嗜酸氧化亚铁硫杆菌丝氨酸乙酰转移酶的表达、纯化及性质鉴定

Serine acetyltransferase is a key enzyme involved in the pathway of the cysteine biosynthesis, catalyses the formation of O-acetyl-L-serine from acetyl-CoA and L-serine, then OAS be catalysed to L-cysteine. According to A. ferrooxidans ATCC 23270 genomes was template, the gene cysE which encodes Serine acetyltransferase was cloned, the fragment was linked into the prokaryotic expression vector PLM 1. Then the recombined expression plasmid was transduced into DH 5α. After sequence was mensurated right, with the addition IPTG was expressed in Escherichia coli, the soluble protein was purified by one-step affinity chromatography to apparent homogeneity. UV-vis scanning results of the recombinant protein. The molecular mass of the Serine acetyltransferase was 28 KDa by SDS-PAGE. The specific activity of the purified ATP sulfurylase was 8.0×104U/mg.