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对一种代谢酶和一种转录调节因子进行基因操作以增加单细胞蓝细菌的琥珀酸排泄量。

Genetic manipulation of a metabolic enzyme and a transcriptional regulator increasing succinate excretion from unicellular cyanobacterium.

作者信息

Osanai Takashi, Shirai Tomokazu, Iijima Hiroko, Nakaya Yuka, Okamoto Mami, Kondo Akihiko, Hirai Masami Y

机构信息

RIKEN Center for Sustainable Resource Science Yokohama, Japan ; Department of Agricultural Chemistry, School of Agriculture, Meiji University Kawasaki, Japan.

Biomass Engineering Program, RIKEN Yokohama, Japan.

出版信息

Front Microbiol. 2015 Oct 6;6:1064. doi: 10.3389/fmicb.2015.01064. eCollection 2015.

Abstract

Succinate is a building block compound that the U.S. Department of Energy (DOE) has declared as important in biorefineries, and it is widely used as a commodity chemical. Here, we identified the two genes increasing succinate production of the unicellular cyanobacterium Synechocystis sp. PCC 6803. Succinate was excreted under dark, anaerobic conditions, and its production level increased by knocking out ackA, which encodes an acetate kinase, and by overexpressing sigE, which encodes an RNA polymerase sigma factor. Glycogen catabolism and organic acid biosynthesis were enhanced in the mutant lacking ackA and overexpressing sigE, leading to an increase in succinate production reaching five times of the wild-type levels. Our genetic and metabolomic analyses thus demonstrated the effect of genetic manipulation of a metabolic enzyme and a transcriptional regulator on succinate excretion from this cyanobacterium with the data based on metabolomic technique.

摘要

琥珀酸是一种结构单元化合物,美国能源部(DOE)已宣布其在生物炼制中很重要,并且它被广泛用作一种商品化学品。在此,我们鉴定出了两个可提高单细胞蓝藻聚球藻属PCC 6803琥珀酸产量的基因。琥珀酸在黑暗、厌氧条件下分泌,通过敲除编码乙酸激酶的ackA以及过表达编码RNA聚合酶σ因子的sigE,其产量水平得以提高。在缺乏ackA且过表达sigE的突变体中,糖原分解代谢和有机酸生物合成增强,导致琥珀酸产量增加,达到野生型水平的五倍。因此,我们的遗传和代谢组学分析利用代谢组学技术的数据,证明了对一种代谢酶和一种转录调节因子进行基因操作对这种蓝藻琥珀酸分泌的影响。

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