Browse > Article
http://dx.doi.org/10.48022/mbl.2109.09006

Purification, Characterization and Application of a Cold Active Lipase from Marine Bacillus cereus HSS  

Hassan, Sahar WM. (National Institute of Oceanography and Fisheries (NIOF))
Abd El Latif, Hala H. (National Institute of Oceanography and Fisheries (NIOF))
Beltagy, Ehab A. (National Institute of Oceanography and Fisheries (NIOF))
Publication Information
Microbiology and Biotechnology Letters / v.50, no.1, 2022 , pp. 71-80 More about this Journal
Abstract
Lipases (triacylglycerol acylhydrolases [EC 3.1.1.3]) are water-soluble enzymes. They catalyze the hydrolysis of fats and oils. A cold-active lipase from marine Bacillus cereus HSS, isolated from the Mediterranean Sea, Alexandria, Egypt, was purified and characterized. The total purification depending on lipase activity was 438.9 fold purification recording 632 U/mg protein. The molecular weight of the purified lipase was estimated to be 65 kDa using sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The optimum substrate concentration, enzyme concentration, pH, and temperature were 1.5 mM, 100 µl, pH 6 and 10℃, respectively. The lipase was tolerant to NaCl concentrations ranging from 1.5 to 4.5%. The lipase was affected by the tested metal ions, and its activity was inhibited by 16% in the presence of 0.05 M SDS. The application of the cold-active lipase for the removal of an oil stain from a white cotton cloth showed that it is a promising biological agent for the treatment of oily wastes and other related applications. To the best of our knowledge, this is the first report of the purification and characterization of a lipase from marine B. cereus HSS isolated from the Mediterranean Sea.
Keywords
Bacillus cereus HSS; lipase; purification; characterization; oil removal;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Heini H, Gebhardt R, Brecht A, Mecke D. 1987. Purification and characterization of rat liver glutaminase. Eur. J. Biochem. 162: 541-546.   DOI
2 Di Russo NV, Estrin DA, Marti MA, Roitberg AE. 2012. pH-dependent conformational changes in proteins and their effect on experimental pKas: the case of nitrophorin 4. PLoS Comput. Biol. 8: e1002761.   DOI
3 Afsar A, Cetinkaya F. 2008. A research on increasing the effectiveness of degreasing process by using enzymes. Textile Apparel 18: 278-283.
4 Javed S, Azeem F, Hussain S, Rasul I, Siddique MH, Riaz M, et al. 2017. Bacterial lipases: A review on purification and characterization. Progress Biophys. Mol. Biol. 132: 23-34.   DOI
5 Masomian M, Rahman RNZRA, Salleh AB, Basri M. 2010. A unique thermostable and organic solvent tolerant lipase from newly isolated Aneurinibacillusthermoaerophilus strain HZ: Physical factor studies. World J. Microbiol. Biotechnol. 26: 1693-1701.   DOI
6 Kumar S, Khyodano K, Ashutosh U, Shamsher S, Kanwar K, Gupta R. 2005. Production, purification, and characterization of lipase from thermophilic and alkaliphilic Bacillus coagulan BTS-3. Protein Expr. Purif. 41: 38-44.   DOI
7 Mazhar H, Abbas N, Ali SS, Hussain Z, Ali S. 2016. Purification and characterization of lipase production from Bacillus subtilis PCSIR-Nl39. J. Biol. Chem. Res. 33: 547-558.
8 Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the Folin-phenol reagent. J. Biol. Chem. 48: 265-275.
9 Najafi MF, Deobagkar D, Deobagkar D. 2005. Potential application of protease isolated from Pseudomonas aeruginosa PD100. Electron. J. Biotechnol., DOI: 10.2225/vol8-issue2-fulltext-5.   DOI
10 Kim YO, Khosasih V, Nam BH, Lee SJ, Suwanto A, Kim HK. 2011. Gene cloning and catalytic characterization of cold-adapted lipase of Photobacterium sp. MA1-3 isolated from blood clam. J. Biosci. Bioeng. 114: 589-595.   DOI
11 Laemmli UK. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.   DOI
12 Lineweaver H, Burk D. 1934. The determination of enzyme dissociation constants. J. Am. Chem. Soc. 56: 658-666.   DOI
13 Syifa F, Syihab SF, Madayanti F, Akhmaloka A, Widhiastuty MP. 2017. Purification and characterization of thermostable and alcohol tolerant lipase from Pseudoxanthomonas sp. Afr. J. Biotechnol. 16: 1670-1677.   DOI
14 Bae J, Kwon M, Kim I, Hou C, Kim H. 2014. Purification and characterization of a cold-active lipase from Pichialynferdii Y-7723: pH-dependent activity deviation. Biotechnol. Bioprocess Eng. 19: 851-857.   DOI
15 Filho DG, Silva AG, Guidini CZ. 2019. Lipases: sources, immobilization methods, and industrial applications. Appl. Microbiol. Biotechnol. 103: 7399-7423.   DOI
16 Jiewei T, Zuchao L, Peng Q, Lei W, Yongqiang T. 2014. Purification and Characterization of a cold-adapted lipase from Oceanobacillusstrain PT-11. PLoS One 9: e101343.   DOI
17 Gerday C, Aittaleb M, Arpigny JL, Baise E, Chessa E, Garssoux G, et al. 1997. Psychrophilic enzymes: a thermodynamic challenge. Biochim. Biophys. Acta 1342: 119-131.   DOI
18 Lanka S, Latha JN. 2015. Purification and characterization of a new cold active lipase, EnL A from Emericellanidulans NFCCI 3643. Afr. J. Biotechnol. 14: 1897-1909.
19 Sokalingam S, Madan B, Raghunathan G, Lee SG. 2013. Deciphering the factors responsible for the stability of a GFP variant resistant to alkaline pH using molecular dynamics simulations. Biotechnol. Bioprocess Eng. 18: 858-867.   DOI
20 Castro-Ochoa LD, Rodriguez-Gomez C, Valerio-Alfaro G, OliartRos R. 2005. Screening, purification and characterization of the thermoalkalophilic lipase produced by Bacillus thermoleovorans CCR11. Enzyme Microb. Technol. 37: 648-654.   DOI
21 Joseph B, Ramteke PW. 2013. Extracellular solvent stable cold-active lipase from psychrotrophic Bacillus sphaericus MTCC 7526: Partial purification and characterization. Ann. Microbiol. 63: 363-370.   DOI
22 Chaudhar P, Sharma AK, Precheta. 2020. Purification and characterization of extracellular lipase enzyme from Aspergillus costaricaensis CBS115574. Asian J. Microbiol. Biotechnol. 4: 119-134.
23 Sharma N, Pathania S, Handa S. 2017. Purification and characterization of lipase by Bacillus methylotrophicus PS3 under submerged fermentation and its application in detergent industry. J. Genet. Eng. Biotechnol. 15: 369-377.   DOI
24 Dharmsthiti S, Luchai S. 1999. Production, purification and characterization of thermophilic lipase from Bacillus sp. THL027. FEMS Microbiol. Lett. 179: 241-246.   DOI
25 Zhao J, Liu S, Gao Y, Ma M, Yan X, Cheng D, et al. 2021. Characterization of a novel lipase from Bacillus licheniformis NCU CS-5 for applications in detergent industry and biodegradation of 2,4-D butyl est. Int. J. Biol. Macromol. 176: 126-136.   DOI
26 Wang Q, Hou Y, Ding Y, Yan P. 2012. Purification and biochemical characterization of a cold-active lipase from Antarctic sea ice bacteria Pseudoalteromonas sp. NJ 70. Mol. Biol. Rep. 39: 9233-9238.   DOI
27 Kuddus M. 2018. Cold-active enzymes in food biotechnology: An updated mini review. J. Appl. Biol. Biotechnol. 6: 58-63.   DOI
28 Potumarthi R, Subhakar C, Vanajakshi J, Jetty A. 2008. Effect of aeration and agitation regimes on lipase production by newly isolated Rhodotorulamucilaginosa-MTCC 8737 in stirred tank reactor using molasses as sole production medium. Appl. Biochem. Biotechnol. 151: 700-710.   DOI
29 Awad G, Mostafa H, Danial EN, Abdelwahed NA, Awad HM. 2015. Enhanced production of thermostable lipase from Bacillus cereus ASSCRC-P1 in waste frying oil based medium using statistical experimental design. J. Appl. Pharm. Sci. 5: 7-15.
30 Joseph B, Ramteke PW, Kumar PA. 2006. Studies on the enhanced production of extracellular lipase by Staphylococcus epidermidis. J. Gen. Appl. Microbiol. 52: 315-320.   DOI
31 Bornscheuer UT. 1997. A reappraisal of the enzyme lipase for removing drying oil stains on paper. J. Inst. Pap. Conserv. 21: 37-47.   DOI
32 Jensen RG. 1983. Detection and determination of lipase (acylglycerol hydrolase) activity from various sources. Lipids 18: 650-657.   DOI
33 Muralidhar RV, Chirumamilla RR, Marchant R, Ramachandran VN, Ward OP, Nigam P. 2002. Understanding lipase stereo selectivity. World J. Microbiol. Biotechnol. 18: 81-97.   DOI
34 Hassan SWM, El Latif HHA, Ali SM. 2018. Production of cold-active lipase by free and immobilized marine Bacillus cereus HSS: Application in wastewater treatment. Front. Microbiol. 9: 2377.   DOI
35 Vakhlu J, Kour A. 2006. Yeast lipases: enzyme purification, biochemical properties and gene cloning. Elect. J. Biotechnol. 9: 69-85.   DOI
36 Treichel H, Oliveira DD, Mazuti MA, Luccio MD, Oliveira JV. 2010. A review on microbial lipases production. Food Bioproc. Technol. 3: 182-196.   DOI
37 Bornscheuer UT. 2018. Enzymes in lipid modification. Ann. Rev. Food Sci. Technol. 25: 85-103.   DOI
38 Salwoom L, Rahman RA, Zaliha RN, Salleh AB, Convey D, Pearce DM, et al. 2019. Isolation, characterisation, and lipase production of a cold-adapted bacterial strain Pseudomonas sp. LSK25 isolated from Signy Island, Antarctica. Molecules 24: 715-729.   DOI
39 Choo D, Kurihara T, Suzuki T, Soda KA. 1998. Cold-adapted lipase of an Alaskan gene cloning and enzyme purification and characterization. Appl. Environ. Microbiol. 64: 486-491.   DOI
40 Katiyar P, Pratibha, Hare V, Baghe VS. 2015. Isolation, partial purification and characterization of a cold active lipase from Pseudomonas sp., isolated from Satopanth Glacier of Western Himalaya, India. Int. J. Sci. Res. Manag. (IJSRM). 5: 6106-6112.
41 Obendorf SK, Varanasi A, Mejldal R, Thellersen M. 2001. Function of lipase in lipid soil removal as studied using fabrics with different chemical accessibility. J. Surfactants. Deterg 4: 233-245.   DOI
42 Ahmed HE, Kolisis FN. 2010. Enzymatic removal of the wasty dirt from a coptic tunic using the enzyme lipase. J. Text. Apparel. Technol. Manag. 6: 1-17.
43 Hertadi R, Widhyastuti H. 2015. Effects of calcium ion to the activity and stability of lipase isolated from Chromohalobacter japonicas BK-AB18. Procedia Chem. 16: 306-313.   DOI
44 Musa H, Hafiz Kasim F, Nagoor Gunny AA, Gopinath SC, Ahmad MA. 2018. Enhanced halophilic lipase secretion by Marinobacter-litoralis SW-45 and its potential fatty acid esters release. J. Basic Microbiol. 59: 87-100.   DOI