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L-谷氨酰胺作为谷氨酸棒杆菌的氮源:AmtR 调控基因簇的去阻遏及其对氮感应的影响。

L-Glutamine as a nitrogen source for Corynebacterium glutamicum: derepression of the AmtR regulon and implications for nitrogen sensing.

机构信息

Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, D-91058 Erlangen, Germany.

Institut für Biotechnologie 1, Forschungszentrum Jülich, D-52425 Jülich, Germany.

出版信息

Microbiology (Reading). 2010 Oct;156(Pt 10):3180-3193. doi: 10.1099/mic.0.040667-0. Epub 2010 Jul 23.

Abstract

Corynebacterium glutamicum, a Gram-positive soil bacterium employed in the industrial production of various amino acids, is able to use a number of different nitrogen sources, such as ammonium, urea or creatinine. This study shows that l-glutamine serves as an excellent nitrogen source for C. glutamicum and allows similar growth rates in glucose minimal medium to those in ammonium. A transcriptome comparison revealed that the nitrogen starvation response was elicited when glutamine served as the sole nitrogen source, meaning that the target genes of the global nitrogen regulator AmtR were derepressed. Subsequent growth experiments with a variety of mutants defective in nitrogen metabolism showed that glutamate synthase is crucial for glutamine utilization, while a putative glutaminase is dispensable under the experimental conditions used. The gltBD operon encoding the glutamate synthase is a member of the AmtR regulon. The observation that the nitrogen starvation response was elicited at high intracellular l-glutamine levels has implications for nitrogen sensing. In contrast with other Gram-positive and Gram-negative bacteria such as Bacillus subtilis, Salmonella enterica serovar Typhimurium and Klebsiella pneumoniae, a drop in glutamine concentration obviously does not serve as a nitrogen starvation signal in C. glutamicum.

摘要

谷氨酸棒杆菌是一种革兰氏阳性土壤细菌,被广泛应用于各种氨基酸的工业生产中。它能够利用多种不同的氮源,如铵盐、尿素或肌酸。本研究表明,l-谷氨酰胺是谷氨酸棒杆菌的一种极好的氮源,其在葡萄糖最小培养基中的生长速度与铵盐相当。转录组比较显示,当谷氨酰胺作为唯一氮源时,会引发氮饥饿反应,这意味着全局氮调控因子 AmtR 的靶基因被去阻遏。随后在各种氮代谢缺陷突变体的生长实验中表明,谷氨酸合酶对于谷氨酰胺的利用至关重要,而在使用的实验条件下,一种假定的谷氨酰胺酶是可有可无的。编码谷氨酸合酶的 gltBD 操纵子是 AmtR 调控子的成员。在高细胞内 l-谷氨酰胺水平下引发氮饥饿反应的观察结果对氮感应具有重要意义。与枯草芽孢杆菌、鼠伤寒沙门氏菌和肺炎克雷伯菌等其他革兰氏阳性和革兰氏阴性细菌不同,在谷氨酸棒杆菌中,谷氨酰胺浓度的下降显然不能作为氮饥饿信号。

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