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CLC number: Q26

On-line Access: 2011-07-04

Received: 2010-07-06

Revision Accepted: 2011-01-10

Crosschecked: 2011-06-16

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Journal of Zhejiang University SCIENCE B 2011 Vol.12 No.7 P.545-551


Construction of the yeast whole-cell Rhizopus oryzae lipase biocatalyst with high activity

Author(s):  Mei-ling Chen, Qin Guo, Rui-zhi Wang, Juan Xu, Chen-wei Zhou, Hui Ruan, Guo-qing He

Affiliation(s):  Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310029, China, School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China

Corresponding email(s):   ruanhui98@yahoo.com.cn

Key Words:  Rhizopus oryzae lipase (ROL), Yeast surface display, Codon optimization, Whole-cell biocatalyst

Mei-ling Chen, Qin Guo, Rui-zhi Wang, Juan Xu, Chen-wei Zhou, Hui Ruan, Guo-qing He. Construction of the yeast whole-cell Rhizopus oryzae lipase biocatalyst with high activity[J]. Journal of Zhejiang University Science B, 2011, 12(7): 545-551.

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author="Mei-ling Chen, Qin Guo, Rui-zhi Wang, Juan Xu, Chen-wei Zhou, Hui Ruan, Guo-qing He",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

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%T Construction of the yeast whole-cell Rhizopus oryzae lipase biocatalyst with high activity
%A Mei-ling Chen
%A Qin Guo
%A Rui-zhi Wang
%A Juan Xu
%A Chen-wei Zhou
%A Hui Ruan
%A Guo-qing He
%J Journal of Zhejiang University SCIENCE B
%V 12
%N 7
%P 545-551
%@ 1673-1581
%D 2011
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1000258

T1 - Construction of the yeast whole-cell Rhizopus oryzae lipase biocatalyst with high activity
A1 - Mei-ling Chen
A1 - Qin Guo
A1 - Rui-zhi Wang
A1 - Juan Xu
A1 - Chen-wei Zhou
A1 - Hui Ruan
A1 - Guo-qing He
J0 - Journal of Zhejiang University Science B
VL - 12
IS - 7
SP - 545
EP - 551
%@ 1673-1581
Y1 - 2011
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1000258

Surface display is effectively utilized to construct a whole-cell biocatalyst. codon optimization has been proven to be effective in maximizing production of heterologous proteins in yeast. Here, the cDNA sequence of Rhizopus oryzae lipase (ROL) was optimized and synthesized according to the codon bias of Saccharomyces cerevisiae, and based on the Saccharomyces cerevisiae cell surface display system with α-agglutinin as an anchor, recombinant yeast displaying fully codon-optimized ROL with high activity was successfully constructed. Compared with the wild-type ROL-displaying yeast, the activity of the codon-optimized ROL yeast whole-cell biocatalyst (25 U/g dried cells) was 12.8-fold higher in a hydrolysis reaction using p-nitrophenyl palmitate (pNPP) as the substrate. To our knowledge, this was the first attempt to combine the techniques of yeast surface display and codon optimization for whole-cell biocatalyst construction. Consequently, the yeast whole-cell ROL biocatalyst was constructed with high activity. The optimum pH and temperature for the yeast whole-cell ROL biocatalyst were pH 7.0 and 40 °C. Furthermore, this whole-cell biocatalyst was applied to the hydrolysis of tributyrin and the resulted conversion of butyric acid reached 96.91% after 144 h.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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