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Indian Journal of Microbiology

, Volume 47, Issue 2, pp 126–131 | Cite as

Glucose dehydrogenase of a rhizobacterial strain of Enterobacter asburiae involved in mineral phosphate solubilization shares properties and sequence homology with other members of enterobacteriaceae

  • C. Tripura
  • P. Sudhakar Reddy
  • M. K. Reddy
  • B. Sashidhar
  • A. R. Podile
Original Article

Abstract

Glucose dehydrogenase (GDH) of Gram-negative bacteria is a membrane bound enzyme catalyzing the oxidation of glucose to gluconic acid and is involved in the solubilization of insoluble mineral phosphate complexes. A 2.4 kb glucose dehydrogenase gene (gcd) of Enterobacter asburiae sharing extensive homology to the gcd of other enterobacteriaceae members was cloned in a PCR-based directional genome walking approach and the expression confirmed in Escherichia coli YU423 on both MacConkey glucose agar and hydroxyapatite (HAP) containing media. Mineral phosphate solubilization by the cloned E. asburiae gcd was confirmed by the release of significant amount of phosphate in HAP containing liquid medium. gcd was over expressed in E. coli AT15 (gcd::cm) and the purified recombinant protein had a high affinity to glucose, and oxidized galactose and maltose with lower affinities.

The enzyme was highly sensitive to heat and EDTA, and belonged to Type I, similar to GDH of E. coli.

Keywords

Enterobacter asburiae Glucose dehydrogenase Mineral phosphate solubilization 

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References

  1. 1.
    Alexander M (1977) Introduction to Soil Microbiology. John Wiley and sons, New York, pp. 333–339Google Scholar
  2. 2.
    Ames BN (1964) Assay of inorganic phosphate, total phosphate, and phosphatases. Methods Enzymol 8:115–118CrossRefGoogle Scholar
  3. 3.
    Dokter P, Frank J, & Duine JA (1986) Purification and characterization of quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus L. M. D. 79.41. Biochem J 239:163–167PubMedGoogle Scholar
  4. 4.
    Duine JA, Frank J, & van Zeeland JK (1979) Glucose dehydrogenase from Acinetobacter calcoaceticus: a ‘quinoprotein’. FEBS letters. 108:443–446PubMedCrossRefGoogle Scholar
  5. 5.
    Duine JA (1991) Quinoproteins: enzymes containing the quinonoid cofactor pyrroloquinoline quinone, topaquinone or tryptophan-tryptophan quinine. Eur J Biochem 200:271–284PubMedCrossRefGoogle Scholar
  6. 6.
    Dully JR, & Grieve PA (1975) A simple technique for eliminating interference by detergens in the Lowry method of protein determination. Anal Biochem 64:136–141CrossRefGoogle Scholar
  7. 7.
    Erni B (1989) Glucose transport in Escherichia coli. FEMS Microbiol Rev 63:3–24CrossRefGoogle Scholar
  8. 8.
    Fliege R, Tong S, Shibata A, Nickerson KW, & Conway T (1992) The Entner-Doudoroff pathway in Escherichia coli is induced for oxidative glucose metabolism via pyrroloquinoline quinone-dependent glucose dehydrogenase. App Environ Microbiol 58:3826–3829Google Scholar
  9. 9.
    Goldstein AH (1995) Recent progress in understanding the molecular genetics and biochemistry of calcium phosphate solubilization by Gram-negative bacteria. Biol Agri Hort 12:185–193Google Scholar
  10. 10.
    Gottschalk G (1986) In Bacterial Metabolism, 2nd eds. Springer-Verlag: BerlinGoogle Scholar
  11. 11.
    Gyaneshwar P, Parekh LJ, Archana G, Poole PS, Collins MD, Hutson RA, & Kumar NG (1999) Involvement of a phosphate starvation inducible glucose dehydrogenase in soil phosphate solubilization by Enterobacter asburiae. FEMS Microbiol Lett 171:223–229CrossRefGoogle Scholar
  12. 12.
    Nautiyal CS (1999) An efficient microbiological growth medium for screening phosphate solubilizing microorganisms. FEMS Microbiol Lett 170:265–270PubMedCrossRefGoogle Scholar
  13. 13.
    Postma PW, Broekhuizen CP, & Geerse RH (1989) The role of PEP: Carbohydrate phosphotransferase system in the regulation of bacterial metabolism. FEMS Microbiol Rev 63:69–80CrossRefGoogle Scholar
  14. 14.
    Reddy MK, Nair S, & Sopory SK (2002) A new approach for efficient directional genome walking using polymerase chain reaction. Anal Biochem 306:154–158PubMedCrossRefGoogle Scholar
  15. 15.
    Sharma V, Kumar V, Archana G, & Kumar NG (2005) Substrate specificity of glucose dehydrogenase (GDH) of Enterobacter asburiae PS13 and rock phosphate solubilization with GDH substrates as C sources. Can J Microbiol 51:477–482PubMedCrossRefGoogle Scholar
  16. 16.
    Sode K, Witarto AB, Watanabe K, Noda K, Ito S, & Tsugawa W (1994) Over expression of PQQ glucose dehydrogenase in Escherichia coli under holoenzyme forming condition. Biotechnol Lett 16:1265–1268Google Scholar
  17. 17.
    Sode K, Yoshida H, Matsumura K, Kikuchi T, Watanabe M, Yasutake N, Ito S, & Sano H (1995) Elucidation of the region responsible for EDTA tolerance in PQQ glucose dehydrogenase by constructing Escherichia coli and Acinetobacter calcoaceticus chimeric enzymes. Biochem Biophys Res Commun 211:268–273PubMedCrossRefGoogle Scholar
  18. 18.
    Yamada M, Sumi K, Matsushita K, Adachi O, & Yamada Y (1993) Topological analysis of quinoprotein glucose dehydrogenase in Escherichia coli and its ubiquinone binding site. J Biol Chem 268:12812–12817PubMedGoogle Scholar
  19. 19.
    Yamada M, Elias MD, Matsushita K, Migita CT, & Adachi O. (2003) Escherichia coli PQQ-containing quinoprotein glucose dehydrogenase: its structure comparison with other quinoproteins Biochem Biophys Acta 1647:185–192PubMedGoogle Scholar

Copyright information

© Association of Microbiologists of India 2007

Authors and Affiliations

  • C. Tripura
    • 1
  • P. Sudhakar Reddy
    • 2
  • M. K. Reddy
    • 2
  • B. Sashidhar
    • 1
  • A. R. Podile
    • 1
  1. 1.Department of Plant SciencesUniversity of Hyderabad, P. O. Central UniversityHyderabadIndia
  2. 2.International Centre for Genetic Engineering and Biotechnology (ICGEB)New DelhiIndia

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