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 language | English |
|---|---|
| Pages (from-to) | 36-45 |
| Number of pages | 10 |
| Journal | Bioscience, Biotechnology and Biochemistry |
| Volume | 67 |
| Issue number | 1 |
| Publication status | Published - 2003 Jan |
| Externally published | Yes |
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Keywords
- Carbazole
- Dibenzofuran
- Meta-cleavage compound hydrolase
- Pseudomonas resinovorans strain CA10
ASJC Scopus subject areas
- Food Science
- Applied Microbiology and Biotechnology
- Chemistry (miscellaneous)
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biotechnology
- Bioengineering
Cite this
Purification and characterization of meta-cleavage compound hydrolase from a carbazole degrader Pseudomonas resinovorans Strain CA10. / Nojiri, Hideaki; Taira, Hiroko; Iwata, Kenichi; Morii, Kenichi; Nam, Jeong Won; Yoshida, Takako; Habe, Hiroshi; Nakamura, Shugo; Shimizu, Kentaro; Yamane, Hisakazu; Omori, Toshio.
In: Bioscience, Biotechnology and Biochemistry, Vol. 67, No. 1, 01.2003, p. 36-45.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Purification and characterization of meta-cleavage compound hydrolase from a carbazole degrader Pseudomonas resinovorans Strain CA10
AU - Nojiri, Hideaki
AU - Taira, Hiroko
AU - Iwata, Kenichi
AU - Morii, Kenichi
AU - Nam, Jeong Won
AU - Yoshida, Takako
AU - Habe, Hiroshi
AU - Nakamura, Shugo
AU - Shimizu, Kentaro
AU - Yamane, Hisakazu
AU - Omori, Toshio
PY - 2003/1
Y1 - 2003/1
N2 - 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.
AB - 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.
KW - Carbazole
KW - Dibenzofuran
KW - Meta-cleavage compound hydrolase
KW - Pseudomonas resinovorans strain CA10
UR - http://www.scopus.com/inward/record.url?scp=0012484786&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0012484786&partnerID=8YFLogxK
M3 - Article
C2 - 12619671
AN - SCOPUS:0012484786
VL - 67
SP - 36
EP - 45
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
SN - 0916-8451
IS - 1
ER -