Open AccessPurification and characterization of the 3‐hydroxybenzoate‐6‐hydroxylase from Klebsiella pneumoniae
Author(s) -
Suárez Mónica,
Ferrer Estrella,
GarridoPertierra Amando,
Martín Margarita
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07431.x
Subject(s) - klebsiella pneumoniae , hydroxybenzoate , microbiology and biotechnology , chemistry , biology , biochemistry , escherichia coli , gene
We isolated 3‐hydroxybenzoate‐6‐hydroxylase (E.C. 1.14.13.), an inducible enzyme that catalyzed the para ‐hydroxylation of 3‐hydroxybenzoate (3‐HBA) to 2,5‐dihydroxybenzoate, from Klebsiella pneumoniae . Although the enzyme was found to be mainly induced by its substrate, a coordinated induction of 3‐hydroxybenzoate hydroxylase and gentisate dioxygenase was also observed in the presence of the product of the reaction. The purified enzyme was a monomer with a molecular mass of 42 000. It contained FAD as a prosthetic group, utilized NADH or NADPH with similar efficiencies and its activity was inhibited by Cu 2+ , Fe 2+ and Hg 2+ . Other properties, such as induction mechanism and kinetic parameters were also studied. Moreover, for the first time the amino acid composition of a 3‐hydroxybenzoate‐6‐hydroxylase was determined.