Kosfeld Anne, Jahreis Knut
Centre for Pathology and Forensic and Genetic Medicine, Institute for Human Genetics-Hannover Medical School, Carl-Neuberg-Str.1, D-30625 Hannover, Germany.
Department of Biology and Chemistry, University of Osnabrück, Barbarastr.11, D-49069 Osnabrück, Germany.
Metabolites. 2012 Oct 16;2(4):756-74. doi: 10.3390/metabo2040756.
Escherichia coli is a widely used microorganism in biotechnological processes. An obvious goal for current scientific and technical research in this field is the search for new tools to optimize productivity. Usually glucose is the preferred carbon source in biotechnological applications. In E. coli, glucose is taken up by the phosphoenolpyruvate-dependent glucose phosphotransferase system (PTS). The regulation of the ptsG gene for the glucose transporter is very complex and involves several regulatory proteins. Recently, a novel posttranscriptional regulation system has been identified which consists of a small regulatory RNA SgrS and a small regulatory polypeptide called SgrT. During the accumulation of glucose-6-phosphate or fructose-6-phosphate, SgrS is involved in downregulation of ptsG mRNA stability, whereas SgrT inhibits glucose transport activity by a yet unknown mechanism. The function of SgrS has been studied intensively. In contrast, the knowledge about the function of SgrT is still limited. Therefore, in this paper, we focused our interest on the regulation of glucose transport activity by SgrT. We identified the SgrT target sequence within the glucose transporter and characterized the interaction in great detail. Finally, we suggest a novel experimental approach to regulate artificially carbohydrate uptake in E. coli to minimize metabolic overflow in biotechnological applications.
大肠杆菌是生物技术过程中广泛使用的微生物。该领域当前科学技术研究的一个明显目标是寻找优化生产力的新工具。通常,葡萄糖是生物技术应用中首选的碳源。在大肠杆菌中,葡萄糖通过磷酸烯醇丙酮酸依赖性葡萄糖磷酸转移酶系统(PTS)被摄取。葡萄糖转运蛋白的ptsG基因的调控非常复杂,涉及多种调控蛋白。最近,已鉴定出一种新型的转录后调控系统,其由小调控RNA SgrS和一种称为SgrT的小调控多肽组成。在6-磷酸葡萄糖或6-磷酸果糖积累期间,SgrS参与下调ptsG mRNA的稳定性,而SgrT通过一种尚不清楚的机制抑制葡萄糖转运活性。SgrS的功能已得到深入研究。相比之下,关于SgrT功能的了解仍然有限。因此,在本文中,我们将兴趣集中在SgrT对葡萄糖转运活性的调控上。我们确定了葡萄糖转运蛋白内的SgrT靶序列,并详细表征了其相互作用。最后,我们提出了一种新的实验方法来人工调控大肠杆菌中的碳水化合物摄取,以在生物技术应用中最小化代谢溢流。