1887

Abstract

Summary: An NADP-dependent constitutive alcohol dehydrogenase that can oxidize hexan-1-ol was detected in several Gram-positive and Gram-negative eubacteria and in two yeasts. The enzyme was purified to homogeneity from NCIB 8250 and from D273–10B. The bacterial enzyme appears to be a tetramer of subunit 40 300 and the yeast enzyme appears to be a monomer of subunit 43 500. The N-terminal amino acid sequence of the bacterial enzyme has 34% identity with part of the sequence of a fermentative alcohol dehydrogenase from The pl value of the bacterial enzyme was 5.7 and the pH optimum was 10.2. Both the bacterial and yeast enzymes were shown to transfer the -R hydrogen to/from NADP(H). The substrate specificities of the two enzymes were similar to each other, both oxidizing primary alcohols and some diols, but not secondary alcohols. The maximum velocities of both enzymes were with pentan-1-ol as substrate and there was very low activity with ethanol; the maximum specificity constants were found with primary alcohols containing six to eight carbon atoms. Neither enzyme was significantly inhibited by metal-binding agents but some thiol-blocking compounds inhibited them. It appears that these two alcohol dehydrogenases, one prokaryotic and one eukaryotic, are structurally, kinetically and functionally different from members of the major known groups of alcohol dehydrogenases.

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1994-01-01
2024-03-29
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References

  1. Allison N., O'Donnell M. J., Hoey M. E., Fewson C. A. Membrane-bound lactate dehydrogenases and mandelate dehydrogenases of Acinetobacter calcoaceticus Purification and properties. Biochem J 1985; 231:407–416
    [Google Scholar]
  2. Arnold L. J. Jr, You K., Allison W. S., Kaplan N. O. Determination of the hydride transfer stereospecificity of nicotinamide adenine dinucleotide linked oxidoreduction by proton magnetic resonance. Biochemistry 1976; 15:4844–4849
    [Google Scholar]
  3. Asperger O., Kleber H.-P. Metabolism of alkanes by Acinetobacter. In The Biology of Acinetobacter 1991 Towner K. J., Bergogne-Berezin E., Fewson C. A. Edited by New York and London: Plenum Press; pp. 323–350.
    [Google Scholar]
  4. Baker M. E. Sequence similarity between Pseudomonas dihydrodiol dehydrogenase, part of the gene cluster that metabolizes polychlorinated biphenyls, and dehydrogenases involved in the metabolism of ribitol and glucitol and synthesis of antibiotics and 17/i-oestradiol, testosterone and corticosterone. Biochem J 1990; 267:839–841
    [Google Scholar]
  5. Baker D. P., Fewson C. A. Purification and characterization of d( - )-mandelate dehydrogenase from Rhodotorula graminis. J Gen Microbiol 1989; 135:2035–2044
    [Google Scholar]
  6. Baker D. P., Kleanthous C., Keen J. N., Fewson C. A. Mechanistic and active site studies on D( -)-mandelate dehydrogenase from Rhodotorula graminis. Biochem J 1992; 281:211–218
    [Google Scholar]
  7. Blakely R. L., Ramasastri B. V., McDougall B. M. The biosynthesis of thymidylic acid V. Hydrogen isotope studies with dihydrofolate reductase and thymidylate synthetase. J Biol Chem 1963; 238:3075–3079
    [Google Scholar]
  8. Brändén C.-I., Jörnvall H., Eklund H., Furugren B. Alcohol dehydrogenases. In The Enzymes 1975, 3rd edn. vol. XI pp. Edited by Boyer P. D. London: Academic Press;103–190
    [Google Scholar]
  9. Clark D. P. Evolution of bacterial alcohol metabolism. In The Evolution of Metabolic Function 1992 pp. Edited by Mortlock R. P., Raton Boca, Arbor Ann. London: CRC Press;105–114
    [Google Scholar]
  10. DeBruyn J., Johannes A., Weckx M., Beumer-Johannes M.-P. Partial purification and characterization of a NADP- dependent broad specificity alcohol dehydrogenase from Mycobacterium tuberculosis var bovis. J Gen Microbiol 1981; 124:359–363
    [Google Scholar]
  11. Eisenthal R., Cornish-Bowden A. The direct linear plot. Biochem J 1974; 139:715–720
    [Google Scholar]
  12. Fersht A. Enzyme Structure and Mechanism 1985 pp. New York: W. H. Freeman;392–397
    [Google Scholar]
  13. Findlay J. B. C., Pappin D. J. C., Keen J. N. Automated solid-phase microsequencing. In Protein Sequencing. A Practical Approach 1989 pp. Edited by Findlay J. B. C., Geisgow M. Oxford: IRL Press;69–84
    [Google Scholar]
  14. Fixter L. M., Nagi M. N. The presence of an NADP- dependent alcohol dehydrogenase activity in Acinetobacter calcoaceticus. FEAIS Microbiol Eett 1984; 22:297–299
    [Google Scholar]
  15. Fox M. G. A., Dickinson F. M., Ratledge C. Long-chain alcohol and aldehyde dehydrogenases in Acinetobacter calcoaceticus strain HOl-N. J Gen Microbiol 1992; 138:1963–1972
    [Google Scholar]
  16. Glasfeld A., Benner S. A. The stereospecificity of the ferrous-ion-dependent alcohol dehydrogenase from Zymomonas mobilis. Eur J Biochem 1989; 180:373–375
    [Google Scholar]
  17. Goodlove P. E., Cunningham P. R., Parker J., Clark D. P. Cloning and sequence analysis of the fermentative alcohol- dehydrogenase-encoding gene of Escherichia coli. Gene 1989; 85:209–214
    [Google Scholar]
  18. Hobbs G., Frazer C. M., Gardner D. C. J., Cullum J. A., Oliver S. G. Dispersed growth of Streptomyces in liquid culture. Appl Microbiol Biotechnol 1989; 31:272–277
    [Google Scholar]
  19. Hodgman C. D., Weast R. C., Selby S. M. (editors) Handbook of Chemistry&Physics 1959 Cleveland: Chemical Rubber Publishing;
    [Google Scholar]
  20. Holms W. H., Bennett P. M. Regulation of isocitrate dehydrogenase activity in Escherichia coli on adaptation to acetate. J Gen Microbiol 1971; 65:57–68
    [Google Scholar]
  21. Hopwood D. A., Bibb M. J., Chater K. F., Kieser K., Bruton C. J., Kieser H. M., Lydiate D. J., Smith C. P., Ward J. M., Schrempf H. Genetic Manipulation of Streptomyces: a Eabora- tory Manual; 1985 p. Norwich: John Innes Foundation;239
    [Google Scholar]
  22. Jörnvall H., Persson M., Jeffery J. Alcohol and polyol dehydrogenases are both divided into two protein types, and structural properties cross-relate the different enzyme activities within each type; 1981 Proc Natl Acad Sci USA: 784226–4230
    [Google Scholar]
  23. Jörnvall H., Persson B., Jeffery J. Characterization of alcohol/polyol dehydrogenases (zinc-containing long-chain alcohol dehydrogenases). Biochem J 1987; 167:195–201
    [Google Scholar]
  24. Koivusalo M., Uotila L. Glutathione-dependent formaldehyde dehydrogenase: evidence for the identity with class III alcohol dehydrogenase. In Engymology and Molecular Biology of Carbonyl Metabolism 1991 vol. 3 pp. Edited by Weiner H., Wermuth B., Crabb D. W. New York&London: Plenum Press;337–345
    [Google Scholar]
  25. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685
    [Google Scholar]
  26. MacKintosh R. W., Fewson C. A. Microbial aromatic alcohol and aldehyde dehydrogenases. In Engymology and Molecular Biology of Carbonyl Metabolism 1987 pp. Edited by Weiner H., Flynn T. G. New York: Alan R. Liss;259–273
    [Google Scholar]
  27. MacKintosh R. W., Fewson C. A. Benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase from Acinetobacter calcoaceticus. Purification and properties. Biochem J 1988; 250:743–751
    [Google Scholar]
  28. Murray K., Dugglesby C. J., Sala-Trepat J. M., Williams P. A. The metabolism of benzoate and methylbenzoates via the meta cleavage pathway. Eur J Biochem 1972; 29:301–310
    [Google Scholar]
  29. Pocker Y., Li H. Kinetics and mechanism of methanol and formaldehyde interconversion and formaldehyde oxidation catalysed by liver alcohol dehydrogenase. In Engymology and Molecular Biology of Carbonyl Metabolism 1991 vol. 3 pp. Edited by Weiner H., Wermuth B., Crabb D. W. New York&London: Plenum Press;337–345
    [Google Scholar]
  30. Reid G. A., Schatz G. Import of proteins into mitochondria. J Biol Chem 1982; 257:13056–13061
    [Google Scholar]
  31. Tassin J.-P., Vandecasteele J.-P. Etude d’une alcool deshydrogenase utilisant les alcools a longue chaine. C R Acad Sci Pans Se D 1971; s272:1024–1026
    [Google Scholar]
  32. Tassin J.-P., Vandecasteele J.-P. Separation and characterization of long-chain alcohol dehydrogenase isoenzymes from Pseudomonas aeruginosa. Biochim Biophys Acta 1972; 276:31–42
    [Google Scholar]
  33. Tauchert H., Grunow M., Harnisch H., Aurich H. Purification and some properties of NADP-dependent alcohol dehydrogenase from Acinetobacter calcoaceticus. Acta Biol Med Ger 1976; 35:1267–1273
    [Google Scholar]
  34. Thatcher D. R., Sawyer L.-P. Secondary structure prediction from the amino acid sequence of Drosophila melanogaster (fruit fly) alcohol dehydrogenase. Biochem J 1980; 187:875–886
    [Google Scholar]
  35. Wales M. R., Fewson C. A. Comparison of the primary structures of bacterial alcohol dehydrogenases. In Engymology and Molecular Biology of Carbonyl Metabolism 1991 vol. 3 pp. Edited by >Weiner H., Wermuth B., Crabb D. W. New York&London: Plenum Press;337–345
    [Google Scholar]
  36. Williams P. A. Enzpack 1985 Cambridge: Elsevier-BIOSOFT;
    [Google Scholar]
  37. Williamson V. M., Paquin C. E. Homology of Saccharomyces cerevisiae ADH4 to an iron-activated alcohol dehydrogenase from Zymomonas mobilis. Mol&Gen Genet 1987; 209:374–381
    [Google Scholar]
  38. Young E. T., Pilgrim D. Isolation and DNA sequence of ADH3, a nuclear gene encoding the mitochondrial isozyme of alcohol dehydrogenase in Saccharomyces cerevisiae. Mol Cell Biol 1985; 5:3025–3034
    [Google Scholar]
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