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通过表达聚羟基丁酸酯合成途径提高重组大肠杆菌中 L-色氨酸的产量。

The improved L-tryptophan production in recombinant Escherichia coli by expressing the polyhydroxybutyrate synthesis pathway.

机构信息

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

出版信息

Appl Microbiol Biotechnol. 2013 May;97(9):4121-7. doi: 10.1007/s00253-012-4665-0. Epub 2013 Jan 16.

Abstract

Polyhydroxybutyrate (PHB), the best known polyhydroxyalkanoates (PHA) has been believed to change intracellular metabolic flow and oxidation/reduction state, as well as enhance stress resistance of the host. In this study, a PHB biosynthesis pathway, which contains phaCAB operon genes from Ralstonia eutropha, was introduced into an L-tryptophan producing Escherichia coli strain GPT1002. The expression of the PHB biosynthesis genes resulted in PHB accumulation inside the cells and improved the L-tryptophan production. Quantitative real-time PCR analysis showed that the transcription of tryptophan operon genes in GPT2000 increased by 1.9 to 4.3 times compared with the control, indicating that PHB biosynthesis in engineered E. coli changed the physiological state of the host. Xylose was added into the medium as co-substrate to enhance the precursor supply for PHB biosynthesis. The addition of xylose improved both extracellular L-tryptophan production and intracellular PHB accumulation. Moreover, we obtained 14.4 g l(-1) L-tryptophan production and 9.7 % PHB (w/w) accumulation in GPT2000 via fed-batch cultivation.

摘要

聚羟基丁酸酯(PHB)是目前已知的聚羟基烷酸酯(PHA)中最好的一种,它被认为可以改变细胞内的代谢流和氧化还原状态,提高宿主的抗逆性。在本研究中,将来自恶臭假单胞菌的 phaCAB 操纵子基因引入到 L-色氨酸生产大肠杆菌菌株 GPT1002 中,构建了 PHB 生物合成途径。PHB 生物合成基因的表达导致细胞内 PHB 积累,并提高了 L-色氨酸的产量。定量实时 PCR 分析表明,与对照相比,工程大肠杆菌中 PHB 生物合成的色氨酸操纵子基因转录增加了 1.9 到 4.3 倍,表明 PHB 生物合成改变了宿主的生理状态。向培养基中添加木糖作为共底物,以增强 PHB 生物合成的前体供应。木糖的添加提高了细胞外 L-色氨酸的生产和细胞内 PHB 的积累。此外,通过分批补料培养,我们在 GPT2000 中获得了 14.4 g/L 的 L-色氨酸产量和 9.7%(w/w)的 PHB 积累。

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