| 大肠杆菌嗜盐α-淀粉酶基因的分子改造及其酶学特性 |
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| 引用本文： | 李剑龙,杨媛,吴昊,汤宏赤,韦宇拓.大肠杆菌嗜盐α-淀粉酶基因的分子改造及其酶学特性[J].广西科学,2016,23(1):25-30. |
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| 作者姓名： | 李剑龙 杨媛 吴昊 汤宏赤 韦宇拓 |
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| 作者单位： | 广西大学生命科学与技术学院,广西南宁530005;亚热带农业生物资源保护与利用国家重点实验室,广西南宁 530005 |
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| 基金项目： | 国家自然基金项目(31460437)，广西研究生教育创新计划项目(YCSZ2014050) |
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| 摘 要： | 【目的】对大肠杆菌Escherichia coli嗜盐α-淀粉酶基因进行改造,并探索嗜盐α-淀粉酶的嗜盐特性。【方法】从非嗜盐的大肠杆菌JM109中克隆到一个嗜盐α-淀粉酶基因k6并进行重组表达。通过同源建模,确定Na+结合位点上的氨基酸残基,并对相应位点进行定点突变。最后对突变酶的酶学性质进行研究。【结果】相对于野生酶,突变酶更加嗜盐,其最适NaCl浓度由2mol/L增加到3mol/L,最适pH值为7,最适温度为50℃,酶活力为4 831U/mg,提高近4倍。经HPLC检测,突变酶与2%(W/V)可溶性淀粉反应后的产物为葡萄糖、麦芽糖、麦芽三糖的混合物。【结论】嗜盐α-淀粉酶基因k6的嗜盐特性与Na+结合位点具有直接联系。
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| 关 键 词： | 非嗜盐菌 嗜盐淀粉酶 同源建模 定点突变 |
| 收稿时间： | 2015/11/20 0:00:00 |
| 修稿时间： | 2016/1/31 0:00:00 |
Escherichia coli Halophilic alpha Amylase Gene Molecular Modification and Enzymology Characteristics |
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| LI Jianlong,YANG Yuan,WU Hao,TANG Hongchi and WEI Yutuo.Escherichia coli Halophilic alpha Amylase Gene Molecular Modification and Enzymology Characteristics[J].Guangxi Sciences,2016,23(1):25-30. |
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| Authors: | LI Jianlong YANG Yuan WU Hao TANG Hongchi and WEI Yutuo |
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| Institution: | College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China,College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China and College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530005, China;State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Nanning, Guangxi, 530005, China |
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| Abstract: | |
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| Keywords: | non-halophilic bacteria halophilic amylase homology modeling site-directed mutagenesis |
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