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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/38448

Title: Recombinant Expression and Characterization of the Candida rugosa lip4 Lipase in Pichia pastoris: Comparison of Glycosylation, Activity and Stability
Authors: Shye-Jye Tang
Jei-Fu Shaw
Kuang-Hui Sun
Guang-Huan Sun
Terng-Yuan Chang
Ching-Kai Lin
Yuh-Chih Lo
Contributors: 國立臺灣海洋大學:生命科學暨生物科技學系
Keywords: Pichia pastoris
lipase
Candida rugosa
glycosylation
Date: 2001
Issue Date: 2016-09-06T01:32:34Z
Publisher: Archives of Biochemistry and Biophysis
Abstract: Abstract: Although Candida rugosa utilizes a nonuniversal serine codon (CUG) for leucine, it is possible to express lipase genes (LIP) in heterologous systems. After replacing the 19 CUG codons in LIP4 with serine codons by site-directed mutagenesis, a recombinant LIP4 was functionally overexpressed in Pichia pastoris in this study. This recombinant glycosylated lipase was secreted into the culture medium with a high purity of 100 mg/liter in a culture broth. Purified recombinant LIP4 had a molecular mass of 60 kDa, showing a range similar to that of lipase in a commercial preparation. Since LIP4 has only a glycosylation site at position Asn-351, this position may also be the major glycosylation site in C. rugosa lipases. Although the thermal stability of recombinant LIP4 significantly increased from 52 to 58°C after glycosylation, there were no significant differences in the catalytic properties of recombinant glycosylated lipase from P. pastoris and the unglycosylated one from Escherichia coil. These two recombinant LIP4s showed higher esterase activities toward long-chain ester (C16 and C18) and exhibited higher lipase activities toward unsaturated and long-chain lipids. In addition, LIP4 does not show interfacial activation as compared with LIP1 toward lipid substrates of tributyrin and triolein. These observations demonstrated that LIP4 shows distinguished catalytic activities with LIP1 in spite of their high sequence homology.
Relation: 187(1)
URI: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/38448
Appears in Collections:[生命科學系] 期刊論文

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