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[大肠杆菌tyrR基因敲除及其对苯丙氨酸生物合成的影响]

[Knockout of tyrR gene in Escherichia coli and its effects on the phenylalanine biosynthesis].

作者信息

Shang Liang, Fan Chang-Sheng, Jin Rui-Liang, Liu Dong-Xin, Wang Jian-Gang, Yin Jun, Song Da-Xin

机构信息

Department of Microbiology, School of Life Sciences, Fudan University, Shanghai 200433,China.

出版信息

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai). 2003 Aug;35(8):728-33.

Abstract

TyrR is a global regulation protein encoded by tyrR gene which controls several transcriptional units involving biosynthesis and transportation of aromatic amino acids in Escherichia coli. In this work, the tyrR gene was knocked out by a double-cross homologous recombination. The tyrR- mutant was verified by structural identification by PCR and sequencing, and functional demonstration using lacZ reporter gene. In tyrR- mutant, the activities of two key enzymes in the phenylalanine biosynthesis pathway, whose expression was controlled by TyrR, DAHPS and AT, had been shown to elevate by a 1.08-fold and 2.70-fold compared with the parent strain, respectively. The yield of phenylalanine biosynthesis in the mutant was 1.59 times higher than that of wild type strain. The repression on the transcription of aroP, encoding an aromatic amino acid permease, was eliminated, resulting in a 70.2% increase of the aromatic amino acid transportation in tyrR- mutant strain.

摘要

TyrR是一种由tyrR基因编码的全局调控蛋白,它控制着大肠杆菌中几个涉及芳香族氨基酸生物合成和转运的转录单元。在这项工作中,通过双交叉同源重组敲除了tyrR基因。通过PCR和测序进行结构鉴定以及使用lacZ报告基因进行功能验证,证实了tyrR突变体。在tyrR突变体中,苯丙氨酸生物合成途径中两个关键酶(其表达受TyrR控制)DAHPS和AT的活性分别比亲本菌株提高了1.08倍和2.70倍。突变体中苯丙氨酸生物合成的产量比野生型菌株高1.59倍。对编码芳香族氨基酸通透酶的aroP转录的抑制作用被消除,导致tyrR突变体菌株中芳香族氨基酸转运增加了70.2%。

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