Open AccessPurification and biochemical characterization of gentisate 1,2‐dioxygenase from Klebsiella pneumoniae M5a1
Author(s) -
Suárez, Mónica,
Ferrer Estrella,
Martín Margarita
Publication year - 1996
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.1574-6968.1996.tb08466.x
Subject(s) - klebsiella pneumoniae , microbiology and biotechnology , enterobacteriaceae , klebsiella , dioxygenase , chemistry , biology , escherichia coli , biochemistry , enzyme , gene
Gentisate 1,2‐dioxygenase (E.C.I.14.13) was purified to homogeneity from Klebsiella pneumoniae M5a1, a soil bacterium able to degrade a great variety of aromatic compounds. The molecular mass of the purified holoenzyme was 159 kDa and its structure was deduced to be a tetramer with 38 kDa per subunit. Gentisate 1,2‐dioxygenase appears to contain Fe 2+ in its active site. The optimum temperature for enzyme activity was estimated to be 30 °C, the optimum pH values varied between 8 and 9 and the isoelectric point was 4.7. Gentisate dioxygenase exhibited typical saturation kinetics and had an apparent K m of 52 μM for gentisate. Its amino acid content was determined to be very similar to that of the enzyme from Pseudomonas acidovorans .