Abstract
Two new lipases, LIP14 and LIP18, along with LIP8 from Yarrowia lipolytica MSR80 were functionally expressed as extracellular proteins with an IgG tag using Escherichia coli HB101 pEZZ18 host vector system. Each enzyme had an optimal activity at pH 7 and 40 °C and was activated by 6 mM Ca2+ and 90 % (v/v) non-polar solvents but inhibited by 10 mM of each 1,10-phenanthraline, DTNB, PMSF and N-bromosuccinamide. All the enzymes were thermostable with t1/2 of 52 min, 49 min and 68 min for LIP8, LIP14 and LIP18 at 80 °C, respectively. LIP18 was most thermostable among all with a high arginine: lysine ratio and proline content. All the three lipases showed a preference for oleic acid rich triacylglycerols and oils.
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Ficker P, Le DMT, Casaregola S, Gaillardin C, Thonart P, Nicaud JM (2005) Identification and charecterization of Lip7 and Lip8 genes encoding two extracellular triacylglycerol lipases in the yeast Yarrowia lipolytica. Fungal Genet Biol 42:264–274
Ficker P, Marty A, Nicaud TM (2011) The lipases from Yarrowia lipolytica: genetics, production, regulation, biochemical characterization and biotechnological applications. Biotechnol Adv 29:632–644
Guieysse D, Sandoval G, Faure L, Nicaud JM et al (2004) New efficient lipase from Yarrowia lipolytica for the resolution of 2-bromo-arylacetic acid esters. Tetrahedron Asym 14:317–323
Gupta N, Rathi P, Gupta R (2002) Simplified para-nitrophenyl palmitate assay for lipases and esterases. Anal Biochem 31:98–99
Harju S, Fedosyuk H, Peterson KR (2004) Rapid isolation of yeast genomic DNA: Bust n’ Grab. BMC Biotechnol 4:8
Kakugawa K, Shobayashii M, Suzuki O, Miyakawa T (2002) Purification and characterization of lipase from gycolipid-producing yeast Kurtzmanomyces sp. Biosci Biotechnol Biochem 66:978–985
Kumar SS, Arora N, Bhatnagar R, Gupta R (2009) Kinetic modulation of Trichosporon asahii MSR 54 lipase in presence of organic solvents: altered fatty acid specificity and reversal of enantio selectivity during hydrolytic reactions. J Mol Catal B Enzym 59:41–46
Liu WS, Pan XX, Jia B, Zhao HY, Xu L, Liu Y, Yan YJ (2010) Surface display of active lipases Lip7 and Lip8 from Yarrowia lipolytica on Saccharomyces cerevisiae. Appl Microbiol Biotechnol 88:885–891
Naka Y, Nakamura T (1992) The effect of serum albumin and related amino acid on pancreatic lipase and biles salts inhibited microbial lipases. Biosci Biotechnol Biochem 56:1066–1070
Olusesan AT, Azura LK, Forghani B, Bakar FA, Mohamed AKS, Radu S, Manap MYA, Saari N (2011) Purification, characterization and thermal inactivation kinetics of a non-regioselective thermostable lipase from a genotypically identified extremophilic Bacillus subtilis NS 8. New Biotechnol 6:738–745
Ortega N, Diego SD, Mateos MP, Busto MD (2004) Kinetic properties and thermal behaviour of polygalacturonase used in fruit juice clarification. Food Chem 88:209–217
Rudiger A, Jorgensen PL, Antranikan G (1995) Isolation and characterization of heat-stable pullulanase from the hyperthermophillic archaeon, Pyrococcus woesei after cloning and expression of its gene in E. coli. Appl Environ Microbiol 61:567–575
Sharma R, Gupta R (2010) Extracellular expression of Keratinase Ker P from Pseudomonas aeruginosa in E. coli. Biotechnol Lett 32:1863–1868
Syed MN, Iqbal S, Bano S, Khan AB, Qadan SAU, Azhar A (2010) Purification and characterization of 60 kDa lipase linked with chaperonin from Pseudomonas aeruginosa BN-1. Afr J Biotechnol 9:7724–7732
Szilagyi A, Zavodsky P (2000) Structural differences between mesophilic, moderately thermophilic and extremely thermophilic protein subunits: results of a comprehensive survey. Struct Fold Des 8:493–504
Yu MR, Qin SW, Tan TW (2007) Purification and characterization of the extracellular lipase Lip2 from Yarrowia lipolytica. Proc Biochem 42:384–391
Zhang Y, Menga K, Wang Y, Luo H, Yang P et al (2008) A novel proteolysis-resistant lipase from keratinolytic Streptomyces fradiae var. k11. Enzyme Microb Technol 42:346–352
Zhao H, Zheng L, Wang X, Liu Y, Xu L, Yan Y (2011) Cloning, expression and characterization of new lipases from Yarrowia lipolytica. Biotechnol Lett 33:2445–2452
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Authors acknowledge Council of Scientific and Industrial Research, India for financial assistance through project no. 38(1223)/09/EMR-II and R&D grant from Delhi University.
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Kumari, A., Gupta, R. Extracellular expression and characterization of thermostable lipases, LIP8, LIP14 and LIP18, from Yarrowia lipolytica . Biotechnol Lett 34, 1733–1739 (2012). https://doi.org/10.1007/s10529-012-0958-8
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DOI: https://doi.org/10.1007/s10529-012-0958-8