High-level expression of Trigonopsis variabilis D-amino acid oxidase in Escherichia coli using lactose as inducer

被引：22

作者：

Hwang, TS

Fu, HM

Lin, LL

Hsu, WH ^{[1
]}

机构：

[1] Natl Chung Hsing Univ, Inst Mol Biol, Taichung 402, Taiwan

[2] Hung Kuang Inst Technol, Dept Food Sci & Nutr, Taichung 433, Taiwan

来源：

BIOTECHNOLOGY LETTERS | 2000年 / 22卷 / 08期

关键词：

D-amino acid oxidase gene; lactose induction; Trigonopsis variabilis;

D O I：

10.1023/A:1005647800700

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 use of lactose as inducer for the expression of Trigonopsis variabilis D-amino acid oxidase gene (daao) was investigated in Escherichia coli regulated by T7 or T5 promoter. The daao gene was prepared by reverse transcriptase-polymerase chain reaction and cloned into pET21b and pQE-30 to yield pET-DAAO and pQE-DAAO, respectively. The His(6)-tagged DAAO was expressed in E. coli and had a M-r value of approximately 39.3 kDa. In lactose-induced E. coli BL21 (DE3) (pET-DAAO), the expressed DAAO could comprise up to 15% of total soluble proteins and a productivity of 23.4 U ml(-1) was obtained.

引用

页码：655 / 658

页数：4

共 20 条

[1] Engineering the D-amino acid oxidase from Trigonopsis variabilis to facilitate its overproduction in Escherichia coli and its downstream processing by tailor-made metal chelate supports [J].