/13102818.2018.1446764
Ugras S and Uzmez S. Characterization of a newly identified lipase from a lipase-producing bacterium. Frontiers in Biology. 2016;11(4): 323-330.
Available: https://doi.org/10.1007/s11515-016-1409-z
Adetunji AI and Olaniran AO. Immobilization and characterization of lipase from an indigenous Bacillus aryabhattai SE3-PB isolated from the lipid-rich wastewater. Preparative Biochemistry and Biotechnology. 2018;48(10):898-905.
Available:https://doi.org/10.1080/10826068.2018.1514517
Tripathi R, Singh J, Bharti RK and Thakur IS. Isolation, purification and characterization of lipase from Microbacterium sp. and its application in biodiesel production. Energy Procedia. 2014;54:518-529.
Available:https://doi.org/10.1016/j.egypro.2014.07.293.
Chandorkar V, Gomashe AV and Parlewar S. Production of lipase from immobilized cells of Aspergillus niger. International Journal of Current Microbiology and Applied Sciences. 2014;3(8):703-707.
Sharma P, Sharma N, Sharma P, Pathania S and Handa S. Purification and characterization of a halotolerant and thermotolerant lipase produced from a novel bacterium “Brevibacterium halotolerans PS4 |KX671556|” and its application in detergent formulations. Proceedings of the Indian National Science Academy. 2017;83(3):681-687.
Available:https://doi.org/10.16943/ptinsa/2017/49025
Zouaoui B, Bouziane A and Ghalem BR. Isolation, purification and properties of lipase from Pseudomonas aeruginosa. African Journal of Biotechnology. 2012;11(60):12415-12421.
Available:https://doi.org/10.5897/AJB11.3951
Fatima H, Khan N, Rehman AU and Hussain Z. Production and partial characterization of lipase from Pseudomonas putida. Fermentation Technology. 2014;4(1):1-7.
Available:https://doi.org/10.4172/2167-7972-1000112
Das A, Shivakumar S, Bhattacharya S, Shakya S and Swathi SS. Purification and characterization of a surfactant-compatible lipase from Aspergillus tamarii JGIF06 exhibiting energy-efficient removal of oil stains from polycotton fabric. 3 Biotech. 2016;6:131.
Available: https://doi.org/10.1007/s13205-016-0449-z
Padhiar J, Das A and Bhattacharya S. Optimization of process parameters influencing the submerged fermentation of extracellular lipases from Pseudomonas aeruginosa, Candida albicans and Aspergillus flavus. Pakistan Journal of Biological Sciences. 2011; 14:1011-1018.
Mahmoud GA, Koutb MMM, Morsy FM and Bagy MMK. Characterization of lipase enzyme produced by hydrocarbons utilizing fungus Aspergillus terreus. European Journal of Biological Research. 2015; 5(3):70-77.
Barbosa JMP, Souza RL, de Melo, CM, Fricks AT, Soares CMF and Lima AS. Biochemical characterization of lipase from a new strain of Bacillus sp. ITP-001. Quimica nova. 2012;35(6):1-12.
Available:https://doi.org/10.1590/S0100-40422012000600020
Tiwari P and Upadhyay MK. Characterization of a thermo tolerant lipase from Pichia anamola. European Journal of Experimental Biology. 2012;2(3):464-467.
Jaiganesh R and Jaganathan MK. Isolation, purification and characterization of lipase from Bacillus sp. from kitchen grease. Asian Journal of Pharmaceutical and Clinical Research. 2018;11(6):224-227.
Ghori MI, Iqbal MJ and Hameed A. Characterization of a novel lipase from Bacillus sp. isolated from tannery wastes. 2011;42(1):1-10.
Available:https://doi.org/10.1590/S1517-83822011000100003
Kavitha M and Shanthi C. Alkaline thermostable cold active lipase from halotolerant Pseudomonas sp. VITCLP4 as detergent additive. Indian Journal of Biotechnology. 2017;16:446-455.
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