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  • 莫莉,韦廷宗,闭海,郭媛,林丽华,黄日波,严少敏,庞浩.支链淀粉水解酶水解支链淀粉的特异氨基酸分析[J].广西科学,2015,22(1):31-36,43.    [点击复制]
  • MO Li,WEI Ting-zong,BI Hai,GUO Yuan,LIN Li-hua,HUANG Ri-bo,YAN Shao-min,PANG Hao.Analysis of the Key Amino Acid of Amylopectin Hydrolase on Amylopectin Hydrolysis[J].Guangxi Sciences,2015,22(1):31-36,43.   [点击复制]
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支链淀粉水解酶水解支链淀粉的特异氨基酸分析
莫莉1, 韦廷宗2, 闭海1, 郭媛2, 林丽华1,2, 黄日波1,2, 严少敏2, 庞浩1
0
(1.广西大学生命科学与技术学院, 广西南宁 530004;2.广西科学院, 非粮生物质酶解国家重点实验室, 国家非粮生物质能源工程技术研究中心, 广西生物质产业化工程院, 广西生物炼制重点实验室, 广西南宁 530007)
摘要:
[目的]通过对具有支链淀粉水解能力的环糊精水解酶的结构进行分析,探寻决定酶与支链淀粉水解相关的关键氨基酸残基。[方法]对底物特异性发生改变的环糊精水解酶cds1-3进行功能鉴定,并利用分子模拟方法对其底物作用特异氨基酸序列和空间结构进行分析。[结果]环糊精水解酶cds1-3具有特殊的底物作用方式,它水解支链淀粉的能力强于水解环糊精。cds1-3和ThMA的蛋白质序列只有30个氨基酸的差异,主要位于远离底物结合区域的蛋白质中段,与环糊精水解酶的底物通道聚集位置相近。[结论]环糊精水解酶cds1-3的功能鉴定及对其关键氨基酸进行序列和空间结构分析,为揭示大分子底物,特别是支链淀粉底物的水解方式提供新的切入点。
关键词:  环糊精水解酶  支链淀粉  酶水解  分子模拟
DOI:10.13656/j.cnki.gxkx.2015.01.001
投稿时间:2015-01-06修订日期:2015-01-16
基金项目:广西科技合作与交流计划项目(桂科合1347004-1),广西科技攻关项目(桂科重14122004-5),广西自然科学基金项目(2014GXNSFBA118129,2014GXNSFAA118078)和国家自然科学基金项目(21366007)资助。
Analysis of the Key Amino Acid of Amylopectin Hydrolase on Amylopectin Hydrolysis
MO Li1, WEI Ting-zong2, BI Hai1, GUO Yuan2, LIN Li-hua1,2, HUANG Ri-bo1,2, YAN Shao-min2, PANG Hao1
(1.College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530004, China;2.Guangxi Academy of Sciences, State Key Laboratory of Non-Food Biomass and Enzyme Technology, National Engineering Research Center for Non-food Biorefinery, Guangxi Biomass Industrialization Engineering Institute, Guangxi Key Laboratory of Biorefinery, Nanning, Guangxi, 530007, China)
Abstract:
[Objective] Exploring the key amino acid of amylopectin hydrolyzed enzyme, which have the ability of hydrolysis of huge substrate, is a possible way toward this problem.[Methods] A substrate-specificity mutant of cyclodextrin hydrolase was screened out from an environmental metagenomic library.Molecular simulation technique was applied to analyze the special amino acids related with substrate hydrolysis.[Results] Cyclodextrinhydrolase cds 1-3 showed a special mode of substrate hydrolytic action.The best substrate was amylopectin instead of cyclodextrin.This is different with other known cyclodextrin hydrolazed enzymes.[Conclusion] Cyclodextrinhydrolasecds 1-3 is a special cyclodextrin enzyme. The sequence and spatial structure analysis of its key amino acids may provide a good basis for hydrolysis of amylopectin.
Key words:  cyclodextrin hydrolase  amylopectin  enzymatic hydrolysis  molecular simulation

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