Cloning And Expression Of Conversion DL-2-amino-△～2-thiazoline-4-carboxylic Acid To L-cysteine In Pseudomonas Sp. TS1138

In our lab, a strain Pseudomonas sp. TS1138 was screened out which can convert DL-ATC to L-cysteine. Cloning and expression of genes of cysteine-forming enzymes and cysteine-degradating enzyme were studied in this paper. The results were as follows.From the genome random library of Pseudomonas sp.TS1138, the genes involved in conversion of DL-ATC to L-cysteine via the S-carbamoyl-L-cysteine (L-SCC) pathway were identified and cloned firstly. The ORFs were expressed in E. coli BL21 (DE3) by vector pET-21a (+), and the functions of recombinant protein were investigated by activity detection. The results indicated they were L-ATC hydrolase and L-SCC amidohydrolase genes. It was showed that the pathway of L-SCC as a intermediates in converting DL-ATC to L-cysteine existed in Pseudomonas strain.Furthermore, L-cysteine desulfhydrase gene, which was responsible for L-cysteine degradation, was cloned from TS1138. The capabilities of recombinant protein, expressed in E. coli BL21 (DE3), were visualized by activity staining.