1887

Abstract

SUMMARY: Transposon mutagenesis and antibiotic enrichment were employed to isolate a mutant of Si4 designated strain M22, that had lost the ability to grow on D-mannitol and to produce the enzyme mannitol dehydrogenase (MDH). DNA flanking the transposon in the mutant strain was used as a probe for the identification and cloning of the MDH gene . A 5.5 kb RI//II fragment from Si4 was isolated and shown to complement the mutation in M22. Successful complementation required that a promoter of the vector-plasmid pRK415 be present, suggesting that the gene is part of a larger operon. Using oligonucleotides derived from the N-terminal sequence of MDH as probes was located on the complementing fragment and the gene was sequenced. The open reading frame encodes a protein of 51 404 Da with an N-terminal sequence identical to that obtained from amino acid analysis of the purified MDH. The MDH of Si4 exhibits distant similarity to the mannitol-1-phosphate dehydrogenases from and , with 28.1% and 26.3% identity, respectively. Mutant strains deficient in MtlK displayed substantial levels of sorbitol dehydrogenase activity, originally thought to be only a minor activity associated with the MDH enzyme. It is likely that we have uncovered an additional polyol dehydrogenase with activity for sorbitol. The gene can be used for overexpression of MDH in in order to obtain sufficient amounts of enzyme for further investigations and applications.

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1993-10-01
2021-10-26
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References

  1. Berezenko S., Sturgeon R.J. 1991; The enzymatic determination of D-mannitol with mannitol dehydrogenase from Agaricus bisporus.. Carbohydrate Research 216:505–509
    [Google Scholar]
  2. Birnboim C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA.. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  3. Boyer H.W., Roulland-DUSSOIX D. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli.. Journal of Molecular Biology 41:459–472
    [Google Scholar]
  4. Davis T., Yamada M., Elgort M., Saier M.H. JR 1988; Nucleotide sequence of the mannitol (mtl) operon in Eschericia coli.. Molecular Microbiology 2:405–412
    [Google Scholar]
  5. Ditta G., Stanfield S., Corbin D., Helinski D.R. 1980; Broad host range DNA cloning system for Gram negative bacteria: construction of a gene bank of Rhizobium meliloti.. Proceedings of the National Academy of Sciences of the United States of America 77:7347–7351
    [Google Scholar]
  6. Fischer R., Pogge Von Strandmann R., Hengstenberg W. 1991; Mannitol-specific phosphoenolpyruvate-dependent phosphotransferase system of Enterococcus faecalis: Molecular cloning and nucleotide sequences of the enzyme IIImtl gene and the mannitol-1- phosphate dehydrogenase gene, expression in Escherichia coli, and comparison of the gene product with similar enzymes.. Journal of Bacteriology 173:3709–3715
    [Google Scholar]
  7. Goa J. 1953; A micro Biuret method for protein determination.. Scandinavian Journal of Clinical Investigations 5:218–222
    [Google Scholar]
  8. Horwitz S.B., Kaplan N.O. 1964; Hexitol dehydrogenases of Bacillus subtilis.. Journal of Biological Chemistry 239:830–838
    [Google Scholar]
  9. Hynes M.F., Quandt J., O’connel M.P., PÜhler A. 1989; Direct selection for curing and deletion of Rhizobium plasmids using transposons carrying the Bacillus subtilis sacB gene.. Gene 78:111–120
    [Google Scholar]
  10. Jaiang W., Wu L.F., Tomich J., Saier M.H. JR Nichhaus W.G. 1991; Swissprot Data Bank (accession no. X51359)..
    [Google Scholar]
  11. JÖRNVALL H., Persson B., Jeffery J. 1987; Characteristics of alcohol/polyol dehydrogenases.. European Journal of Biochemistry 167:195–201
    [Google Scholar]
  12. Kahle C., Schneider K.H., Giffhorn F. 1992; Pentitol metabolism of Rhodobacter sphaeroides Si4: purification and char-acterization of a ribitol dehydrogenase.. Journal of General Microbiology 138:1277–1281
    [Google Scholar]
  13. Keen N.T., Tamaki S., Kobayashi D., Trollinger D. 1988; Improved broad host-range plasmids for DNA cloning in Gramnegative bacteria.. Gene 70:191–197
    [Google Scholar]
  14. Kiba N., Inoue Y., Furusawa M. 1991; Flow-injection system for the fluorimetric determination of fructose with an immobilized mannitol dehydrogenase reactor.. Analytica Chimica Acta 243:183–186
    [Google Scholar]
  15. Loviny T., Norton P.M., Hartley B.S. 1985; Ribitol dehydrogenase of Klebsiella aerogenes. Sequence of the structural gene.. Biochemical Journal 230:579–585
    [Google Scholar]
  16. Lueking D.R., Fraley R.T., Kaplan S. 1978; Intracyto- plasmatic membrane synthesis in synchronous cell populations of Rhodopseudomonas sphaeroides.. Journal of Biological Chemistry 253:451–457
    [Google Scholar]
  17. Lunn P.G., Northrop C.A., Northrop A.J. 1989; Automated enzymatic assays for the determination of intestinal permeability probes in urine. 2. Mannitol.. Clinica Chimica Acta 183:163–170
    [Google Scholar]
  18. Marmur J. 1961; A procedure for the isolation of deoxyribonucleic acids from microorganisms.. Journal of Molecular Biology 3:208–218
    [Google Scholar]
  19. Martinez G., Barker H.A., Horecker B.L. 1963; A specific mannitol dehydrogenase from Lactobacillus brevis.. Journal of Biological Chemistry 238:1598–1603
    [Google Scholar]
  20. Moore M.D., Kaplan S. 1992; Identification of intrinsic high- level resistance to rare-earth oxides and oxyanions in members of the class Proteobacteria: Characterization of tellurite, selenite, and rhodium sesquioxide reduction in Rhodobacter sphaeroides.. Journal of Bacteriology 174:1505–1514
    [Google Scholar]
  21. Neale A.D., Scopes R.K., Kelly J.M., Wettenhall R.E.H. 1983; The two alcohol dehydrogenases of Zymomonas mobilis.. European Journal of Biochemistry 154:119–124
    [Google Scholar]
  22. Neidle E., Hartnett C., Orniston N., Bairoch A., Rekik M., Harayama S. 1992; Cis-diol dehydrogenases encoded by the TOL pWWO plasmid xylL gene and the Actinetobacter calcoaceticuschromosomal benD gene are members of the short-chain alcohol dehydrogenase superfamily.. European Journal of Biochemistry 204:113–120
    [Google Scholar]
  23. Neuberger M.S., Patterson R.A., Hartley B.S. 1979; Purification and properties of Klebsiella aerogenes D-arabitol dehydrogenase.. Biochemical Journal 183:31–42
    [Google Scholar]
  24. Persson B., Jeffery J., Jornvall H. 1991a; Different segment similarities in long-chain dehydrogenases.. Biochemical and Biophysical Research Communications 177218–223
    [Google Scholar]
  25. Persson B., Krook M., Jornvall H. 1991b; Characteristics of short-chain alcohol dehydrogenases and related enzymes.. European Journal of Biochemistry 200537–543
    [Google Scholar]
  26. Prentki P., Krisch H.M. 1984; In vitro insertional mutagenesis with a selectable DNA fragment.. Gene 29:303–313
    [Google Scholar]
  27. Rode H., Giffhorn F. 1983; Adaptation of Rhodopseudomonas sphaeroides to growth on D(—)-tartrate and large-scale production of constitutive D(—)-tartrate-dehydratase during growth on DL- malate.. Applied and Environmental Microbiology 45:716–719
    [Google Scholar]
  28. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd ed.. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press;
    [Google Scholar]
  29. Schmolke M., Bornemann A., Guder W.G. 1990; Polyol determination along the rat nephron.. Biological Chemistry HoppeSeyler 371:909–916
    [Google Scholar]
  30. Schneider K.H., Giffhorn F. 1989; Purification and properties of a polyol dehydrogenase from the phototrophic bacterium.Rhodobacter sphaeroides.. European Journal of Biochemistry 184:15–19
    [Google Scholar]
  31. Schneider K.H., Giffhorn F. 1991; Sorbitol dehydrogenase from Pseudomonas sp.: Purification, characterization and application to quantitative determination of sorbitol.. Enzyme and Microbial Technology 13:332–337
    [Google Scholar]
  32. Scopes R.K. 1983; An iron-activated alcohol dehydrogenase.. FEBS Letters 156:303–306
    [Google Scholar]
  33. Shine J., Dalgarno L. 1974; The 3-terminal sequence of Escherichia coli 16s ribosomal RNA: complementary to nonsense triplets and ribosome binding sites.. Proceedings of the National Academy of Science of the United States of America 71:1342–1346
    [Google Scholar]
  34. Simon R., Preefer U., Puhler A. 1983; A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria.. Bio/Technology 1:784–791
    [Google Scholar]
  35. Suwanto A., Kaplan S. 1992; A self-transmissable, narrow-host- range endogenous plasmid of Rhodobacter sphaeroides 2.4.1: Physical structure, incompatibility determinants, origin of replication, and transfer functions.. Journal of Bacteriology 174:1124–1134
    [Google Scholar]
  36. Taylor S.S., Rigby P.W., Hartley B.S. 1974; Ribitol dehydrogenase from Klebsiella aerogenes. Purification and subunit structure.. Biochemical Journal 141:693–700
    [Google Scholar]
  37. Wierenga R., De MAEYER M.C.H., Hol W.G.J. 1985; Interaction of pyrophosphate moieties with a-helices in dinucleotide binding proteins.. Biochemistry 24:1346–1357
    [Google Scholar]
  38. Yanisch-PERRON C., Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors.. Gene 33:103–119
    [Google Scholar]
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