Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. 1991 Oct; 173(19): 5975–5982.
PMCID: PMC208341

Gel electrophoretic analysis of Zymomonas mobilis glycolytic and fermentative enzymes: identification of alcohol dehydrogenase II as a stress protein.


The 13 major enzymes which compose the glycolytic and fermentative pathways in Zymomonas mobilis are particularly abundant and represent one-half of the soluble protein in exponential-phase cells. One- and two-dimensional polyacrylamide gel electrophoresis maps were developed for 12 of these enzymes. Assignments were made by comigration with purified proteins, comparison with overexpressed genes in recombinant strains, and Western blots (immunoblots). Although most glycolytic enzymes appeared resistant to turnover and accumulated in stationary-phase cells, the protein levels of pyruvate kinase, alcohol dehydrogenase I, and glucokinase declined. Alcohol dehydrogenase II was identified as a major stress protein and was induced both by exposure to ethanol and by elevated temperature (45 degrees C). This enzyme, encoded by the adhB gene, is expressed from tandem promoters which share partial sequence identity with the Escherichia coli consensus sequence for heat shock proteins.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.7M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barnell WO, Yi KC, Conway T. Sequence and genetic organization of a Zymomonas mobilis gene cluster that encodes several enzymes of glucose metabolism. J Bacteriol. 1990 Dec;172(12):7227–7240. [PMC free article] [PubMed]
  • Blake MS, Johnston KH, Russell-Jones GJ, Gotschlich EC. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. [PubMed]
  • Conway T, Byun MO, Ingram LO. Expression Vector for Zymomonas mobilis. Appl Environ Microbiol. 1987 Feb;53(2):235–241. [PMC free article] [PubMed]
  • Conway T, Ingram LO. Phosphoglycerate kinase gene from Zymomonas mobilis: cloning, sequencing, and localization within the gap operon. J Bacteriol. 1988 Apr;170(4):1926–1933. [PMC free article] [PubMed]
  • Conway T, Osman YA, Konnan JI, Hoffmann EM, Ingram LO. Promoter and nucleotide sequences of the Zymomonas mobilis pyruvate decarboxylase. J Bacteriol. 1987 Mar;169(3):949–954. [PMC free article] [PubMed]
  • Conway T, Sewell GW, Ingram LO. Glyceraldehyde-3-phosphate dehydrogenase gene from Zymomonas mobilis: cloning, sequencing, and identification of promoter region. J Bacteriol. 1987 Dec;169(12):5653–5662. [PMC free article] [PubMed]
  • Conway T, Sewell GW, Osman YA, Ingram LO. Cloning and sequencing of the alcohol dehydrogenase II gene from Zymomonas mobilis. J Bacteriol. 1987 Jun;169(6):2591–2597. [PMC free article] [PubMed]
  • Eddy CK, Mejia JP, Conway T, Ingram LO. Differential expression of gap and pgk genes within the gap operon of Zymomonas mobilis. J Bacteriol. 1989 Dec;171(12):6549–6554. [PMC free article] [PubMed]
  • Jablonski PE, DiMarco AA, Bobik TA, Cabell MC, Ferry JG. Protein content and enzyme activities in methanol- and acetate-grown Methanosarcina thermophila. J Bacteriol. 1990 Mar;172(3):1271–1275. [PMC free article] [PubMed]
  • Keshav KF, Yomano LP, An HJ, Ingram LO. Cloning of the Zymomonas mobilis structural gene encoding alcohol dehydrogenase I (adhA): sequence comparison and expression in Escherichia coli. J Bacteriol. 1990 May;172(5):2491–2497. [PMC free article] [PubMed]
  • Kroh HE, Simon LD. The ClpP component of Clp protease is the sigma 32-dependent heat shock protein F21.5. J Bacteriol. 1990 Oct;172(10):6026–6034. [PMC free article] [PubMed]
  • Kusukawa N, Yura T, Ueguchi C, Akiyama Y, Ito K. Effects of mutations in heat-shock genes groES and groEL on protein export in Escherichia coli. EMBO J. 1989 Nov;8(11):3517–3521. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lindquist S, Craig EA. The heat-shock proteins. Annu Rev Genet. 1988;22:631–677. [PubMed]
  • Mackenzie KF, Conway T, Aldrich HC, Ingram LO. 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. [PMC free article] [PubMed]
  • Mierendorf RC, Percy C, Young RA. Gene isolation by screening lambda gt11 libraries with antibodies. Methods Enzymol. 1987;152:458–469. [PubMed]
  • Merril CR, Goldman D, Van Keuren ML. Gel protein stains: silver stain. Methods Enzymol. 1984;104:441–447. [PubMed]
  • Michel GP, Starka J. Effect of ethanol and heat stresses on the protein pattern of Zymomonas mobilis. J Bacteriol. 1986 Mar;165(3):1040–1042. [PMC free article] [PubMed]
  • Neale AD, Scopes RK, Kelly JM, Wettenhall RE. The two alcohol dehydrogenases of Zymomonas mobilis. Purification by differential dye ligand chromatography, molecular characterisation and physiological roles. Eur J Biochem. 1986 Jan 2;154(1):119–124. [PubMed]
  • Neale AD, Scopes RK, Wettenhall RE, Hoogenraad NJ. Pyruvate decarboxylase of Zymomonas mobilis: isolation, properties, and genetic expression in Escherichia coli. J Bacteriol. 1987 Mar;169(3):1024–1028. [PMC free article] [PubMed]
  • Neale AD, Scopes RK, Wettenhall RE, Hoogenraad NJ. Nucleotide sequence of the pyruvate decarboxylase gene from Zymomonas mobilis. Nucleic Acids Res. 1987 Feb 25;15(4):1753–1761. [PMC free article] [PubMed]
  • O'Farrell PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed]
  • Osman YA, Conway T, Bonetti SJ, Ingram LO. Glycolytic flux in Zymomonas mobilis: enzyme and metabolite levels during batch fermentation. J Bacteriol. 1987 Aug;169(8):3726–3736. [PMC free article] [PubMed]
  • Pawluk A, Scopes RK, Griffiths-Smith K. Isolation and properties of the glycolytic enzymes from Zymomonas mobilis. The five enzymes from glyceraldehyde-3-phosphate dehydrogenase through to pyruvate kinase. Biochem J. 1986 Aug 15;238(1):275–281. [PMC free article] [PubMed]
  • Scopes RK. An iron-activated alcohol dehydrogenase. FEBS Lett. 1983 Jun 13;156(2):303–306. [PubMed]
  • Scopes RK. Use of differential dye-ligand chromatography with affinity elution for enzyme purification: 2-keto-3-deoxy-6-phosphogluconate aldolase from Zymomonas mobilis. Anal Biochem. 1984 Feb;136(2):525–529. [PubMed]
  • Scopes RK, Griffiths-Smith K. Use of differential dye-ligand chromatography with affinity elution for enzyme purification: 6-phosphogluconate dehydratase from Zymomonas mobilis. Anal Biochem. 1984 Feb;136(2):530–534. [PubMed]
  • Scopes RK, Testolin V, Stoter A, Griffiths-Smith K, Algar EM. Simultaneous purification and characterization of glucokinase, fructokinase and glucose-6-phosphate dehydrogenase from Zymomonas mobilis. Biochem J. 1985 Jun 15;228(3):627–634. [PMC free article] [PubMed]
  • Spence J, Cegielska A, Georgopoulos C. Role of Escherichia coli heat shock proteins DnaK and HtpG (C62.5) in response to nutritional deprivation. J Bacteriol. 1990 Dec;172(12):7157–7166. [PMC free article] [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PMC free article] [PubMed]
  • White MF, Fothergill-Gilmore LA. Sequence of the gene encoding phosphoglycerate mutase from Saccharomyces cerevisiae. FEBS Lett. 1988 Mar 14;229(2):383–387. [PubMed]
  • Wills C, Kratofil P, Londo D, Martin T. Characterization of the two alcohol dehydrogenases of Zymomonas mobilis. Arch Biochem Biophys. 1981 Sep;210(2):775–785. [PubMed]
  • Wold F. In vivo chemical modification of proteins (post-translational modification). Annu Rev Biochem. 1981;50:783–814. [PubMed]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...