| Tyr78-loop对鱼腥藻来源苯丙氨酸脱氨酶活性的影响 |
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| 引用本文： | 郭军玲,张帆,黄楠,周丽,周哲敏.Tyr78-loop对鱼腥藻来源苯丙氨酸脱氨酶活性的影响[J].微生物学通报,2018,45(6):1171-1179. |
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| 作者姓名： | 郭军玲 张帆 黄楠 周丽 周哲敏 |
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| 作者单位： | 江南大学生物工程学院;江南大学工业生物技术教育部重点实验室 |
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| 基金项目： | 国家重点研发计划项目(2016YFE0127400)；中央高校基本科研业务费专项基金(JUSRP51713B) |
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| 摘 要： | 【背景】MIO(Methylidene-imidazol-5-one)依赖型酶中,催化因子Tyr所在Loop(Tyr78-loop)的灵活性显著影响酶学性质。【目的】探讨Tyr78-loop对鱼腥藻来源苯丙氨酸脱氨酶酶活的影响,以提高其反应活性。【方法】将该酶的Tyr78-loop进行分子改造,筛选出酶活提高的突变体,并对突变体的酶学性质进行研究。【结果】突变体S73N、E95V、E95K和S73N/E95K在37°C、pH 8.5下比活分别比原酶提高了34%、30%、18%和35%。蛋白三维结构模拟推测在突变体S73N、E95V和E95K中,位于α螺旋与Tyr78-loop交界处的Asn73、Val95和Lys95与附近氨基酸的氢键作用力数目减少,一定程度上增加了Tyr78-loop的柔性。【结论】Ser73位和Glu95位氨基酸的突变增加了Tyr78-loop的灵活性,提高了苯丙氨酸脱氨酶的酶活。
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| 关 键 词： | 苯丙氨酸脱氨酶，酶活，定点突变，稳定性 |
Influence of Tyr78-loop on phenylalanine ammonia-lyase from Anabaena variabilis |
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| GUO Jun-ling,ZHANG Fan,HUANG Nan,ZHOU Li and ZHOU Zhe-min.Influence of Tyr78-loop on phenylalanine ammonia-lyase from Anabaena variabilis[J].Microbiology,2018,45(6):1171-1179. |
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| Authors: | GUO Jun-ling ZHANG Fan HUANG Nan ZHOU Li and ZHOU Zhe-min |
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| Institution: | 1. School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China; 2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China,1. School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China; 2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China,1. School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China; 2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China,1. School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China; 2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China and 1. School of Biotechnology, Jiangnan University, Wuxi, Jiangsu 214122, China; 2. Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, Jiangsu 214122, China |
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| Abstract: | |
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| Keywords: | phenylalanine ammonia-lyase activity site-directed mutagenesis stability |
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