[大肠杆菌tyrR基因敲除及其对苯丙氨酸生物合成的影响]。

Liang Shang, Chang-Sheng Fan, Rui-Liang Jin, Dong-Xin Liu, Jian-Gang Wang, Jun Yin, Da-Xin Song
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引用次数: 0

摘要

TyrR是由TyrR基因编码的全局调控蛋白,控制大肠杆菌中涉及芳香氨基酸生物合成和转运的几个转录单元。在这项工作中,tyrR基因通过双交叉同源重组被敲除。通过PCR和测序对该突变体进行了结构鉴定,并利用lacZ报告基因对其功能进行了验证。在tyrR-突变体中,受tyrR、DAHPS和AT控制的苯丙氨酸生物合成途径中两种关键酶的活性分别比亲本菌株提高了1.08倍和2.70倍。突变体苯丙氨酸生物合成产量是野生型菌株的1.59倍。对编码芳香氨基酸渗透酶的aroP的转录抑制被消除,导致tyrR-突变株芳香氨基酸运输增加70.2%。
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[Knockout of tyrR gene in Escherichia coli and its effects on the phenylalanine biosynthesis].

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.

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