1887

Abstract

A gene for periplasmic poly(vinyl alcohol) (PVA) dehydrogenase (PVADH) was cloned, based on the N-terminal amino acid sequence of the purified PVADH from sp. 113P3 and the sequence of the gene for PVADH (, GenBank accession no. AB190288). The recombinant PVADH tagged with hexahistidine was expressed in and purified to homogeneity. The recombinant enzyme had the same characteristics as the purified enzyme from sp. strain 113P. In addition to PVA, the recombinant PVADH could oxidize glycols such as polypropylene glycols and 1,3-butane/cyclohexanediol and 2,4-pentanediol, but neither primary nor secondary alcohols. The amino acid sequence of the recombinant PVADH showed similarity with those of PVADH from sp. strain VM15C, putative PVADHs from sp. EbN1, and species (54–25 % identity), and the quinohaemoprotein alcohol dehydrogenases (QH-ADHs) from , and (25–29 % identity). PVADHs from strains 113P3 and VM15C have a conserved superbarrel domain (SD), probable PQQ-binding amino acids in the SD and a haem-binding domain (HBD) (they should be designated QH-PVADHs), but the positions of the amino acid sequences for the HBD and SD are the reverse of those of QH-ADHs. A protein structure of QH-PVADHs is proposed. Results of dot-blot hybridization and RT-PCR indicated that the three genes encoding oxidized PVA hydrolase, PVADH and cytochrome are expressed constitutively and form an operon.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.28848-0
2006-07-01
2019-10-13
Loading full text...

Full text loading...

/deliver/fulltext/micro/152/7/1941.html?itemId=/content/journal/micro/10.1099/mic.0.28848-0&mimeType=html&fmt=ahah

References

  1. Chen, Z.-W., Matsushita, K., Yamashita, T., Fujii, T., Toyama, H., Adachi, O., Bellamy, H. D. & Mathews, F. S. ( 2002; ). Structure at 1.9 Å resolution of a quinohemoprotein alcohol dehydrogenase from Pseudomonas putida HK5. Structure 10, 837–849.[CrossRef]
    [Google Scholar]
  2. Duine, J. A. & Kawai, F. ( 1998; ). Enzymes involved in the bacterial degradation of polyethylene glycols. In Biochemical Principles and Mechanisms of Biosynthesis and Biodegradation of Polymers, pp. 222–229. Edited by A. Steinbüchel. Weiheim: Wiley-VCH.
  3. Groen, B. W., Van Kleef, A. G. & Duine, J. A. ( 1986; ). Quinohaemoprotein alcohol dehydrogenase apoenzyme from Pseudomonas testosteroni. Biochem J 234, 611–615.
    [Google Scholar]
  4. Hatanaka, T., Asahi, N. & Tsuji, M. ( 1995a; ). Purification and characterization of poly(vinyl alcohol) dehydrogenase from Pseudomonas sp.113P3. Biosci Biotechnol Biochem 59, 1813–1816.[CrossRef]
    [Google Scholar]
  5. Hatanaka, T., Kawahara, T., Asahi, N. & Tsuji, M. ( 1995b; ). Effect of the structure of poly(vinyl alcohol) on the dehydrogenation reaction by poly(vinyl alcohol) dehydrogenase from Pseudomonas sp. 113P3. Biosci Biotechnol Biochem 59, 1229–1231.[CrossRef]
    [Google Scholar]
  6. Hatanaka, T., Hashimoto, T., Kawahara, T., Takami, M., Asahi, N. & Wada, R. ( 1996; ). Biodegradability of oxidized poly(vinyl alcohol). Biosci Biotechnol Biochem 60, 1861–1863.[CrossRef]
    [Google Scholar]
  7. Jongejan, A., Jongejan, J. A. & Duine, J. A. ( 1998; ). Homology model of the quinohaemoprotein alcohol dehydrogenase from Comamonas testosteroni. Prot Eng 11, 185–198.[CrossRef]
    [Google Scholar]
  8. Kawai, F. ( 1995; ). Breakdown of plastic and polymers by microorganisms. In Advances in Biochemical Engineering/Biotecnology, vol. 52, pp.151–194. Edited by A. Fichter. Heidelberg: Springer.
  9. Klomklang, W., Tani, A., Kimbara, K., Mamoto, R., Ueda, T., Shimao, M. & Kawai, F. ( 2005; ). Biochemical and molecular characterization of a periplasmic hydrolase for oxidized polyvinyl alcohol from Sphingomonas sp. strain 113P3. Microbiology 151, 1255–1262.[CrossRef]
    [Google Scholar]
  10. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.[CrossRef]
    [Google Scholar]
  11. Matsumura, S., Tomizaka, N., Toki, A. & Toshima, K. ( 1998; ). Enzymatic degradation of poly(vinyl alcohol) and its copolymer. In Biochemical Principles and Mechanisms of Biosynthesis and Biodegradation of Polymers, vol. 9, pp. 230–238. Edited by A. Steinbüchel. Weinheim: Wiley-VCH.
  12. Matsumura, S., Tomizawa, N., Toki, A., Nishikawa, K. & Toshima, K. ( 1999; ). Novel poly(vinyl alcohol)-degrading enzyme and the degradation mechanism. Macromolecules 32, 7753–7761.[CrossRef]
    [Google Scholar]
  13. Oubrie, A., Rozeboom, H. J., Kalk, K. H., Huizinga, E. G. & Dijkstra, B. W. ( 2002; ). Crystal structure of quinohemoprotein alcohol dehydrogenase from Comamonas testosteroni. J Biol Chem 277, 3727–3732.[CrossRef]
    [Google Scholar]
  14. Sambrook, J. & Russell, D. W. ( 2001; ). Molecular Cloning: a Laboratory Manual, 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  15. Shimao, M., Ninomiya, K., Kuno, O., Kato, N. & Sakazawa, C. ( 1986; ). Existence of a novel enzyme, pyrroloquinoline quinone-dependent polyvinyl alcohol dehydrogenase, in a bacterial symbiont, Pseudomonas sp. strain VM15C. Appl Environ Microbiol 51, 268–275.
    [Google Scholar]
  16. Shimao, M., Tamogami, T., Nishi, K. & Harayama, S. ( 1996; ). Cloning and characterization of the gene encoding pyrroloquinoline quinone-dependent poly(vinyl alcohol) dehydrogenase of Pseudomonas sp. strain VM15C. Biosci Biotechnol Biochem 60, 1056–1062.[CrossRef]
    [Google Scholar]
  17. Shimao, M., Tamogami, T., Kishida, S. & Harayama, S. ( 2000; ). The gene pvaB encodes oxidized polyvinyl alcohol hydrolase of Pseudomonas sp. strain VM15C and forms an operon with the polyvinyl alcohol dehydrogenase gene pvaA. Microbiology 146, 649–657.
    [Google Scholar]
  18. Stoorvogel, J., Kraayveld, D. E., Van Sluis, C. A., Jongejan, J. A., De Vries, S. & Duine, J. A. ( 1996; ). Characterization of the gene encoding quinohaemoprotein ethanol dehydrogenase of Comamonas testosteroni. Eur J Biochem 235, 690–698.[CrossRef]
    [Google Scholar]
  19. Sugiura, M., Sakaki, T., Yabusaki, Y. & Ohkawa, H. ( 1996; ). Cloning and expression in Escherichia coli and Saccharomyces cerevisiae of a novel tobacco cytochrome P-450-like cDNA. Biochim Biophys Acta 1308, 231–240.[CrossRef]
    [Google Scholar]
  20. Tachibana, S., Kuba, N., Kawai, F., Duine, J. A. & Yasuda, M. ( 2003; ). Involvement of a quinoprotein (PQQ-containing) alcohol dehydrogenase in the degradation of polypropylene glycols by the bacterium Stenotrophomonas maltophilia. FEMS Microbiol Lett 218, 345–349.[CrossRef]
    [Google Scholar]
  21. Toyama, H., Fujii, A., Matsushita, K., Shinagawa, E., Ameyama, M. & Adachi, O. ( 1995; ). Three distinct quinoprotein alcohol dehydrogenases are expressed when Pseudomonas putida is grown on different alcohols. J Bacteriol 177, 2442–2450.
    [Google Scholar]
  22. Toyama, H., Mathews, F. S., Adachi, O. & Matsushita, K. ( 2004; ). Quinohemoprotein alcohol dehydrogenases: structure, function and physiology. Arch Biochem Biophys 428, 10–21.[CrossRef]
    [Google Scholar]
  23. Zarnt, G., Schräder, T. & Andreesen, J. R. ( 1997; ). Degradation of tetrahydrofurfuryl alcohol by Ralstonia eutropha is initiated by an inducible pyrroloquinoline quinone-dependent alcohol dehydrogenase. Appl Environ Microbiol 63, 4891–4898.
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.28848-0
Loading
/content/journal/micro/10.1099/mic.0.28848-0
Loading

Data & Media loading...

vol. , part 7, pp. 1941 - 1949

Sequence alignment of type II QH-PVADHs with type II QH-ADHs. [ PDF] (25 kb)



PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error