| Clostridium bolteae ATCC BAA-613 D-塔格糖3-差向异构酶的诱导表达、纯化及活性研究 |
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| 引用本文： | 储菲菲,沐万孟,邢庆超,周榴明,张涛,江波.Clostridium bolteae ATCC BAA-613 D-塔格糖3-差向异构酶的诱导表达、纯化及活性研究[J].食品工业科技,2012,33(7):198-201,392. |
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| 作者姓名： | 储菲菲 沐万孟 邢庆超 周榴明 张涛 江波 |
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| 作者单位： | 1. 江南大学食品科学与技术国家重点实验室,江苏无锡,214122
2. 罗盖特美国公司,基奥卡克52632,美国 |
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| 基金项目： | 国家自然基金项目(20906040);中央高校基本科研业务费专项资金(JUSRP31002);江苏省社会发展科技支撑项目(BE2010626) |
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| 摘 要： | D-塔格糖3-差向异构酶是生物法生产新型功能性因子D-阿洛酮糖最为有效的酶。一种新型的能够编码D-塔格糖3-差向异构酶的基因CLOBOL00069被克隆,它来源于Clostridium bolteae ATCC BAA-613。以pUC57为克隆载体,以pET-22b（+）为载体质粒,E.coli BL21（DE3）为宿主细胞,构建了基因重组工程菌。IPTG诱导剂诱导目的蛋白的表达;通过镍柱亲和层析,杂蛋白与目的蛋白得到了很好的分离。对纯化的重组蛋白样品进行SDS-PAGE分析,在约32ku处出现明显的特征条带。通过活性研究表明,Clostridium bolteae ATCC BAA-613 DTEase属于DTEase家族,并具有较高的生物转化率,反应10h后转化率达到20%。
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| 关 键 词： | D-塔格糖3-差向异构酶 D-阿洛酮糖 基因重组 亲和层析 |
Study on expression,purification and enzyme activity of Clostridium bolteae ATCC BAA-613 D-tagatose 3-epimerase |
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| CHU Fei-fei,MU Wan-meng,XING Qing-chao,ZHOU Liu-ming,ZHANG Tao,JIANG Bo.Study on expression,purification and enzyme activity of Clostridium bolteae ATCC BAA-613 D-tagatose 3-epimerase[J].Science and Technology of Food Industry,2012,33(7):198-201,392. |
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| Authors: | CHU Fei-fei MU Wan-meng XING Qing-chao ZHOU Liu-ming ZHANG Tao JIANG Bo |
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| Affiliation: | 1,(1.State Key Laboratory of Food Science and Technology,Jiangnan University,Wuxi 214122,China;2.Roquette America,Keokuk 52632,United States) |
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| Abstract: | D-tagatose 3-epimerase is the most effective enzyme for the biological production of D-psicose from D-fructose,a novel functional factor.Gene CLOBOL00069 encoding the D-tagatose 3-epimerase from Clostridium bolteae ATCC BAA-613 was cloned and expressed in Escherichia coli.E.coli BL21（DE3）were host cells with pUC57 as cloning vector and pET-22b（+）as plasmid to construct recombinant strains.The bacterium was induced by IPTG;then the recombinant DTEase was purified to electrophoretical homogeneity with affinity chromatography.The recombinant DTEase was analyzed by SDS-PAGE,and approximately 32ku exogenous protein was observed on the SDS-PAGE.The activity of recombinant DTEase was also studied,indicating that Clostridium bolteae ATCC BAA-613 DTEase belonged to DTEase family enzymes and had a high rate of biological transformation.The conversion reached 20% after 10h reaction. |
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| Keywords: | D-tagatose 3-epimerase D-psicose gene recombinant affinity chromatography |
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