Cloning, expression, characterization and application of atcA, atcB and atcC from Pseudomonas sp. for the production of l-cysteine
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- 作者：Jingjing Duan (1) duankangjing@163.com
Qi Zhang (1) qizhang@nankai.edu.cn
Hongzhi Zhao (1) zhao_hong_zhi@126.com
Jun Du (23) dj@cifa.org.cn
Fang Bai (1) baifang1122@nankai.edu.cn
Gang Bai (12) gangbai@nankai.edu.cn - 关键词：dl ; 2 ; amino ; Δ ; 2 ; thiazoline ; 4 ; carboxylic acid – ; N ; carbamyl ; l ; cysteine pathway – ; l ; cysteine – ; Pseudomonas sp.
- 刊名：Biotechnology Letters
- 出版年：2012
- 出版时间：June 2012
- 年：2012
- 卷：34
- 期：6
- 页码：1101-1106
- 全文大小：251.8 KB
- 参考文献：1. Dhillon GS, Nagasawa T, Yamada H (1987) Microbial process for l-cysteine production. Enzyme Microb Tech 9:277–280
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8. Tashima I, Yoshida T, Asada Y, Ohmachi T (2006) Purification and characterization of a novell-2-amino-Δ2-thiazoline-4-carboxylic acid hydrolase from Pseudomonas sp. strain ON-4a expressed in E. coli. Appl Microbiol Biot 72:499–507
9. Yu YS, Liu Z, Liu CQ, Li Y, Jin YJ, Yang WB, Bai G (2006) Cloning, expression and identification of genes involved in the conversion of dl-2-amino-Δ2-thiazoline-4-carboxylic acid to l-cysteine via S-carbamyl-l-cysteine Pathway in Pseudomonas sp. TS1138. Biosci Biotech Bioch 70:2262–2267
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- 刊物类别：Biomedical and Life Sciences
- 刊物主题：Life Sciences
Microbiology
Biotechnology
Applied Microbiology
Biochemistry - 出版者：Springer Netherlands
- ISSN：1573-6776
文摘
An isolate of a Pseudomonas sp. uses the l-NCC (N-carbamoyl-l-cysteine) pathway to convert dl-2-amino-Δ2-thiazoline-4-carboxylic acid (dl-ATC) to l-cysteine. Genes encoding ATC racemase (AtcA), l-ATC hydrolase (AtcB) and l-NCC amidohydrolase (AtcC), involved in this pathway, were cloned from the Pseudomonas sp. and expressed in Escherichia coli BL21 via pET-28a(+). The resulting enzymes were purified, their functions identified, and their biochemical properties are described. In vitro catalysis experiments, using these enzymes, revealed that the bioconversion rate of l-cysteine from dl-ATC in the presence of AtcA was more efficient than in the absence of AtcA. This is the first report describing simultaneous cloning and expression of atcA, atcB and atcC and characterization of their enzymes for l-cysteine production from dl-ATC via the l-NCC pathway, enabling the complete l-NCC pathway to be elucidated.