|本期目录/Table of Contents|

[1]史建明,徐兰兰,徐庆阳,等.谷氨酸棒杆菌YILW苏氨酸脱水酶基因的克隆表达及酶学性质[J].生物加工过程,2011,9(01):55-60.[doi:10.3969/j.issn.1672-3678.2011.01.012]
　SHI Jianming,XU Lanlan,XU Qingyang,et al.Expression and enzymatic properties of threonine dehydratase from Corynebacterium glutamicum YILW[J].Chinese Journal of Bioprocess Engineering,2011,9(01):55-60.[doi:10.3969/j.issn.1672-3678.2011.01.012]
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谷氨酸棒杆菌YILW苏氨酸脱水酶基因的克隆表达及酶学性质()
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《生物加工过程》[ISSN:1672-3678/CN:32-1706/Q]

卷:
9
期数:
2011年01期
页码:
55-60
栏目:
出版日期:
2011-01-20

文章信息/Info

Title:
Expression and enzymatic properties of threonine dehydratase from Corynebacterium glutamicum YILW
文章编号:
1672-3678(2011)01-0055-06
作者:
史建明; 徐兰兰; 徐庆阳; 谢希贤; 陈宁
天津科技大学生物工程学院工业微生物教育部重点实验室，天津300457
Author(s):
SHI Jianming; XU Lanlan; XU Qingyang; XIE Xixian; CHEN Ning
Key Laboratory of Industrial Microbiology of the Ministry of Education，College of Biotechnology，
Tianjing University of Science & Technology，Tianjing 300457，China
关键词:
谷氨酸棒杆菌; 酶学性质; 表达; 苏氨酸脱水酶
分类号:
Q812；Q78
DOI:
10.3969/j.issn.1672-3678.2011.01.012
文献标志码:
A
摘要:
将L异亮氨酸生产菌谷氨酸棒杆菌(Corynebacterium glutamicum YILW) 苏氨酸脱水酶（threonine dehydratase，TD）的编码基因ilvA在大肠杆菌中进行异源表达及进行初步的酶学性质研究。分别以C.glutamicum ATCC13032、YILW的基因组DNA为模板，利用PCR技术扩增出苏氨酸脱水酶的编码基因ilvA，测序获得编码序列。利用质粒PETHis将该基因在大肠杆菌BL21(DE3)中进行重组表达、金属螯合纯化，对其酶学性质进行初步研究。结果显示C. glutamicum YILW编码基因序列与已报道的ilvA序列相差5个碱基，相似度为996%，第383位氨基酸由苯丙氨酸突变为缬氨酸。酶学性质研究表明:重组酶YilwTD最适反应温度为32 ℃，在20~55 ℃范围内该酶较稳定，最适pH为67，该酶底物专一性强，对最适底物苏氨酸的米氏常数Km=832 mmol/L, 最大反应速度Vmax= 318×104  U/mg，与野生型酶相比,突变（F383V）后可显著降低终产物对酶的反馈抑制作用。为揭示突变对苏氨酸脱水酶活性的影响及进一步利用基因工程技术改造L异亮氨酸生产菌，提高L异亮氨酸产量奠定了基础。

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备注/Memo

备注/Memo:

收稿日期：2010-09-01
基金项目：国家高技术研究发展计划（863计划）资助项目(2007AA02Z228);天津市应用基础及前沿技术研究计划资助项目（08JCZDJC15400）
作者简介：史建明（1984—），男，河北沧州人，硕士研究生，研究方向：发酵代谢控制；陈宁（联系人），教授，博士生导师，Email:ningch@tust.edu.cn
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