Cloning, expression, and identification of genes involved in the conversion of DL-2-amino-Delta2-thiazoline-4-carboxylic acid to L-cysteine via S-carbamyl-L-cysteine pathway in Pseudomonas sp. TS1138.
Two novel genes (tsB, tsC) involved in the conversion of DL-2-amino-Delta2-thiazoline-4-carboxylic acid (DL-ATC) to L-cysteine through S-carbamyl-L-cysteine (L-SCC) pathway were cloned from the genomic DNA library of Pseudomonas sp. TS1138. The recombinant proteins of these two genes were expressed in Escherichia coli BL21, and their enzymatic activity assays were performed in vitro. It was found that the tsB gene encoded an L-ATC hydrolase, which catalyzed the conversion of L-ATC to L-SCC, while the tsC gene encoded an L-SCC amidohydrolase, which showed the catalytic ability to convert L-SCC to L-cysteine. These results suggest that tsB and tsC play important roles in the L-SCC pathway and L-cysteine biosynthesis in Pseudomonas sp. TS1138, and that they have potential applications in the industrial production of L-cysteine.
Yu
Yangsheng
Y
Department of Microbiology, College of Life Sciences, Nankai University, Tianjin, P R China.
Liu
Zhong
Z
Liu
Chunqin
C
Li
Yang
Y
Jin
Yongjie
Y
Yang
Wenbo
W
Bai
Gang
G
D001483:Base Sequence; D002855:Chromatography, Thin Layer; D003001:Cloning, Molecular; D003545:Cysteine / Q000031:analogs & derivatives* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D015723:Gene Library; D005798:Genes, Bacterial; D007218:Industrial Microbiology / Q000379:methods; D008969:Molecular Sequence Data; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013844:Thiazoles / Q000378:metabolism*
D004269:DNA, Bacterial; D011994:Recombinant Proteins; D013844:Thiazoles; C015674:2-amino-delta(2)-thiazoline-4-carboxylic acid; C006894:S-carbamylcysteine; D003545:Cysteine