1887

Abstract

A sorbitol dehydrogenase (SDH; L-iditol: NAD 2-oxidoreductase; EC1.1.1.14) was isolated from the phototrophic bacterium Rhodobacter sphaeroides strain M22, a transposon mutant of Si4 with the transposon inserted in the mannitol dehydrogenase (MDH) gene. SDH was purified 470-fold to apparent homogeneity by ammonium sulfate precipitation, chromatography on Phenyl-Sepharose, Q-Sepharose and Matrex Gel Red-A, and by gel filtration on Superdex 200. The relative molecular mass ( ) of the native SDH was 61 000 as calculated from its Stokes’ radius (r = 3.5 nm) and sedimentation coefficient ( ,w = 4.235). SDS-PAGE resulted in one single band representing a polypeptide with a of 29000, indicating that the native protein is a dimer. The isoelectric point of SDH was determined to be pH 4-8. The enzyme was specific for NAD and catalysed the oxidation of D-glucitol (sorbitol) to D-fructose, galactitol to D-tagatose and of L-iditol. The apparent values were NAD, 0-06 mM; D-glucitol, 6-2 mM; galactitol, 1-5 mM; NADH, 0-13 mM; D-fructose, 160 mM; and D-tagatose, 13 mM. The pH-optimum of substrate oxidation was 11-0 and that of substrate reduction 6-0-7-2. It was demonstrated that SDH is expressed in the wild-type strain Si4 together with MDH during growth on D-glucitol. Forty-four amino acids of the SDH N terminus were sequenced. This sequence exhibited 45-55% identity to the N-terminal sequence of 10 enzymes belonging to the short-chain alcohol dehydrogenase family.

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1995-08-01
2021-05-09
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References

  1. Beutler H.O. 1984; d-Sorbitol.. In Methods of Enzymatic Analysis, 3rd edn,. VI pp. 321–327 Edited by Bergmeyer H. U. Weinheim: Verlag Chemie;
    [Google Scholar]
  2. Birken S., Pisano M.A. 1976; Purification and properties of a polyol dehydrogenase from Cephalosporium chrysogenus. . J Bacteriol 125:225–232
    [Google Scholar]
  3. Delong A., Calderonurrea A., Dellaporta S.L. 1993; Sex determination gene TASSELSEED2 of maize encodes a short- chain alcohol dehydrogenase required for stage-specific floral organ abortion.. Cell 74:757–768
    [Google Scholar]
  4. Dothie J.M., Giglio J.R., Moore C.B., Taylor S.S., Hartley B.S. 1985; Ribitol dehydrogenase of Klebsiella aerogenes. Sequence and properties of wild-type and mutant strains.. Biochem J 230:569–578
    [Google Scholar]
  5. Goa J. 1953; A micro Biuret method for protein determination.. Sc and J Clin Invest 5:218–222
    [Google Scholar]
  6. Hofer B., Eltis L.D., Dowling D.N., Timmis K.N. 1993; Genetic analysis of a Pseudomonas locus encoding a pathway for biphenyl/polychlorinated biphenyl degradation.. Gene 130:47–55
    [Google Scholar]
  7. Jeffery J., Jörnvall H. 1988; Sorbitol dehydrogenase.. Adv Enzymol Relat Areas Mol Biol 61:47–106
    [Google Scholar]
  8. Jeffery J., Cederlund E., Jörnvall H. 1984; Sorbitol de-hydrogenase. The primary structure of sheep-liver enzyme.. Eur J Biochem 140:7–16
    [Google Scholar]
  9. Jörnvall H., Persson B., Jeffery J. 1987; Characteristics of alcohol/polyol dehydrogenase.. Eur J Biochem 167:195–201
    [Google Scholar]
  10. Kahle C., Schneider K.H., Giffhorn F. 1992; Pentitol metabolism of Rhodobacter sphaeroides Si4: purification and characterization of a ribitol dehydrogenase.. J Gen Microbiol 138:1277–1281
    [Google Scholar]
  11. Karlsson C., Höög J.O. 1993; Zinc coordination in mammalian sorbitol dehydrogenase.. Eur J Biochem 216:103–107
    [Google Scholar]
  12. Karlsson C., Maret W., Auld D.S., Höög J.O., Jörnvall H. 1989; Variability within mammalian sorbitol dehydrogenase. The primary structure of the human fiver enzyme.. Eur J Biochem 186:543–550
    [Google Scholar]
  13. Karlsson C, Jörnvall H., Höög J.O. 1991; Sorbitol dehydrogenase : cDNA coding for the rat enzyme.. Eur J Biochem 198:761–765
    [Google Scholar]
  14. Krook M., Marekov L., Jörnvall H. 1990; Purification and structural characterization of placental NAD+-linked 15-hydroxy- prostaglandin dehydrogenase. The primary structure reveals the enzyme belongs to the short-chain alcohol dehydrogenase family.. Biochemistry 29:738–743
    [Google Scholar]
  15. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  16. Marekov L., Krook M., Jörnvall H. 1990; Prokaryotic 2013- hydroxysteroid dehydrogenase is an enzyme of the ‘short-chain, non-metallo-enzyme ’ alcohol dehydrogenase type.. FEBS Lett 266:51–54
    [Google Scholar]
  17. Maret W., Auld D.S. 1988; Purification and characterization of human fiver sorbitol dehydrogenase.. Biochemistry 27:1622–1628
    [Google Scholar]
  18. Merril C.R., Goldman D., Sedman S.A., Ebert M.H. 1981; Ultrasensitive stain for proteins in polyacrylamide gels shows regional variations in cerebrospinal fluid proteins.. Science 211:1437–1438
    [Google Scholar]
  19. Neidle E., Hartnett G, Ornston N., Bairoch A., Rekik M., Harayama S. 1992; CA-diol dehydrogenases encoded by the TOL pWWO plasmid xylL gene and the Actinetobacter calcoaceticus chromosomal benD gene are members of the short-chain alcohol dehydrogenase superfamily.. Eur J Biochem 204:113–120
    [Google Scholar]
  20. Ng K., Ye R., Wu X.G, Wong S.L. 1992; Sorbitol dehydrogenase from Bacillus subtilis - purification, characterization and gene cloning.. J Biol Chem 267:24989–24994
    [Google Scholar]
  21. Persson B., Krook M., Jörnvall H. 1991; Characteristics of short-chain alcohol dehydrogenases and related enzymes.. Eur J Biochem 200:537–543
    [Google Scholar]
  22. Pfennig N., Lippert K.D. 1966; Uber das Vitamin B 12- Bediirfnis phototropher Schwefelbakterien.. Arch Microbiol 55:245–256
    [Google Scholar]
  23. Rode H., Giffhorn F. 1983; Adaptation of Rhodopseudomonas sphaeroides to growth on d-(ᅳ)-tartrate and large-scale production of constitutive d-(ᅳ)-tartrate-dehydratase during growth on DL- malate.. Appl Environ Microbiol 45:716–719
    [Google Scholar]
  24. Sarthy A.V., Schopp C., Idler K.B. 1994; Cloning and sequence determination of the gene encoding sorbitol dehydrogenase from Saccharomyces cerevisiae. . Gene 140:121–126
    [Google Scholar]
  25. Schneider K.H., Giffhorn F. 1989; Purification and properties of a polyol dehydrogenase from the phototrophic bacterium Rhodobacter sphaeroides. . Eur J Biochem 184:15–19
    [Google Scholar]
  26. Schneider K.H., Giffhorn F. 1991; Sorbitol dehydrogenase from Pseudomonas sp.: purification, characterization and application to quantitative determination of sorbitol.. Enzyme Microb Technol 13:332–337
    [Google Scholar]
  27. Schneider K.H., Giffhorn F. 1994; Overproduction of mannitol dehydrogenase in Rhodobacter sphaeroides. . Appl Microbiol Biotechnol 41:578–583
    [Google Scholar]
  28. Schneider K.H., Giffhorn F., Kaplan S. 1993; Cloning, nucleotide sequence and characterization of the mannitol dehydrogenase gene from Rhodobacter sphaeroides. . J Gen Microbiol 139:2475–2484
    [Google Scholar]
  29. Schneider K.H., Jäkel G., Hoffmann R., Giffhorn F. 1995; Enzyme evolution in Rhodobacter sphaeroides: selection of a mutant expressing a new galactitol dehydrogenase and biochemical characterization of the enzyme.. Microbiology 141:1865–1873
    [Google Scholar]
  30. Siegel L.M., Monty K.J. 1966; Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude extract preparations of sulfite and hydroxylamine reductases.. Biochim Biophys Acta 112:346–362
    [Google Scholar]
  31. Verwoert 1.1.G.S., Verbree E.C., van der Linden K.H., Nijkamp H.J.J., Stuitje A.R. 1992; Cloning , nucleotide sequence, and expression of the Escherichia coli fabD gene, encoding malonyl coenzyme A-acyl carrier protein transacylase.. J Bacteriol 174:2851–2857
    [Google Scholar]
  32. Wierenga R.K., DeMaeyer M.C.H., Hoi W.G.J. 1985; Interaction of pyrophosphate moieties with α-helices in dinucleotide binding proteins.. Biochemistry 24:1346–1357
    [Google Scholar]
  33. Wong B., Murray J.S., Castellanos M., Croen K.D. 1993; d- Arabitol metabolism in Candida albicans: studies of the biosynthetic pathway and the gene that encodes NAD-dependent d-arabitol dehydrogenase.. J Bacteriol 175:6314–6320
    [Google Scholar]
  34. Yamamoto-Otake H., Koyama Y., Horiuchi T., Nakano E. 1991; Cloning, sequencing, and expression of the N-acyl-d- mannosamine dehydrogenase gene from Elavobacterium sp. strain 141-8 in Escherichia coli. . Appl Environ Microbiol 57:1418–1422
    [Google Scholar]
  35. Yin S.J., Vagelopoulos N., Lundquist G., Jörnvall H. 1991; Pseudomonas 3β-hydroxysteroid dehydrogenase. Primary structure and relationship to the other steroid dehydrogenases.. Eur f Biochem 197:359–365
    [Google Scholar]
  36. Yoshimato T., Hagashi H., Kanatani A., Lin X.S., Nagai H., Oyama H., Kurazono K., Tsuru D. 1991; Cloning and sequencing of the 7α-hydroxysteroid dehydrogenase gene from Escherichia coli HB101 and characterization of the expressed enzyme.. J Bacteriol 173:2173–2179
    [Google Scholar]
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