High-level expression of the Penicillium notatum glucose oxidase gene in Pichia pastoris using codon optimization

被引：54
|
作者：
Gao, Zhaowei [1 ,2 ]
Li, Zhuofu [3 ]
Zhang, Yuhong [1 ]
Huang, Huoqing [4 ]
Li, Mu [1 ]
Zhou, Liwei [1 ]
Tang, Yunming [2 ]
Yao, Bin [4 ]
Zhang, Wei [1 ]
机构：
[1] Chinese Acad Agr Sci, Biotechnol Res Inst, Beijing 100081, Peoples R China
[2] Southwest Univ, Sch Life Sci, Chongqing 400715, Peoples R China
[3] Changchun Univ Sci & Technol, Changchun 130022, Jilin, Peoples R China
[4] Chinese Acad Agr Sci, Key Lab Feed Biotechnol, Minist Agr, Feed Res Inst, Beijing 100081, Peoples R China
来源：
BIOTECHNOLOGY LETTERS | 2012年 / 34卷 / 03期
关键词：
Codon optimization; Glucose oxidase; Penicillium notatum; Pichia pastoris; ASPERGILLUS-NIGER; BREAD; PCR;
D O I：
10.1007/s10529-011-0790-6
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
The glucose oxidase (GOD) gene from Penicillium notatum was expressed in Pichia pastoris. The 1,815 bp gene, god-w, encodes 604 amino acids. Recombinant GOD-w had optimal activity at 35-40 degrees C and pH 6.2 and was stable, from pH 3 to 7 maintaining >75% maximum activity after incubation at 50 degrees C for 1 h. GOD-w worked as well as commercial GODs to improve bread making. To achieve high-level expression of recombinant GOD in P. pastoris, 272 nucleotides involving 228 residues were mutated, consistent with the codon bias of P. pastoris. The optimized recombinant GOD-m yielded 615 U ml(-1) (2.5 g protein l(-1)) in a 3 l fermentor-410% higher than GOD-w (148 U ml(-1)), and thus is a low-cost alternative for the bread baking industry.
引用
收藏
页码：507 / 514
页数：8
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