文档介绍：
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#
郑春丽1,2,戴云杰1,马宏伟2,柳建设1**
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(1. 东华大学环境学院,上海 201620;
2. 内蒙古科技大学数理与生物工程学院,内蒙古包头 014010)
摘要: 丝氨酸乙酰转移酶( Serine acetyltransferase , SAT ) 是嗜酸氧化亚铁硫杆菌
(Acidithiobacillus ferrooxidans,A. ferrooxidans)菌中合成半胱氨酸的关键酶,SAT 在乙酰
-CoA 存在情况下催化 L-丝氨酸乙酰化生成 O-乙酰丝氨酸(OAS),随即 OAS 被催化生成
L-半胱氨酸。本文以 A. ferrooxidans 23270 标准菌株的基因组为模板, 通过 PCR 扩增
得到编码丝氨酸乙酰转移酶的基因,与原核表达载体 PLM 1 构建重组体,转入大肠杆菌
(Escherichia coli,E. coli)DH 5α中,测序正确后,加 IPTG 诱导表达, 用一步亲和层析法
纯化出了浓度和纯度都较高的丝氨酸乙酰转移酶。由紫外分析和 SDS-PAGE 确定其分子量
为 28 KDa,并成功测得其活性,比活为 ×104U/mg。
关键词:嗜酸氧化亚铁硫杆菌;丝氨酸乙酰转移酶;cysE;克隆;表达;纯化
中图分类号:Q89
Expression,Purification, and Characterization of a Serine
acetyltransferase from Acidithiobacillus Ferrooxidans
ZHENG Chunli1,2, DAI Yunjie1, MA Hongwei2, LIU Jianshe1
(1. College of Environmental Science and Engineering, Donghua University, ShangHai 201620;
2. School of Mathematics, Physics and Biological Engineering, Inner Mongolia University of
Science & Technology, BaoTou 014010)
Abstract: 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 23270 genomes was
template, the gene cysE which encodes Serine acetyltransferase was cloned, the fragment was
linked into the prokar