Control of glucose metabolism by enzyme IIGlc of the phosphoenolpyruvate-dependent phosphotransferase system in Escherichia coli

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.

[1]  S. Roseman,et al.  The bacterial phosphoenolpyruvate: glycose phosphotransferase system. , 1990, Annual review of biochemistry.

[2]  D. Holmes,et al.  A rapid boiling method for the preparation of bacterial plasmids. , 1981, Analytical biochemistry.

[3]  B. Ames,et al.  Positive selection for loss of tetracycline resistance , 1980, Journal of bacteriology.

[4]  B. Erni Glucose transport in Escherichia coli. , 1989, FEMS microbiology reviews.

[5]  H. Kornberg,et al.  Glucose transport of Escherichia coli growing in glucose-limited continuous culture. , 1979, The Biochemical journal.

[6]  P. Postma,et al.  Phosphoenolpyruvate:carbohydrate phosphotransferase system of bacteria , 1985 .

[7]  G. Peterson,et al.  A simplification of the protein assay method of Lowry et al. which is more generally applicable. , 1977, Analytical biochemistry.

[8]  E. Lin,et al.  Substrate specificity and transport properties of the glycerol facilitator of Escherichia coli , 1980, Journal of bacteriology.

[9]  D. Hartl,et al.  Metabolic flux and fitness. , 1987, Genetics.

[10]  M. Saier,et al.  Regulation of genes coding for enzyme constituents of the bacterial phosphotransferase system , 1980, Journal of bacteriology.

[11]  W. Epstein,et al.  Phosphorylation of D-glucose in Escherichia coli mutants defective in glucosephosphotransferase, mannosephosphotransferase, and glucokinase , 1975, Journal of bacteriology.

[12]  K. Jensen,et al.  Metabolic growth rate control in Escherichia coli may be a consequence of subsaturation of the macromolecular biosynthetic apparatus with substrates and catalytic components. , 1990, Microbiological reviews.

[13]  A Danchin,et al.  The ptsH, ptsI, and crr genes of the Escherichia coli phosphoenolpyruvate-dependent phosphotransferase system: a complex operon with several modes of transcription , 1988, Journal of bacteriology.

[14]  H. Westerhoff,et al.  Quantification of the contribution of various steps to the control of mitochondrial respiration. , 1982, The Journal of biological chemistry.

[15]  P. Postma,et al.  The phosphoenolpyruvate:glucose phosphotransferase system of Salmonella typhimurium. The phosphorylated form of IIIGlc. , 1986, European journal of biochemistry.

[16]  B. Erni,et al.  Glucose permease of Escherichia coli. The effect of cysteine to serine mutations on the function, stability, and regulation of transport and phosphorylation. , 1988, The Journal of biological chemistry.

[17]  S. Roseman,et al.  Sugar transport by the bacterial phosphotransferase system. Isolation and characterization of a glucose-specific phosphocarrier protein (IIIGlc) from Salmonella typhimurium. , 1982, The Journal of biological chemistry.

[18]  R. Pietri,et al.  Localization to the inner surface of the cytoplasmic membrane by immunoelectron microscopy of enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system of Escherichia coli. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Reinhart Heinrich,et al.  A linear steady-state treatment of enzymatic chains. General properties, control and effector strength. , 1974, European journal of biochemistry.

[20]  S. Roseman,et al.  Sugar transport by the bacterial phosphotransferase system. Phosphoryl transfer reactions catalyzed by enzyme I of Salmonella typhimurium. , 1982, The Journal of biological chemistry.

[21]  D E Koshland,et al.  Characterization of rate-controlling steps in vivo by use of an adjustable expression vector. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[22]  E. Waygood,et al.  The bacterial phosphotransferase system: Kinetic characterization of the glucose, mannitol, glucitol, and N‐acetylglucosamine systems , 1986, Journal of cellular biochemistry.

[23]  S. Roseman,et al.  Modified assay procedures for the phosphotransferase system in enteric bacteria. , 1979, Analytical biochemistry.

[24]  W. Arber,et al.  Transduction of chromosomal genes and episomes in Escherichia coli. , 1960, Virology.

[25]  A. Grossman,et al.  A collection of strains containing genetically linked alternating antibiotic resistance elements for genetic mapping of Escherichia coli. , 1989, Microbiological reviews.

[26]  P. Postma,et al.  Phosphoenolpyruvate:carbohydrate phosphotransferase system of bacteria. , 1985, Microbiological reviews.

[27]  P. Postma,et al.  Competition between two pathways for sugar uptake by the phosphoenolpyruvate-dependent sugar phosphotransferase system in Salmonella typhimurium. , 2005, European journal of biochemistry.

[28]  M Rutgers,et al.  Establishment of the steady state in glucose-limited chemostat cultures of Klebsiella pneumoniae. , 1987, Journal of general microbiology.

[29]  S. Roseman,et al.  Deletion Mapping of the Genes Coding for HPr and Enzyme I of the Phosphoenolpyruvate: Sugar Phosphotransferase System in Salmonella typhimurium , 1972, Journal of bacteriology.

[30]  M. Saier,et al.  Allosteric regulation of glycerol kinase by enzyme IIIglc of the phosphotransferase system in Escherichia coli and Salmonella typhimurium , 1985, Journal of bacteriology.

[31]  S. Roseman,et al.  Sugar transport. II. Characterization of constitutive membrane-bound enzymes II of the Escherichia coli phosphotransferase system. , 1971, The Journal of biological chemistry.

[32]  S. Roseman,et al.  Sugar transport by the bacterial phosphotransferase system. The glucose receptors of the Salmonella typhimurium phosphotransferase system. , 1982, The Journal of biological chemistry.

[33]  Galactose transport in Salmonella typhimurium , 1977, Journal of bacteriology.

[34]  P. Postma,et al.  Isolation of IIIGlc of the phosphoenolpyruvate-dependent glucose phosphotransferase system of Salmonella typhimurium , 1981, Journal of bacteriology.

[35]  M. Saier Protein phosphorylation and allosteric control of inducer exclusion and catabolite repression by the bacterial phosphoenolpyruvate: sugar phosphotransferase system. , 1989, Microbiological reviews.

[36]  H. Blöcker,et al.  Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. , 1986, Gene.

[37]  E. Waygood,et al.  Determination of the levels of HPr and enzyme I of the phosphoenolpyruvate-sugar phosphotransferase system in Escherichia coli and Salmonella typhimurium. , 1983, Canadian journal of biochemistry and cell biology = Revue canadienne de biochimie et biologie cellulaire.