| 嗜酸氧化亚铁硫杆菌ATP硫酸化酶的表达、纯化及性质鉴定 |
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| 引用本文： | 钱林,郑春丽,柳建设.嗜酸氧化亚铁硫杆菌ATP硫酸化酶的表达、纯化及性质鉴定[J].生物技术通报,2012(6):136-140. |
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| 作者姓名： | 钱林 郑春丽 柳建设 |
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| 作者单位： | 东华大学环境科学与工程学院,上海,201620 |
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| 基金项目： | 国家自然科学基金，上海市重点学科资助项目，高等学校博士点专项科研基金 |
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| 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。
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| 关 键 词： | 嗜酸氧化亚铁硫杆菌 ATP硫酸化酶 表达 纯化 |
Expression, Purification and Characterization of ATP Sulfurylase from Acidithiobacillus ferrooxidans |
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| Qian Lin
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Zheng Chunli
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Liu Jianshe.Expression, Purification and Characterization of ATP Sulfurylase from Acidithiobacillus ferrooxidans[J].Biotechnology Bulletin,2012(6):136-140. |
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| Authors: | Qian Lin Zheng Chunli Liu Jianshe |
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| Institution: | Qian Lin Zheng Chunli Liu Jianshe(College of Environmental Science and Engineering,Donghua University,Shanghai 201620) |
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| Abstract: | 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 3.0×103?U/mg. |
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| Keywords: | Acidithiobacillus ferrooxidans ATP sulfurylase Expression Purification |
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