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. 1985 Dec;164(3):1324–1331. doi: 10.1128/jb.164.3.1324-1331.1985

Differential expression of hydrogenase isoenzymes in Escherichia coli K-12: evidence for a third isoenzyme.

R G Sawers, S P Ballantine, D H Boxer
PMCID: PMC219333  PMID: 3905769

Abstract

The cellular contents of the nickel-containing, membrane-bound hydrogenase isoenzymes 1 and 2 (hydrogenases 1 and 2) were analyzed by crossed immunoelectrophoresis. Their expression was differentially influenced by nutritional and genetic factors. Hydrogenase 2 content was enhanced after growth with either hydrogen and fumarate or glycerol and fumarate and correlated reasonably with cellular hydrogen uptake capacity. Hydrogenase 1 content was negligible under the above conditions but was enhanced by exogenous formate. Its expression was greatly reduced in a pfl mutant, which is unable to synthesise formate, but was restored to normal levels when the growth medium included formate. A mutation in the anaerobic regulatory gene, fnr, led to low overall hydrogenase activity and greatly reduced levels of both isoenzymes and abolished the formate enhancement of hydrogenase 1 content. Formate hydrogenlyase activity was similarly reduced in the fnr strain but, in contrast, was restored, as was overall hydrogenase activity, to normal levels by growth in the presence of formate. Low H2 uptake activity was found for the fnr strain under all growth conditions examined. Hydrogenase 1 content, therefore, does not correlate with formate hydrogenlyase activity and its role is unclear. A third hydrogenase isoenzyme, immunologically distinct from hydrogenases 1 and 2, whose expression is enhanced by formate, is present and forms part of the formate hydrogenlyase. We suggest that the effect of the fnr gene product on formate hydrogenlyase expression is mediated via internal formate.

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  1. Adams M. W., Mortenson L. E., Chen J. S. Hydrogenase. Biochim Biophys Acta. 1980 Dec;594(2-3):105–176. doi: 10.1016/0304-4173(80)90007-5. [DOI] [PubMed] [Google Scholar]
  2. Ballantine S. P., Boxer D. H. Nickel-containing hydrogenase isoenzymes from anaerobically grown Escherichia coli K-12. J Bacteriol. 1985 Aug;163(2):454–459. doi: 10.1128/jb.163.2.454-459.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. COHEN G. N., RICKENBERG H. V. Concentration spécifique réversible des amino acides chez Escherichia coli. Ann Inst Pasteur (Paris) 1956 Nov;91(5):693–720. [PubMed] [Google Scholar]
  4. Cole J. A. Microbial gas metabolism. Adv Microb Physiol. 1976;14(11):1–92. doi: 10.1016/s0065-2911(08)60226-x. [DOI] [PubMed] [Google Scholar]
  5. Dickie P., Weiner J. H. Purification and characterization of membrane-bound fumarate reductase from anaerobically grown Escherichia coli. Can J Biochem. 1979 Jun;57(6):813–821. doi: 10.1139/o79-101. [DOI] [PubMed] [Google Scholar]
  6. Graham A., Boxer D. H., Haddock B. A., Mandrand-Berthelot A. M., Jones R. W. Immunochemical analysis of the membrane-bound hydrogenase of Escherichia coli. FEBS Lett. 1980 May 5;113(2):167–172. doi: 10.1016/0014-5793(80)80584-9. [DOI] [PubMed] [Google Scholar]
  7. Graham A., Boxer D. H. The organization of formate dehydrogenase in the cytoplasmic membrane of Escherichia coli. Biochem J. 1981 Jun 1;195(3):627–637. doi: 10.1042/bj1950627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jones R. W. The role of the membrane-bound hydrogenase in the energy-conserving oxidation of molecular hydrogen by Escherichia coli. Biochem J. 1980 May 15;188(2):345–350. doi: 10.1042/bj1880345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Knappe J., Schacht J., Möckel W., Höpner T., Vetter H., Jr, Edenharder R. Pyruvate formate-lyase reaction in Escherichia coli. The enzymatic system converting an inactive form of the lyase into the catalytically active enzyme. Eur J Biochem. 1969 Dec;11(2):316–327. doi: 10.1111/j.1432-1033.1969.tb00775.x. [DOI] [PubMed] [Google Scholar]
  10. Krasna A. I. Regulation of hydrogenase activity in enterobacteria. J Bacteriol. 1980 Dec;144(3):1094–1097. doi: 10.1128/jb.144.3.1094-1097.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kuritzkes D. R., Zhang X. Y., Lin E. C. Use of phi(glp-lac) in studies of respiratory regulation of the Escherichia coli anaerobic sn-glycerol-3-phosphate dehydrogenase genes (glpAB). J Bacteriol. 1984 Feb;157(2):591–598. doi: 10.1128/jb.157.2.591-598.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Lambden P. R., Guest J. R. Mutants of Escherichia coli K12 unable to use fumarate as an anaerobic electron acceptor. J Gen Microbiol. 1976 Dec;97(2):145–160. doi: 10.1099/00221287-97-2-145. [DOI] [PubMed] [Google Scholar]
  14. Macy J., Kulla H., Gottschalk G. H2-dependent anaerobic growth of Escherichia coli on L-malate: succinate formation. J Bacteriol. 1976 Feb;125(2):423–428. doi: 10.1128/jb.125.2.423-428.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. PECK H. D., Jr, GEST H. Formic dehydrogenase and the hydrogenlyase enzyme complex in coli-aerogenes bacteria. J Bacteriol. 1957 Jun;73(6):706–721. doi: 10.1128/jb.73.6.706-721.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. PINSKY M. J., STOKES J. L. Requirements for formic hydrogenlyase adaptation in nonproliferating suspensions of escherichia coli. J Bacteriol. 1952 Aug;64(2):151–161. doi: 10.1128/jb.64.2.151-161.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pascal M. C., Chippaux M., Abou-Jaoudé A., Blaschkowski H. P., Knappe J. Mutants of Escherichia coli K12 with defects in anaerobic pyruvate metabolism. J Gen Microbiol. 1981 May;124(1):35–42. doi: 10.1099/00221287-124-1-35. [DOI] [PubMed] [Google Scholar]
  18. Unden G., Guest J. R. Cyclic AMP and anaerobic gene expression in E. coli. FEBS Lett. 1984 May 21;170(2):321–325. doi: 10.1016/0014-5793(84)81336-8. [DOI] [PubMed] [Google Scholar]
  19. Unden G., Guest J. R. Isolation and characterization of the Fnr protein, the transcriptional regulator of anaerobic electron transport in Escherichia coli. Eur J Biochem. 1985 Jan 2;146(1):193–199. doi: 10.1111/j.1432-1033.1985.tb08638.x. [DOI] [PubMed] [Google Scholar]
  20. Varenne S., Casse F., Chippaux M., Pascal M. C. A mutant of Escherichia coli deficient in pyruvate formate lyase. Mol Gen Genet. 1975 Nov 24;141(2):181–184. doi: 10.1007/BF00267683. [DOI] [PubMed] [Google Scholar]
  21. Yamamoto I., Ishimoto M. Hydrogen-dependent growth of Escherichia coli in anaerobic respiration and the presence of hydrogenases with different functions. J Biochem. 1978 Sep;84(3):673–679. doi: 10.1093/oxfordjournals.jbchem.a132172. [DOI] [PubMed] [Google Scholar]

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