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谷氨酸棒杆菌中L-阿拉伯糖利用基因簇的鉴定与功能分析。

Identification and functional analysis of the gene cluster for L-arabinose utilization in Corynebacterium glutamicum.

作者信息

Kawaguchi Hideo, Sasaki Miho, Vertès Alain A, Inui Masayuki, Yukawa Hideaki

机构信息

Research Institute of Innovative Technology for the Earth, Kyoto, Japan.

出版信息

Appl Environ Microbiol. 2009 Jun;75(11):3419-29. doi: 10.1128/AEM.02912-08. Epub 2009 Apr 3.

Abstract

Corynebacterium glutamicum ATCC 31831 grew on l-arabinose as the sole carbon source at a specific growth rate that was twice that on d-glucose. The gene cluster responsible for l-arabinose utilization comprised a six-cistron transcriptional unit with a total length of 7.8 kb. Three l-arabinose-catabolizing genes, araA (encoding l-arabinose isomerase), araB (l-ribulokinase), and araD (l-ribulose-5-phosphate 4-epimerase), comprised the araBDA operon, upstream of which three other genes, araR (LacI-type transcriptional regulator), araE (l-arabinose transporter), and galM (putative aldose 1-epimerase), were present in the opposite direction. Inactivation of the araA, araB, or araD gene eliminated growth on l-arabinose, and each of the gene products was functionally homologous to its Escherichia coli counterpart. Moreover, compared to the wild-type strain, an araE disruptant exhibited a >80% decrease in the growth rate at a lower concentration of l-arabinose (3.6 g liter(-1)) but not at a higher concentration of l-arabinose (40 g liter(-1)). The expression of the araBDA operon and the araE gene was l-arabinose inducible and negatively regulated by the transcriptional regulator AraR. Disruption of araR eliminated the repression in the absence of l-arabinose. Expression of the regulon was not repressed by d-glucose, and simultaneous utilization of l-arabinose and d-glucose was observed in aerobically growing wild-type and araR deletion mutant cells. The regulatory mechanism of the l-arabinose regulon is, therefore, distinct from the carbon catabolite repression mechanism in other bacteria.

摘要

谷氨酸棒杆菌ATCC 31831能够以L-阿拉伯糖作为唯一碳源生长,其比生长速率是利用D-葡萄糖时的两倍。负责L-阿拉伯糖利用的基因簇包含一个六顺反子转录单元,全长7.8 kb。三个L-阿拉伯糖分解代谢基因,araA(编码L-阿拉伯糖异构酶)、araB(L-核酮糖激酶)和araD(L-核酮糖-5-磷酸4-表异构酶),组成了araBDA操纵子,在其上游以相反方向存在另外三个基因,araR(LacI型转录调节因子)、araE(L-阿拉伯糖转运蛋白)和galM(推定的醛糖1-表异构酶)。araA、araB或araD基因的失活消除了在L-阿拉伯糖上的生长,并且每个基因产物在功能上与其大肠杆菌对应物同源。此外,与野生型菌株相比,araE缺失突变体在较低浓度的L-阿拉伯糖(3.6 g L⁻¹)下生长速率降低>80%,但在较高浓度的L-阿拉伯糖(40 g L⁻¹)下没有降低。araBDA操纵子和araE基因的表达受L-阿拉伯糖诱导,并受到转录调节因子AraR的负调控。araR的破坏消除了在没有L-阿拉伯糖时的抑制作用。该调节子的表达不受D-葡萄糖的抑制,并且在需氧生长的野生型和araR缺失突变体细胞中观察到L-阿拉伯糖和D-葡萄糖的同时利用。因此,L-阿拉伯糖调节子的调节机制与其他细菌中的碳分解代谢物阻遏机制不同。

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