嗜酸氧化亚铁硫杆菌ATP硫酸化酶的表达、纯化及性质鉴定  

Expression,Purification and Characterization of ATP Sulfurylase from Acidithiobacillus ferrooxidans

作　　者：钱林[1] 郑春丽[1] 柳建设[1] 

机构地区：[1]东华大学环境科学与工程学院,上海201620

出　　处：《生物技术通报》2012年第6期136-140,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(50374075;41073060);上海市重点学科资助项目(B604);高等学校博士点专项科研基金(20100075110010)

摘　　要：ATP硫酸化酶是一种催化ATP和SO42-反应生成腺嘌呤-5'-磷酸硫酸(APS)和焦磷酸盐(PPi)的酶,它是硫酸根同化反应第一步的关键酶。以嗜酸氧化亚铁硫杆菌(A.ferrooxidansATCC 23270)基因组为模板,用PCR扩增得到ATPS基因,并克隆到表达载体pLM1上。加入IPTG的诱导表达,用AKTA蛋白纯化仪的镍柱亲和层析纯化得到浓度和纯度都较高的ATPS蛋白。SDS-PAGE分析,证实其分子量大小为33 kD,并成功的测出了其活性,比活达3.0×103U/mg。ATP sulfurylase, as a key enzyme that catalyzes the first step of the sulfur assimilation pathway, can catalyze the reaction between ATP and sulfate to produce APS and pyrophosphate （ PPi ） . In this study, the gene of ATP sulfurylase from A.ferrooxidans ATCC 23270 was cloned and expressed in Escherichia coli, and the protein was purified by AKTA explorer using affinity chromatography. SDS-PAGE revealed the molecular weight of ATP sulfurylase was 33 kD. The specific activity of the purified ATP sulfurylase was 3.0 x 103 U/mg.

关 键 词：嗜酸氧化亚铁硫杆菌 ATP硫酸化酶 表达 纯化 

分 类 号：X172[环境科学与工程—环境科学]