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Appl Microbiol Biotechnol. 2013 Aug;97(15):6677-83. doi: 10.1007/s00253-013-4988-5. Epub 2013 May 22.

Knocking out analysis of tryptophan permeases in Escherichia coli for improving L-tryptophan production.

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  • 1State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, People's Republic of China.

Abstract

Three permeases, Mtr, TnaB, and AroP, are involved in the uptake of L-tryptophan in Escherichia coli. These permeases possess individual function for cell transportation and metabolism, and affect extracellular L-tryptophan accumulation. In this study, by knocking out three tryptophan permeases separately and simultaneously in L-tryptophan-producing strain E. coli GPT1002, we analyzed the effect of permease knock out on L-tryptophan uptake, cell growth, and L-tryptophan production. We found that TnaB is the main transporter that is responsible for the uptake of L-tryptophan. Inactivation of tnaB improved the L-tryptophan production significantly, and inactivation of aroP has an additive effect on tnaB mutant. Quantitative real-time PCR analysis confirmed that knocking out permeases affects gene transcription and cell metabolism in many metabolic pathways. The tryptophan permease-deficient GPT1017 mutant exhibited the highest L-tryptophan production at 2.79 g l(-1), which is 51.6 % higher than that produced by the control strain. In 5-l bioreactor fermentation, the L-tryptophan production in GPT1017 reached 16.3 g l(-1).

PMID:
23695779
DOI:
10.1007/s00253-013-4988-5
[PubMed - indexed for MEDLINE]
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