Purification and properties of 2'-hydroxybenzalpyruvate aldolase from a bacterium that degrades naphthalenesulfonates

• Published in: The Journal of biological chemistry Vol. 268; no. 13; pp. 9484 - 9489
• Main Authors: KUHM, A. E, KNACKMUSS, H.-J, STOLZ, A
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 05.05.1993
• Subjects: Aldehyde-Lyases - antagonists & inhibitors; Aldehyde-Lyases - isolation & purification; Aldehyde-Lyases - metabolism; Aldehydes - pharmacology; Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Biotransformation; Chromatography, Gel; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Kinetics; Miscellaneous; Molecular Sequence Data; Naphthalenes - metabolism; Naphthalenesulfonates - metabolism; Pseudomonas; Pseudomonas - enzymology; Spectrophotometry; Substrate Specificity; Pseudomonadales; Purification; Enzymatic activity; Enzyme; Reaction mechanism; Bacteria; Pseudomonadaceae; Pseudomonas; Naphthalene derivatives; Sulfonate
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(18)98376-6

2'-Hydroxybenzalpyruvate aldolase catalyzes the cleavage of 2'-hydroxybenzalpyruvate to salicylaldehyde and pyruvate. This reaction is part of the degradative pathways for naphthalene and naphthalenesulfonates by bacteria. 2'-Hydroxybenzalpyruvate aldolase has been purified to homogeneity from a bacterium that degrades naphthalenesulfonates (strain BN6). The enzyme has a molecular weight of about 120,000 and is composed of identical subunits with a molecular weight of about 38,500. Thus the enzyme appears to exist as a trimeric oligomer. The NH2-terminal amino acid sequence did not show significant homology to other published amino acid sequences. Extensive loss of enzyme activity occurred when the enzyme was incubated with 2'-hydroxybenzalpyruvate in the presence of sodium borhydride. This suggested the intermediate formation of a stable Schiff base between enzyme and substrate. 2'-Hydroxybenzalpyruvate aldolase was inhibited by p-chloromercuribenzoate and by the reaction product salicylaldehyde. The enzyme converted 2'-hydroxybenzalpyruvate, 2',4'- and 2',6'-dihydroxybenzalpyruvate.