Ruyter G J, Postma P W, van Dam K
E. C. Slater Institute for Biochemical Research, University of Amsterdam, The Netherlands.
J Bacteriol. 1991 Oct;173(19):6184-91. doi: 10.1128/jb.173.19.6184-6191.1991.
The quantitative effects of variations in the amount of enzyme IIGlc of the phosphoenolpyruvate:glucose phosphotransferase system (PTS) on glucose metabolism in Escherichia coli were studied. The level of enzyme IIGlc could be adjusted in vivo to between 20 and 600% of the wild-type chromosomal level by using the expression vector pTSG11. On this plasmid, expression of the structural gene for enzyme IIGlc, ptsG, is controlled by the tac promoter. As expected, the control coefficient (i.e., the relative increase in pathway flux, divided by the relative increase in amount of enzyme) of enzyme IIGlc decreased in magnitude if a more extensive pathway was considered. Thus, at the wild-type level of enzyme IIGlc activity, the control coefficient of this enzyme on the growth rate on glucose and on the rate of glucose oxidation was low, while the control coefficient on uptake and phosphorylation of methyl alpha-glucopyranoside (an enzyme IIGlc-specific, nonmetabolizable glucose analog) was relatively high (0.55 to 0.65). The implications of our findings for PTS-mediated regulation, i.e., inhibition of growth on non-PTS compounds by glucose, are discussed.
葡萄糖磷酸转移酶系统(PTS)中IIGlc酶量的变化对大肠杆菌葡萄糖代谢的定量影响。通过使用表达载体pTSG11,可在体内将IIGlc酶水平调节至野生型染色体水平的20%至600%之间。在该质粒上,IIGlc酶的结构基因ptsG的表达由tac启动子控制。正如预期的那样,如果考虑更广泛的途径,IIGlc酶的控制系数(即途径通量的相对增加量除以酶量的相对增加量)会减小。因此,在IIGlc酶活性的野生型水平下,该酶对葡萄糖生长速率和葡萄糖氧化速率的控制系数较低,而对α-甲基葡萄糖苷(一种IIGlc酶特异性、不可代谢的葡萄糖类似物)的摄取和磷酸化的控制系数相对较高(0.55至0.65)。讨论了我们的发现对PTS介导的调节的意义,即葡萄糖对非PTS化合物生长的抑制作用。