Abstract

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.

Highlights

  • With its fast growth and simple cultivation Escherichia coli is a widely used microorganism in biotechnological processes and in industrial microbiology

  • In order to test the assumption of a direct protein–protein interaction between SgrT and EIICBGlc, we performed an in vivo crosslinking experiment with paraformaldehyde

  • To identify the two interaction partners in subsequent. Both proteins were tagged with different flags (EIICBGlc-5His, SgrT-3HA)

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Summary

Introduction

With its fast growth and simple cultivation Escherichia coli is a widely used microorganism in biotechnological processes and in industrial microbiology. Different growth strategies [3] have been applied as well as targeted changes in central carbon metabolism [2,4] or control of the glucose transport process has been modified [5,6]. The latter approach seems to be very helpful since acetate excretion mainly occurs when the transport rate exceeds the metabolism which causes a temporal metabolic imbalance

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