Artículo
Identification of an extradiol dioxygenase involved in tetralin biodegradation: Gene sequence analysis and purification and characterization of the gene product
Autor/es | Andújar, Eloísa
Hernáez, María José Kaschabek, Stefan R. Reineke, Walter Santero Santurino, Eduardo |
Departamento | Universidad de Sevilla. Departamento de Genética |
Fecha de publicación | 2000-02 |
Fecha de depósito | 2017-07-24 |
Publicado en |
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Resumen | A genomic region involved in tetralin biodegradation was recently identified in Sphingomonas strain TFA. We have cloned and sequenced from this region a gene designated thnC, which codes for an extradiol dioxygenase required ... A genomic region involved in tetralin biodegradation was recently identified in Sphingomonas strain TFA. We have cloned and sequenced from this region a gene designated thnC, which codes for an extradiol dioxygenase required for tetralin utilization. Comparison to similar sequences allowed us to define a subfamily of 1,2-dihydroxynaphthalene extradiol dioxygenases, which comprises two clearly different groups, and to show that ThnC clusters within group 2 of this subfamily. 1,2-Dihydroxy-5,6,7,8-tetrahydronaphthalene was found to be the metabolite accumulated by a thnC insertion mutant. The ring cleavage product of this metabolite exhibited behavior typical of a hydroxymuconic semialdehyde toward pH-dependent changes and derivatization with ammonium to give a quinoline derivative. The gene product has been purified, and its biochemical properties have been studied. The enzyme is a decamer which requires Fe(II) for activity and shows high activity toward its substrate (V(max), 40.5 U mg-1 K(m) 18.6 μM). The enzyme shows even higher activity with 1,2-dihydroxynaphthalene and also significant activity toward 1,2-dihydroxybiphenyl or methylated catechols. The broad substrate specificity of ThnC is consistent with that exhibited by other extradiol dioxygenases of the same group within the subfamily of 1,2- dihydroxynaphthalene dioxygenases. |
Agencias financiadoras | Comisión Interministerial de Ciencia y Tecnología (CICYT). España European Union (UE) |
Identificador del proyecto | BIO96-0908
EV5V-CT92-0192 |
Cita | Andújar, E., Hernáez, M.J., Kaschabek, S.R., Reineke, W. y Santero, E. (200-). Identification of an extradiol dioxygenase involved in tetralin biodegradation: Gene sequence analysis and purification and characterization of the gene product. Journal of Bacteriology, 182 (3), 789-795. |
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