Expression, purification, and characterization of 2′-aminobiphenyl-2, 3-diol 1,2-dioxygenase from carbazole-degrader Pseudomonas resinovorans strain CA10

Kenichi Iwata, Hideaki Nojiri, Kentaro Shimizu, Takako Yoshida, Hiroshi Habe, Toshio Omori

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The two-subunit meta-cleavage enzyme, 2′-aminobiphenyl-2,3-diol 1,2-dioxygenase (CarBaBb), from the carbazole degrader Pseudomonas resinovorans strain CA10 was purified to homogeneity from an Escherichia coli strain carrying the expression vector pUCA503, in which two copies of the carBaBb genes are tandemly linked. SDS-PAGE and gel filtration showed that CarB was a α2β2-heterotetrameric enzyme with subunit molecular masses of approximately 10,000 for CarBa and 29,000 for CarBb. The optimum pH for activity was 8.5 and that of temperature was 35°C. The CarB enzyme had a Km of 14 μM and a kcat/Km of 0.25 μM-1 s-1 for 2′-aminobiphenyl-2,3-diol, and the catalytic activities for biphenyl-type catecholic substrates were higher than those for monoaromatic catechol derivatives. The enzyme was originally isolated as a meta-cleavage enzyme for 2′-aminobiphenyl-2,3-diol involved in carbazole degradation, but the enzyme was highly specific for 2,3-dihydroxybiphenyl.

Original languageEnglish
Pages (from-to)300-307
Number of pages8
JournalBioscience, Biotechnology and Biochemistry
Volume67
Issue number2
Publication statusPublished - 2003 Feb
Externally publishedYes

Fingerprint

Pseudomonas resinovorans
carbazoles
Dioxygenases
glycols
Purification
Enzymes
enzymes
catechol
Molecular mass
catalytic activity
biphenyl
Escherichia coli
2-aminodiphenyl
carbazole
Catalyst activity
Gels
Genes
chemical derivatives
gels
molecular weight

Keywords

  • Biodegradation
  • Carbazole
  • Meta-cleavage enzyme
  • Pseudomonas resinovorans
  • Purification

ASJC Scopus subject areas

  • Food Science
  • Applied Microbiology and Biotechnology
  • Chemistry (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Bioengineering

Cite this

Expression, purification, and characterization of 2′-aminobiphenyl-2, 3-diol 1,2-dioxygenase from carbazole-degrader Pseudomonas resinovorans strain CA10. / Iwata, Kenichi; Nojiri, Hideaki; Shimizu, Kentaro; Yoshida, Takako; Habe, Hiroshi; Omori, Toshio.

In: Bioscience, Biotechnology and Biochemistry, Vol. 67, No. 2, 02.2003, p. 300-307.

Research output: Contribution to journalArticle

Iwata, Kenichi ; Nojiri, Hideaki ; Shimizu, Kentaro ; Yoshida, Takako ; Habe, Hiroshi ; Omori, Toshio. / Expression, purification, and characterization of 2′-aminobiphenyl-2, 3-diol 1,2-dioxygenase from carbazole-degrader Pseudomonas resinovorans strain CA10. In: Bioscience, Biotechnology and Biochemistry. 2003 ; Vol. 67, No. 2. pp. 300-307.
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