Abstract
The competitive inhibition of fructokinase by glucose has been proposed as the mechanism by which Zymomonas mobilis preferentially consumes glucose from mixtures of glucose and fructose and accumulates fructose when growing on sucrose. In this study, incorporation of radioactive fructose into biomass was used as a measure of fructose catabolism. It was determined that the rate of fructose incorporation by Z. mobilis CP4 was somewhat lower in the presence of an equimolar concentration of glucose but that the inhibition of fructokinase by glucose was not nearly as severe in vivo as was predicted from in vitro studies. Interestingly, addition of glucose to a culture of Z. mobilis CP4-M2, a glucokinaseless mutant, resulted in an immediate and nearly complete inhibition of fructose incorporation. Furthermore, addition of nonmetabolizeable glucose analogs had a similar effect on fructose catabolism by the wild-type Z. mobilis CP4, and fructose uptake by Z. mobilis CP4-M2 was shown to be severely inhibited by equimolar amounts of glucose. These results suggest that competition for fructose transport plays an important role in preferential catabolism of glucose from sugar mixtures. Indeed, the apparent K(infm) values for sugar uptake by Z. mobilis CP4 were approximately 200 mM for fructose and 13 mM for glucose. Other experiments supported the conclusion that a single facilitated diffusion transport system, encoded by the glf gene, is solely responsible for the uptake of both glucose and fructose. The results are discussed with regard to the hypothesis that the kinetics of sugar transport and phosphorylation allow the preferential consumption of glucose and accumulation of fructose, making the fructose available for the enzyme glucose-fructose oxidoreductase, which forms sorbitol, an important osmoprotectant for Z. mobilis when growing in the presence of high sugar concentrations.
Full Text
The Full Text of this article is available as a PDF (223.9 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Barnell W. O., Liu J., Hesman T. L., O'Neill M. C., Conway T. The Zymomonas mobilis glf, zwf, edd, and glk genes form an operon: localization of the promoter and identification of a conserved sequence in the regulatory region. J Bacteriol. 1992 May;174(9):2816–2823. doi: 10.1128/jb.174.9.2816-2823.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Conway T., Byun M. O., Ingram L. O. Expression Vector for Zymomonas mobilis. Appl Environ Microbiol. 1987 Feb;53(2):235–241. doi: 10.1128/aem.53.2.235-241.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Conway T. The Entner-Doudoroff pathway: history, physiology and molecular biology. FEMS Microbiol Rev. 1992 Sep;9(1):1–27. doi: 10.1111/j.1574-6968.1992.tb05822.x. [DOI] [PubMed] [Google Scholar]
- Dimarco A. A., Romano A. H. d-Glucose Transport System of Zymomonas mobilis. Appl Environ Microbiol. 1985 Jan;49(1):151–157. doi: 10.1128/aem.49.1.151-157.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hesman T. L., Barnell W. O., Conway T. Cloning, characterization, and nucleotide sequence analysis of a Zymomonas mobilis phosphoglucose isomerase gene that is subject to carbon source-dependent regulation. J Bacteriol. 1991 May;173(10):3215–3223. doi: 10.1128/jb.173.10.3215-3223.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Loos H., Krämer R., Sahm H., Sprenger G. A. Sorbitol promotes growth of Zymomonas mobilis in environments with high concentrations of sugar: evidence for a physiological function of glucose-fructose oxidoreductase in osmoprotection. J Bacteriol. 1994 Dec;176(24):7688–7693. doi: 10.1128/jb.176.24.7688-7693.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mackenzie K. F., Conway T., Aldrich H. C., Ingram L. O. Expression of Zymomonas mobilis adhB (encoding alcohol dehydrogenase II) and adhB-lacZ operon fusions in recombinant Z. mobilis. J Bacteriol. 1989 Sep;171(9):4577–4582. doi: 10.1128/jb.171.9.4577-4582.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osman Y. A., Conway T., Bonetti S. J., Ingram L. O. Glycolytic flux in Zymomonas mobilis: enzyme and metabolite levels during batch fermentation. J Bacteriol. 1987 Aug;169(8):3726–3736. doi: 10.1128/jb.169.8.3726-3736.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parker C., Barnell W. O., Snoep J. L., Ingram L. O., Conway T. Characterization of the Zymomonas mobilis glucose facilitator gene product (glf) in recombinant Escherichia coli: examination of transport mechanism, kinetics and the role of glucokinase in glucose transport. Mol Microbiol. 1995 Mar;15(5):795–802. doi: 10.1111/j.1365-2958.1995.tb02350.x. [DOI] [PubMed] [Google Scholar]
- Scopes R. K., Testolin V., Stoter A., Griffiths-Smith K., Algar E. M. Simultaneous purification and characterization of glucokinase, fructokinase and glucose-6-phosphate dehydrogenase from Zymomonas mobilis. Biochem J. 1985 Jun 15;228(3):627–634. doi: 10.1042/bj2280627. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simons R. W., Houman F., Kleckner N. Improved single and multicopy lac-based cloning vectors for protein and operon fusions. Gene. 1987;53(1):85–96. doi: 10.1016/0378-1119(87)90095-3. [DOI] [PubMed] [Google Scholar]
- Swings J., De Ley J. The biology of Zymomonas. Bacteriol Rev. 1977 Mar;41(1):1–46. doi: 10.1128/br.41.1.1-46.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weisser P., Krämer R., Sahm H., Sprenger G. A. Functional expression of the glucose transporter of Zymomonas mobilis leads to restoration of glucose and fructose uptake in Escherichia coli mutants and provides evidence for its facilitator action. J Bacteriol. 1995 Jun;177(11):3351–3354. doi: 10.1128/jb.177.11.3351-3354.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zachariou M., Scopes R. K. Glucose-fructose oxidoreductase, a new enzyme isolated from Zymomonas mobilis that is responsible for sorbitol production. J Bacteriol. 1986 Sep;167(3):863–869. doi: 10.1128/jb.167.3.863-869.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zembrzuski B., Chilco P., Liu X. L., Liu J., Conway T., Scopes R. Cloning, sequencing, and expression of the Zymomonas mobilis fructokinase gene and structural comparison of the enzyme with other hexose kinases. J Bacteriol. 1992 Jun;174(11):3455–3460. doi: 10.1128/jb.174.11.3455-3460.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
