1887

Abstract

The pattern of ethanol and lactate formation by continuous cultures of LMG 6564 under glucose limitation is affected by culture conditions such as pH and dilution rate. NADH- and NADPH-mediated lactate dehydrogenase (LDH; EC 1.1.1.27) and alcohol (ethanol) dehydrogenase (ADH; EC 1.1.1.1 and EC 1.1.1.2) activities were measured in cell extracts from continuous cultures grown under different conditions. In conditions of high product formation, the NADH-mediated reaction was higher than the NADPH-mediated reaction for both LDH and ADH. LDH showed an absolute requirement for fructose 1,6-bisphosphate (FBP). Both NADH- and NADPH-linked LDH reactions were cytoplasmic, not sensitive to oxygen, and had a pH optimum of 6·0–6·5; the temperature optimum was 55–60 °C. The reverse reaction (lactate oxidation) could not be demonstrated. ADH activity was found in the particulate fraction of the cell lysate and was sensitive (not completely, but irreversibly) to oxygen. The temperature and pH optima were 43 °C, pH 7·0 and 45 °C, pH 8·8 for the NADH- and NADPH-mediated reactions, respectively. The production of at least two different ADHs is likely. LDH and ADH seemed to be regulated at the level of enzyme synthesis (direct correlation between the activities and the lactate and ethanol yields in the culture) with a second regulation of LDH by FBP at the reaction level.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-10-1945
1990-10-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/10/mic-136-10-1945.html?itemId=/content/journal/micro/10.1099/00221287-136-10-1945&mimeType=html&fmt=ahah

References

  1. Andersch W., Bahl H., Gottschalk G. 1983; Level of enzymes involved in acetate, butyrate, acetone and butanol formation by Clostridium acetobutylicum. European Journal of Applied Microbiology and Biotechnology 18:327–332
    [Google Scholar]
  2. Ben-Bassat A., Lamed R., Zeikus J.G. 1981; Ethanol production by thermophilic bacteria: metabolic control of end product formation in Thermoanaerobium brockii. Journal of Bacteriology 146:192–199
    [Google Scholar]
  3. Clark S.W., Bennett G.N., Rudolph F.B. 1989; Isolation and characterization of mutants ofClostridium acetobutylicumATCC 824 deficient in acetoacetyl-coenzyme A: acetate/butyrate: coenzyme A- transferase (EC2.8.3.9) and in other solvent pathway enzymes. Applied and Environmental Microbiology 55:970–976
    [Google Scholar]
  4. Clarke A.R., Evington J.R.N., Dunn C.R., Atkinson T., Holbrook J.J. 1986; The molecular pathway by which fructose 1,6-bisphosphate induces the assembly of a bacterial lactate dehydrogenase. Biochimica et Biophysica Acta 870:112–126
    [Google Scholar]
  5. Davis B.J. 1964; Disc electrophoresis. II. Method and application to human serum proteins. Annals of the New York Academy of Sciences 121:404–427
    [Google Scholar]
  6. De Vos P., Stevens P., De Ley J. 1983; Hydrogen gas production from formate and glucose by different members of the Enterobacter- iaceae. Biotechnology Letters 5:69–74
    [Google Scholar]
  7. De Vries W., Kapteyn W.M.C., Van Der Beek E.G., Stouthamer A.H. 1970; Molar growth yields and fermentation balances ofLactobacillus caseiL3 in batch cultures and in continuous cultures. Journal of General Microbiology 63:333–345
    [Google Scholar]
  8. Freier D., Gottschalk G. 1987; l(+)-Lactate dehydrogenase ofClostridium acetobutylicumis activated by fructose-1,6-bisphosphate. FEMS Microbiology Letters 43:229–233
    [Google Scholar]
  9. Garvie E.I. 1969; Lactic dehydrogenases of strains of the genus Leuconostoc. Journal of General Microbiology 58:85–94
    [Google Scholar]
  10. Garvie E.I. 1980; Bacterial lactate dehydrogenases. Microbiological Reviews 44:106–139
    [Google Scholar]
  11. Hillmer P., Gottschalk G. 1972; Particulate nature of enzymes involved in the fermentation of ethanol and acetate by Clostridium kluyveri. FEBS Letters 21:351–354
    [Google Scholar]
  12. Hillmer P., Gottschalk G. 1974; Solubilization and partial characterization of particulate dehydrogenase from Clostridium kluyveri. Biochimica et Biophysica Acta 334:12–23
    [Google Scholar]
  13. Holdeman L.V., Cato E.P., Moore W.E.C. 1977 Anaerobe Laboratory Manual Blackburg, Virginia: Southern Printing Co.;
    [Google Scholar]
  14. Hsu E.J., Ordal Z.J. 1970; Comparative metabolism of vegetative and sporulating cultures of Clostridium thermosaccharolyticum. Journal of Bacteriology 102:369–376
    [Google Scholar]
  15. Hyun H.H., Shen G.J., Zeikus J.G. 1985; Differential amylosaccharide metabolism ofClostridium thermosulfurogenesand Clostridium thermohydrosulfuricum. Journal of Bacteriology 164:1153–1161
    [Google Scholar]
  16. Jones D.T., Woods D.R. 1986; Acetone-butanol fermentation revisited. Microbiological Reviews 50:484–524
    [Google Scholar]
  17. Jungermann K., Thauer R.K., Leimenstoll G., Decker K. 1973; Function of reduced pyridine nucleotide-ferredoxin oxidore- ductases in saccharolytic Clostridia. Biochimica et Biophysica Acta 305:268–280
    [Google Scholar]
  18. Kersters K., De Ley J. 1975; Identification and grouping of bacteria by numerical analysis of their electrophoretic protein pattern. Journal of General Microbiology 87:333–342
    [Google Scholar]
  19. Lamed R., Zeikus J.G. 1980a; Glucose fermentation pathway of Thermoanaerobium brockii. Journal of Bacteriology 141:1251–1257
    [Google Scholar]
  20. Lamed R., Zeikus J.G. 1980b; Ethanol production by thermophilic bacteria: relationship between fermentation product yields of and catabolic enzyme activities inClostridium thermocellumand Thermoanaerobium brockii. Journal of Bacteriology 144:569–578
    [Google Scholar]
  21. Lamed R., Zeikus J.G. 1981; Novel NADP-linked alcohol- aldehyde/ketone oxidoreductase in thermophilic ethanologenic bacteria. Biochemical Journal 195:183–190
    [Google Scholar]
  22. Lurz R., Mayer F., Gottschalk G. 1979; Electron microscopic study on the quaternary structure of the isolated particulate alcohol- acetaldehyde dehydrogenase complex and on its identity with the polygonal bodies ofClostridium kluyveri. Archives of Microbiology 120:255–262
    [Google Scholar]
  23. Miller G.L. 1959; Use of dinitrosalicylic reagent for the determination of reducing sugars. Analytical Chemistry 31:426–428
    [Google Scholar]
  24. Mitchell W.J., Roohi M.S., Mosely M.J., Booth I.R. 1987; Regulation of carbohydrate utilization inClostridium pasteurianum. Journal of General Microbiology 133:31–36
    [Google Scholar]
  25. Petitdemange H., Cherrier C., Raval G., Gay R. 1976; Regulation of the NADH and NADPH-ferredoxin oxidoreductases in clostridia of the butyric group. Biochimica et Biophysica Acta 421:334–347
    [Google Scholar]
  26. Rao G., Mutharasan R. 1989; NADH levels and solventogenesis inClostridium acetobutylicum:new insights through culture fluorescence. Applied Microbiology and Biotechnology 30:59–60
    [Google Scholar]
  27. Thomas T.D., Ellwood D.C., Longyear V.M. 1979; Change from homo- to heterolactic fermentation byStreptococcus lactisresulting from glucose limitation in anaerobic chemostat cultures. Journal of Bacteriology 138:109–117
    [Google Scholar]
  28. Turunen M., Parkkinen E., Londesborough J., Korhola M. 1987; Distinct forms of lactate dehydrogenase purified from ethanol- and lactate-producing cells ofClostridium thermohydrosulfuricum. Journal of General Microbiology 133:2865–2873
    [Google Scholar]
  29. Vancanneyt M., De Vos P., De Ley J. 1987; Ethanol production from glucose byClostridium thermosaccharolyticumstrains: effect of pH and temperature. Biotechnology Letters 9:567–572
    [Google Scholar]
  30. Vancanneyt M., De Vos P., Marras M., De Ley J. 1990; Ethanol production in batch and continuous culture from some carbohydrates withClostridium thermosaccharolyticumLMG 6564. Systematic and Applied Microbiology. in the Press
    [Google Scholar]
  31. Wiesenborn D.P., Rudolph F.B., Papoutsakis E.T. 1989a; Phosphotransbutyrylase fromClostridium acetobutylicumATCC824 and its role in acidogenesis. Applied and Environmental Microbiology 55:317–322
    [Google Scholar]
  32. Wiesenborn D.P., Rudolph F.B., Papoutsakis E.T. 1989b; Coenzyme A transferase fromClostridium acetobutylicumATCC 824 and its role in the uptake of acids. Applied and Environmental Microbiology 55:323–329
    [Google Scholar]
  33. Wolin M.J. 1964; Fructose, 1,6-diphosphate requirement of streptococcal lactic dehydrogenase. Science 146:775–776
    [Google Scholar]
  34. Yamada T., Carlsson J. 1975; Regulation of lactate dehydrogenase and change of fermentation products in streptococci. Journal of Bacteriology 124:55–61
    [Google Scholar]
  35. Zeikus J.G. 1979; Thermophilic bacteria: ecology, physiology and technology. Enzyme and Microbial Technology 1:243–252
    [Google Scholar]
  36. Zeikus J.G., Hegge P.W., Anderson M.A. 1979; Thermoanaerobium brockiigen. nov. and sp. nov., a new chemoorganotro- phic caldoactive, anaerobic bacterium. Archives of Microbiology 122:41–48
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-10-1945
Loading
/content/journal/micro/10.1099/00221287-136-10-1945
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error