Purification and Characterization of meta-Cleavage Compound Hydrolase from a Carbazole Degrader Pseudomonas resinovorans Strain CA10

Hideaki Nojiri, Hiroko Taira, Kenichi Iwata, Kenichi Morii, Jeong Won Nam, Takako Yoshida, Hiroshi Habe, Shugo Nakamura, Kentaro Shimizu, Hisakazu Yamane, Toshio Omori

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15 Citations (Scopus)

Abstract

2-Hydroxy-6-oxo-6-(2′-aminophenyl)-hexa-2,4- dienoic acid [6-(2′-aminophenyl)-HODA] hydrolase, involved in carbazole degradation by Pseudomonas resinovorans strain CA10, was purified to near homogeneity from an overexpressing Escherichia coli strain. The enzyme was dimeric, and its optimum pH was 7.0-7.5. Phylogenetic analysis showed the close relationship of this enzyme to other hydrolases involved in the degradation of monocyclic aromatic compounds, and this enzyme was specific for 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid (6-phenyl-HODA), having little activity toward 2-hydroxy-6-oxohepta-2,4-dienoic acid and 2-hydroxymuconic semialdehyde. The enzyme had a Km of 2.51 μM and kcat of 2.14 (s−1) for 6-phenyl-HODA (50 mM sodium phosphate, pH 7.5, 25°C). The effect of the presence of an amino group or hydroxyl group at the 2′-position of phenyl moiety of 6-phenyl-HODA on the enzyme activity was found to be small; the activity decreased only in the order of 6-(2′-aminophenyl)-HODA (2.44 U/mg)>6-phenyl-HODA (1.99 U/mg)>2-hydroxy-6-oxo-6-(2′-hydroxyphenyl)-hexa-2,4-dienoic acid (1.05 U/mg). The effects of 2′-substitution on the activity were in accordance with the predicted reactivity based on the calculated lowest unoccupied molecular orbital energy for these substrates.

Original languageEnglish
Pages (from-to)36-45
Number of pages10
JournalBioscience, Biotechnology and Biochemistry
Volume67
Issue number1
DOIs
Publication statusPublished - 2003 Jan 1

Keywords

  • Carbazole
  • Dibenzofuran
  • Meta-cleavage compound hydrolase
  • Pseudomonas resinovorans strain CA10

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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  • Cite this

    Nojiri, H., Taira, H., Iwata, K., Morii, K., Nam, J. W., Yoshida, T., Habe, H., Nakamura, S., Shimizu, K., Yamane, H., & Omori, T. (2003). Purification and Characterization of meta-Cleavage Compound Hydrolase from a Carbazole Degrader Pseudomonas resinovorans Strain CA10. Bioscience, Biotechnology and Biochemistry, 67(1), 36-45. https://doi.org/10.1271/bbb.67.36