来自大肠杆菌W菌株的生物降解型苏氨酸脱氨酶的克隆与过量表达。

Cloning and overproduction of biodegradative threonine deaminase from Escherichia coli W strain.

作者信息

Hirose K, Fujita M, Takeuchi M, Yumoto N, Tokushige M, Kawata Y

机构信息

Department of Chemistry, Faculty of Science, Kyoto University, Japan.

出版信息

Biotechnol Appl Biochem. 1992 Apr;15(2):134-41.

PMID:1586456

Abstract

We have cloned the structural gene (tdcB) of biodegradative threonine deaminase from Escherichia coli W strain by utilizing the polymerase chain reaction. The JM109/pUCTDA strain, which was obtained by transforming E. coli JM109 with a vector plasmid (pUCTDA) containing the cloned tdcB gene, produced a large amount of the enzyme corresponding to more than 5% of the total soluble protein. Amino acid sequence analysis of this recombinant enzyme showed that the amino acid sequence is identical to the nucleotide-deduced sequence of biodegradative threonine deaminase from E. coli K-12.

摘要

我们利用聚合酶链反应从大肠杆菌W菌株中克隆了生物降解性苏氨酸脱氨酶的结构基因（tdcB）。通过用含有克隆的tdcB基因的载体质粒（pUCTDA）转化大肠杆菌JM109获得的JM109/pUCTDA菌株产生了大量的该酶，其含量超过总可溶性蛋白的5%。对这种重组酶的氨基酸序列分析表明，其氨基酸序列与大肠杆菌K-12生物降解性苏氨酸脱氨酶的核苷酸推导序列相同。