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Thermobifida fusca海藻糖合成酶的定点突变及其动力学性质研究
引用本文:王青艳,陈发忠,黄福宝,韦传东,韦宇拓,黄日波. Thermobifida fusca海藻糖合成酶的定点突变及其动力学性质研究[J]. 广西农业生物科学, 2007, 26(2): 115-119
作者姓名:王青艳  陈发忠  黄福宝  韦传东  韦宇拓  黄日波
作者单位:1. 广西大学,生命科学与技术学院,广西,南宁,530005
2. 南宁中诺生物工程有限责任公司,广西,南宁,530003
基金项目:国家973项目(2004CB719606)
摘    要:对Thermobifida fusca海藻糖合成酶(TreS)保守区域的氨基酸残基I224、N242、Q333、E352和N415进行定点突变,结果表明:位点I224、N242、N415突变后酶的活力与野生型TreS相近,E352位点突变后酶的比活力提高为野生型TreS的1.25倍。突变酶的最适温度、pH、Km和Kcat没有明显变化;突变Q333R则使TreS丧失了酶活力。

关 键 词:Thermobifida fusca  海藻糖合成酶  定点突变
文章编号:1008-3464(2007)02-0115-05
收稿时间:2006-04-27
修稿时间:2006-04-272006-11-09

Study on site-directed mutagenesis and kinetics of trehalose synthase from Thermobifida fusca
WANG Qing-yan,CHEN Fa-zhong,HUANG Fu-bao,WEI Chuan-dong,WEI Yu-tuo,HUANG Ri-bo. Study on site-directed mutagenesis and kinetics of trehalose synthase from Thermobifida fusca[J]. Journal of Guangxi Agricultural and Biological Science, 2007, 26(2): 115-119
Authors:WANG Qing-yan  CHEN Fa-zhong  HUANG Fu-bao  WEI Chuan-dong  WEI Yu-tuo  HUANG Ri-bo
Affiliation:1 College of Life Sciences and Technology, Guangxi University, Nanning 530005, China 2 Nanning Sinozyme Biotechnology Coporation Limited, Nanning 530003, China
Abstract:Five amino acid residues,I224,N242,Q333,E352 and N415,within the conserved region of TreS from Thermobifida fusca were subjected to site-directed mutagenesis,respectively .The results indicated that the TreS activities of the mutants derived from positions I224,N242 and N415 were similar to that of the wild type.The specific activity of the mutant E352R was 1.25-fold of the TreS of the wild type.The optimum pH and temperature for activity,Km and Kcat of TreS of these mutants did not significantly altered in comparison with those of TreS of the wild type .However,the mutant Q333R showed no TreS activity.
Keywords:Thermobifida fusca  trehalose synthase  site-directed mutagenesis
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