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Functional Expression of Candida antarctica Lipase A in Pichia a pastoris and Escherichia coli  

Park, Hye-Jung (Department of Chemical Engineering, Kwangwoon University)
Kim, Yong-Hwan (Department of Chemical Engineering, Kwangwoon University)
Publication Information
KSBB Journal / v.24, no.4, 2009 , pp. 341-346 More about this Journal
Abstract
Candida Antarctica lipase A (CalA) has been used because of its suitability in industrial applications. CalA has unique features capable to accept tertiary and sterically hindered alcohols among many hydrolases. CalA gene was cloned and constructed in expression vector such as pColdIII/CalA and $pPICZ{\alpha}A$/CalA. The gene encoding pColdIII/CalA was functionally expressed in the cytoplasm of Escherichia coli $Origami^{TM}$ B (DE3) cells. The plasmid $pPICZ{\alpha}A$/CalA linearized by BstX I was integrated into 5'AOX1 region of the chromosomal DNA and was functionally expressed in the methyl atrophic yeast Pichia pastoris. Expressed CalA in P. pastoris (0.7 Unit/mL) showed 35 times higher activity than that in E. coli expression system (0.02 Unit/mL).
Keywords
Candida antarctica lipase A; P. pastoris; E. coli; AOX 1;
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1 Svendsen, A. (2000), Lipase protein engineering, Biochim. Biophys. Act. 1543, 223-238   DOI   ScienceOn
2 Schmid, R. D. and R. Verger (1998), Lipases: interfacial enzymes with attractive applications, angew. Chem. Int. Ed. 37, 1608-1633   DOI   ScienceOn
3 Pfeffer, J., M. Rusnak, C. E. Hansen, R. B. Rhlid, R. D. Schmid, and S. C. Maurer (2007), Functional expression of lipase A from Candida antarctica in Escherichia coli : A prerequisite for high-throughput screening and directed evolution, J. Mol. Catal. 45, 62-67   DOI   ScienceOn
4 Krishna, S. H., M. Persson, and U. T. Bomschεuεr (2002), Enantioselective transεstεrification of a tertiary alcoh이 by lipase A from Candida antarctica, Tetrahed.: Asym. 13, 2693-2696   DOI   ScienceOn
5 Hohenblum, H., B. Gasser， M. Maurer, N. Borth, and D. Mattanovich (2004), Effects of gene dosage, promoters and substrates on unfolded protein stress of recombinant Pichia pastoris. Biotechnol. Bioeng. 85, 367-375   DOI   ScienceOn
6 Daly, R. and Milton T. W. Heam (2005), Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production, J. Mol. Recognit. 18, 119-138   DOI   ScienceOn
7 http://www.ebi.ac.uk/Tools/clustalw2
8 Ericsson, D. J., A. Kasrayan, P. Johanssoκ T. Bergfors, A. G. Sandstrom, J. E. Backvall, and S. L. Mowbray (2008), X-ray structure of Candida antarcticalipase A shows a novel lid structure and a likely mode of interfacial activation, J. Mol. Biol. 376(1), 109-19   DOI   ScienceOn
9 Xia, B., J. P. Etchegaray, and M. Inouye (2001), N onsense mutations in cspA cause ribosome trapping leading to complete growth inhibition and cell death at low temperature in Escherichia coli, J. Biol. Chem. 276, 35581-35588   DOI   ScienceOn
10 Pfeffer, J., S. Richter, J. Nieveler, C. E. Hansen, R. B. Rhlid, R. D. Schmid, and M. Rusnak (2006), High yield expression of Lipase A from Cαndida antarctica in the methylotrophic yeast Pichia pastorisand its purification and characterisation, Appl. Microbiol. Biotechnol. 72, 931-938   DOI   ScienceOn
11 Cristea, M., A. Engstrom, C. Su, L. Homsten, and E H. Oliw (2005), Expression of manganese lipoxygenase in Pichia pastoris and site-directed mutagenesis of putative metal ligands, Arch. Biochem. Biophys. 434, 201-211   DOI   ScienceOn
12 Cregg, J. M., S. V. Thomas, and C. R. William (1993), Recent advances in the expression of foreign genes in Pichia pastoris. Bio/Technol. 11, 905-910   DOI   ScienceOn
13 Hoegh, I., S. Patkar, T. Halkier, and M. T. Hansen (1995), Two lipases from Candida antarctica: cloning and expression in Aspergillus oryzae. Can. J. Bot. 73, 869-875   DOI   ScienceOn
14 Chen, J., Y. Q. Zhang, C. Q. Zhao, A. N. Li, Q. X. Zhou, and D. C. Li (2007), Cloning of a gene encoding thermostable glucoamylase from Chaetomium thermophilum and its expression in Pichia pastoris, J. Appl. Microbiol. 103, 2277-2284   DOI   ScienceOn
15 Macauley-Patrick, S., M. L. Fazenda, B. McNeil, and L. M. Harvey (2005), Heterologous protein production using the Pichia pastoris expression system, Yeast 22, 249-270   DOI   ScienceOn
16 Wang, H., S. Yin, X. Hou, and H. Xing (2005), Highlevel secretion expression of human ScFv against Botulinum neurotoxin A in Pichia pαstoris, Microbiol. in China 32, 50-53
17 Stephanie, J., E. Evguenieva-Hackenberg, and G. Klug (2004), Temperature-dependent processing of the cspA mRNA in Rhodobacter capsulatus, Microbiol. 150, 687-695   DOI   ScienceOn