Beckers G, Nolden L, Burkovski A
Institut für Biochemie der Universität zu Köln, Zülpicher-Str. 47, D-50674 Köln, Germany.
Microbiology (Reading). 2001 Nov;147(Pt 11):2961-70. doi: 10.1099/00221287-147-11-2961.
The Corynebacterium glutamicum gltB and gltD genes, encoding the large (alpha) and small (beta) subunit of glutamate synthase (GOGAT), were investigated in this study. Using RT-PCR, a common transcript of gltB and gltD was shown. Reporter gene assays and Northern hybridization experiments revealed that transcription of this operon depends on nitrogen starvation. The expression of gltBD is under control of the global repressor protein AmtR as demonstrated by gel shift experiments and analysis of gltB transcription in an amtR deletion strain. In contrast to other bacteria, in C. glutamicum GOGAT plays no pivotal role; e.g. gltB and gltD inactivation did not result in growth defects when cells were grown in standard minimal medium and only a slight increase in the doubling time of the corresponding mutant strains was observed in the presence of limiting amounts of ammonia or urea. Additionally, mutant analyses revealed that GOGAT has no essential function in glutamate production by C. glutamicum.
本研究对谷氨酸棒杆菌中编码谷氨酸合酶(GOGAT)大亚基(α)和小亚基(β)的gltB和gltD基因进行了研究。通过逆转录聚合酶链反应(RT-PCR),显示出gltB和gltD的一个共同转录本。报告基因检测和Northern杂交实验表明,该操纵子的转录依赖于氮饥饿。凝胶迁移实验以及对amtR缺失菌株中gltB转录的分析表明,gltBD的表达受全局阻遏蛋白AmtR的控制。与其他细菌不同,在谷氨酸棒杆菌中GOGAT不发挥关键作用;例如,当细胞在标准基本培养基中生长时,gltB和gltD失活不会导致生长缺陷,并且在氨或尿素含量有限的情况下,仅观察到相应突变菌株的倍增时间略有增加。此外,突变分析表明,GOGAT在谷氨酸棒杆菌产生谷氨酸的过程中没有 essential 功能。(原文中“essential”未翻译,推测可能是笔误,根据语境这里可能是“重要”之类的意思,可根据实际情况调整)