Enzyme catalysed non-oxidative decarboxylation of aromatic acids. II. Identification of active site residues of 2,3-dihydroxybenzoic acid decarboxylase from Aspergillus niger.
In order to understand the mechanism of decarboxylation by 2,3-dihydroxybenzoic acid decarboxylase, chemical modification studies were carried out. Specific modification of the amino acid residues with diethylpyrocarbonate, N-bromosuccinimide and N-ethylmaleiimide revealed that at least one residue each of histidine, tryptophan and cysteine were essential for the activity. Various substrate analogs which were potential inhibitors significantly protected the enzyme against inactivation. The modification of residues at low concentration of the reagents and the protection experiments suggested that these amino acid residues might be present at the active site. Studies also suggested that the carboxyl and ortho-hydroxyl groups of the substrate are essential for interaction with the enzyme.
Kamath
A V
AV
Department of Biochemistry, Indian Institute of Science, Bangalore.
Rao
N A
NA
Vaidyanathan
C S
CS
D001234:Aspergillus niger / Q000201:enzymology*; D001665:Binding Sites; D001974:Bromosuccinimide / Q000494:pharmacology; D002262:Carboxy-Lyases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D007700:Kinetics
D002262:Carboxy-Lyases; C022501:2-pyrocatechuate decarboxylase; D001974:Bromosuccinimide; D004047:Diethyl Pyrocarbonate; D005033:Ethylmaleimide