1887

Abstract

The membrane-bound aldehyde dehydrogenase from ‘’ CCM 1774, solubilized from the membrane fraction by treatment with surfactants and subsequently purified to homogeneity, was characterized with respect to the of the dimeric enzyme (145000), pH optimum (5·1), pI value (5·3), substrate specificity towards straight-chain aldehydes, substrate inhibition and the effects of various inhibitors and ions. Methanol extracts of the membrane fraction and of the purified enzyme showed properties identical with pyrroloquinoline quinone as revealed by spectrophotometric methods and reactivity with apoquinoprotein glucose dehydrogenase. Pyrroloquinoline quinone could not be liberated by EDTA treatment. The enzyme activity of an apoenzyme could be partly restored by addition of membrane extracts containing pyrroloquinoline quinone adducts. The solubilized, purified quinoprotein aldehyde dehydrogenase and the membrane-bound enzyme differed in substrate binding, substrate inhibition, pH optimum and linkage to a -type cytochrome which acts as electron acceptor in membrane fractions. Since double-reciprocal plots of initial reaction rates with various concentrations of aldehyde or electron acceptor showed intersecting lines, and different inhibition patterns were obtained for the two forms of the enzyme, a ping-pong kinetic behaviour with the two reactants was excluded. The kinetic mechanism was suggested to be changed after solubilization from a random-order type to a compulsory-order type, and is thus atypical for quinoproteins reported so far.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-9-1705
1990-09-01
2022-01-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/9/mic-136-9-1705.html?itemId=/content/journal/micro/10.1099/00221287-136-9-1705&mimeType=html&fmt=ahah

References

  1. Adachi O., Tayama K., Shinagawa E., Matsushita K., Ameyama M. 1980; Purification and characterization of membrane-bound aldehyde dehydrogenase from Gluconobacter suboxydans.. Agricultural and Biological Chemistry 44:503–515
    [Google Scholar]
  2. Alberty R. A., Massey V. 1954; Interpretation of the pH variation of the maximal initial velocity of an enzyme catalyzed reaction. Biochimica et Biophysica Acta 13:347–353
    [Google Scholar]
  3. Ameyama M., Adachi O. 1982; Aldehyde dehydrogenase from acetic acid bacteria, membrane-bound. Methods in Enzymology 89:491–497
    [Google Scholar]
  4. Ameyama M., Matsushita K., Ohno Y., Shinagawa E., Adachi O. 1981; Existence of a novel prosthetic group, PQQ, in membrane- bound, electron transport chain-linked, primary dehydrogenases of oxidative bacteria. FEBS Letters 130:179–183
    [Google Scholar]
  5. Ameyama M., Matsushita K., Shinagawa E., Adachi O. 1987; Sugar-oxidizing respiratory chain of Gluconobacter suboxydans.Evidence for a branched respiratory chain and characterization of respiratory chain-linked cytochromes. Agricultural and Biological Chemistry 51:2943–2950
    [Google Scholar]
  6. Andrews P. 1964; Estimation of the molecular weight of proteins by Sephadex gel-filtration. Biochemical Journal 91:222–233
    [Google Scholar]
  7. Anthony C., Zatman L. J. 1967; The microbial oxidation of methanol. Purification and properties of the alcohol dehydrogenase of Pseudomonas M27. Biochemical Journal 104:953–959
    [Google Scholar]
  8. Baici A. 1981; The specific velocity plot.A graphical method for determining inhibition parameters for both linear and hyperbolic enzyme inhibitors. European Journal of Biochemistry 119:9–14
    [Google Scholar]
  9. Bergmann R. 1981 Charakterisierung der membrangebundenen Aldehyddehydrogenase von Acinetobacter calcoaceticus. Untersuchungen zu Lipid-Protein-Wechselwirkungen. PhD thesis M.Luther-Universität: Halle-Wittenberg, GDR.;
    [Google Scholar]
  10. Bergmeyer H. U. 1962; Acetaldehyd.Bestimmung mit Alkohol- Dehydrogenase ausHefe. In Methoden der Enzymatischen Analyse pp. 290–292 Bergmeyer H. U. Edited by Weinheim: Verlag Chemie;
    [Google Scholar]
  11. Brocklehurst K., Carlsson J., Kierstan M.P.J., Crook E. M. 1974; Covalent chromatography by thiol-disulfide interchange. Methods in Enzymology 34:531–544
    [Google Scholar]
  12. Chou T. C., Talalay P. 1981; Generalized equations for the analysis of inhibitors of Michaelis-Menten and higher-order kinetic systems with two or more mutually exclusive and nonexclusive inhibitors. European Journal of Biochemistry 115:207–216
    [Google Scholar]
  13. Cleland W. W. 1970; Steady state kinetics. In The Enzymes 2 p. 1 Boyer P. D. Edited by New York: Academic Press;
    [Google Scholar]
  14. Cuatrecasas P., Anfinsen C. B. 1971; Affinity chromatography. Methods in Enzymology 22:345–376
    [Google Scholar]
  15. Dalziel K. 1957; Initial steady state velocities in the evolution of enzyme-coenzyme-substrate reaction mechanisms. Acta Chemica Scandinavica 11:1706–1723
    [Google Scholar]
  16. Dekker R. H., Duine J. A., Frank J.Jzn Verweil P.E.J., Westerling J. 1982; Covalent addition of H2O, enzyme substrate and activators to pyrrolo-quinolinequinone, the coenzyme of the quinoproteins. European Journal of Biochemistry 125:69–73
    [Google Scholar]
  17. Dixon M. 1953; The effect of pH on the affinities of enzymes for substrates and inhibitors. Biochemical Journal 55:161–170
    [Google Scholar]
  18. Dokter P., Frank J.Jzn Duine J. A. 1986; Purification and characterization of quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus L.M.D. 79·41. Biochemical Journal 239:163–167
    [Google Scholar]
  19. Duine J. A., Frank J.Jzn 1980; The prosthetic group of methanol dehydrogenase.Purification and some of its properties. Biochemical Journal 187:221–226
    [Google Scholar]
  20. Duine J. A., Frank J., Van Zeeland J. K. 1979; Glucose dehydrogenase from Acinetobacter calcoaceticus: a quinoprotein. FEBS Letters 108:443–446
    [Google Scholar]
  21. Duine J. A., Frank J.Jzn Jongejan A. 1986; PQQ and quinoprotein enzymes in microbial oxidations. FEMS Microbiology Reviews 32:165–178
    [Google Scholar]
  22. Duine J. A., Frank J.Jzn Jongejan A. 1987; Enzymology of quinoproteins. Advances in Enzymology and Related Areas of Molecular Biology 59:169–212
    [Google Scholar]
  23. Dully J. R., Grieve P. A. 1975; A simple technique for eliminating interference by detergents in the Lowry method of protein determination. Analytical Biochemistry 64:136–141
    [Google Scholar]
  24. Fromm H. J. 1975 Initial Rate Enzyme Kinetics. Berlin, Heidelberg & New York:: Springer.;
    [Google Scholar]
  25. Fukaya M., Tayama K., Tamaki T., Tagami H., Okumura H., Kawamura Y., Beppu T. 1989; Cloning of the membrane- bound aldehyde dehydrogenase gene of Acetobacter polyoxogenes and improvement of acetic acid production by use of the cloned gene. Applied and Environmental Microbiology 55:171–176
    [Google Scholar]
  26. Geiger O., GÖrsch H. 1986; Crystalline glucose dehydrogenase from Acinetobacter calcoaceticus. Biochemistry 25:6043–6048
    [Google Scholar]
  27. Hedrick J. L., Smith A. J. 1968; Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Archives of Biochemistry and Biophysics 126:155–164
    [Google Scholar]
  28. Hommel R., Kleber H.-P. 1984a; Oxidation of long-chain alkanes by Acetobacter rancens. Applied Microbiology and Biotechnology 19:110–113
    [Google Scholar]
  29. Hommel R., Kleber H.-P. 1984b; A pyridine nucleotide- independent aldehyde dehydrogenase involved in the alkane oxidation of ‘Acetobacter rancens’.. FEMS Microbiology Letters 23:113–116
    [Google Scholar]
  30. Hommel R., Kleber H. P. 1984c; Purification and properties of a membrane-bound aldehyde dehydrogenase involved in the oxidation of alkanes by Acetobacter rancens CCM 1774. Proceedings of the 3rd European Congress on Biotechnology1-133–1-137 Weinheim:: Verlag Chemie;
    [Google Scholar]
  31. Hommel R., Borneleit P., Kleber H.-P. 1985; Effects of detergents and phospholipids on the pyridine nucleotide-independent aldehyde dehydrogenase from membranes of Acetobacter rancens. Biomedica Biochimica Acta 9:1315–1320
    [Google Scholar]
  32. Hommel R., Kurth J., Kleber H.-P. 1988; NADP+-dependent aldehyde dehydrogenase from ‘Acetobacter rancens’ CCM 1774: Purification and properties. Journal of Basic Microbiology 28:25–33
    [Google Scholar]
  33. Matsushita K., Adachi O., Shinagawa E., Ameyama M. 1978; Isolation and characterization of outer and inner membranes from Pseudomonas aeruginosa and effect of EDTA on the membranes. Journal of Biochemistry 83:171–181
    [Google Scholar]
  34. Osborn M. J., Gander J. E., Parisi E., Carson J. 1972; Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane. Journal of Biological Chemistry 247:3962–3972
    [Google Scholar]
  35. SchÖpp W., Aurich H. 1973; Abhängigkeit der Km- und Vmax- Werte von der Kettenlange des Substrates fur die Reaktion der AlkoholdehydrogenaseausHefe. Acta Biologica et Medica Germanica 31:19–28
    [Google Scholar]
  36. Skerman V.B.D., Mcgowan V., Sneath P.H.A. 1980; Approved lists of bacterial names. International Journal of Systematic Bacteriology 30:255–420
    [Google Scholar]
  37. Sorger H., Aurich H. 1978; Mikrobielle Aldehyddehydrogenasen und ihre Bedeutung für die Assimilation aliphatischer Kohlenwas- serstoffe. Wissenschaftliche Zeitschrift der Karl-Marx-Universität Leipzig, Mathematisch-Naturwissenschaftliche Reihe 27:35–45
    [Google Scholar]
  38. Vestenberg O., Svensson H. 1966; Isoelectric fractionation, analysis, and characterization of ampholytes in natural pH gradients. IV. Resolving power in connection with separation of myoglobins. Acta Chemica Scandinavica 20:820–834
    [Google Scholar]
  39. Weber K., Osborn M. 1969; Reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide-gel electrophoresis. Journal of Biological Chemistry 244:4406–4412
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-9-1705
Loading
/content/journal/micro/10.1099/00221287-136-9-1705
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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