Molecular and General Genetics MGG

, Volume 217, Issue 2–3, pp 430–436 | Cite as

Cloning, characterization and DNA sequencing of the gene encoding the Mr50000 quinoprotein glucose dehydrogenase fromAcinetobacter calcoaceticus

  • Anne-Marie Cleton-Jansen
  • Nora Goosen
  • Kees Vink
  • Pieter van de Putte
Article

Summary

Recently we described the cloning of the gene coding for a Mr 87000 glucose dehydrogenase (GDH-A) fromAcinetobacter calcoaceticus. In this report we describe the cloning of a gene coding for a second GDH (GDH-B) with a Mr of 50000 from the same organism. This gene was isolated using a 20-mer synthetic oligonucleotide, derived from the N-terminal amino acid sequence of purified GDH-B as a probe to screen a genomic bank. From the DNA sequence of thegdhB gene, a protein can be derived of Mr 52772 with a 24 amino acid signal peptide which is removed, resulting in the mature protein with a Mr 50231. In vitro transcription-translation of thegdhB clone shows the mature and the precursor protien. The derived amino acid sequence has no obvious homology with GDH-A ofA. calcoaceticus. We show that disaccharides are specific GDH-B substrates and that 2-deoxyglucose is specific for GDH-A.

Key words

Acinetobacter calcoaceticus DNA sequence Glucose dehydrogenase Pyrroloquinoline quinone Signal sequence 

References

  1. Cleton-Jansen AM, Goosen N, Wenzel TJ, Van de Putte P (1988a) Cloning of the gene encoding quinoprotein glucose dehydrogenase fromAcinetobacter calcoaceticus: evidence for the presence of a second enzyme. J Bacteriol 170:2121–2125PubMedGoogle Scholar
  2. Cleton-Jansen AM, Goosen N, Odle G, Van de Putte P (1988b) Nucleotide sequence of the gene coding for quinoprotein glucose dehydrogenase fromAcinetobacter calcoaceticus. Nucleic Acids Res 16:6228PubMedGoogle Scholar
  3. Chomczynski P, Qasba PK (1984) Alkaline transfer of DNA to plastic membrane. Biochem Biophys Res Commun 122:340–344PubMedCrossRefGoogle Scholar
  4. Deveraux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:443–446Google Scholar
  5. Dokter P, Frank J, Duine JA (1986) Purification and characterization of quinoprotein glucose dehydrogenase fromAcinetobacter calcoaceticus LMD 79.41. Biochem J 239:163–169PubMedGoogle Scholar
  6. Duine JA, Frank J, Van Zeeland K (1979) Glucose dehydrogenase fromAcinetobacter calcoaceticus: a “quinoprotein”. FEBS Lett 108:443–446PubMedCrossRefGoogle Scholar
  7. Fdman P, Begg G (1967) A protein sequenator. Eur J Biochem 1:80–91CrossRefGoogle Scholar
  8. Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6–13PubMedCrossRefGoogle Scholar
  9. Geiger O, Görisch H (1986) Crystalline quinoprotein glucose dehydrogenase fromAcinetobacter calcoaceticus. Biochemistry 25:6043–6048CrossRefGoogle Scholar
  10. Goddard JM, Caput D, Williams SR, Martin Jr DW (1983) Cloning of human purine-nucleoside phosphorylase cDNA sequences by complementation inE. coli. Proc Natl Acad Sci USA 80:4281–4285PubMedCrossRefGoogle Scholar
  11. Goosen N, Vermaas DAM, Van de Putte P (1987) Cloning of the genes involved in the synthesis of coenzyme pyrroloquinoline quinone fromAcinetobacter calcoaceticus. J Bacteriol 169:303–307PubMedGoogle Scholar
  12. Hauge JG (1966) Glucose dehydrogenase:Pseudomonas sp. andBacterium anitratum. Methods Enzymol 9:92–98CrossRefGoogle Scholar
  13. Hommes RJW, Postma PW, Neijsel OM, Tempest DW, Dokter P, Duine JA (1984) Evidence of a quinoprotein glucose dehygenase apoenzyme in several strains ofEscherichia coli. FEMS Microbiol Lett 24:329–333CrossRefGoogle Scholar
  14. Hong GF (1982) A systematic sequencing strategy. J Mol Biol 158:539–549PubMedCrossRefGoogle Scholar
  15. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  16. Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New YorkGoogle Scholar
  17. Matsushita K, Shinagawa E, Inoue T, Adachi O, Ameyama M (1986) Immunological evidence for two types of PQQ dependentd-glucose dehydrogenase in bacterial membranes and the location of the enzyme inEscherichia coli. FEMS Microbiol Lett 37:141–144Google Scholar
  18. Messing J, Crea R, Seeberg PH (1981) A system for shotgun DNA sequencing. Nucleic Acids Res 9:309–321PubMedGoogle Scholar
  19. Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-termination inhibitors. Proc Natl Acad Sci USA 74:5463–5467PubMedCrossRefGoogle Scholar
  20. Simon R, Priefer U, Puhler A (1983) A broad hostrange mobilization system for in vitro engineering: transposon mutagenis in gram-negative bacteria. Bio/Technology 1:742CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • Anne-Marie Cleton-Jansen
    • 1
  • Nora Goosen
    • 1
  • Kees Vink
    • 1
  • Pieter van de Putte
    • 1
  1. 1.Laboratory of Molecular GeneticsUniversity of LeidenLeidenThe Netherlands

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