Aromatization of 4-oxocyclohexanecarboxylic acid to 4-hydroxybenzoic acid by two distinctive desaturases from Corynebacterium cyclohexanicum. Properties of two desaturases.
We have previously demonstrated that Corynebacterium cyclohexanicum degrades cyclohexanecarboxylic acid, a bacteriocide, through a pathway including the aromatization of 4-oxocyclohexanecarboxylic acid to 4-hydroxybenzoic acid [Kaneda, T. (1974) Biochem. Biophys. Res. Commun. 58, 140-144]. Aromatization has now been shown to be catalysed by two desaturase enzymes. Under the action of desaturase I, 4-oxocyclohexanecarboxylic acid is converted to (+)-4-oxocyclohex-2-enecarboxylic acid which is then aromatized by desaturase II to 4-hydroxybenzoic acid. The latter reaction is presumed to occur via the unstable intermediate, 4-oxocyclohex-2,5-dienecarboxylic acid, which is spontaneously isomerized to 4-hydroxybenzoic acid. Desaturase I has been purified in an electrophoretically homogeneous form. It is monomeric with a molecular mass of 67 kDa and contains one tryptophan, one histidine and two cysteine residues per enzyme molecule. The enzyme produces an equivalent amount of 4-oxocyclohex-2-enecarboxylic acid and hydrogen peroxide from 4-oxocyclohexanecarboxylic acid. The properties of desaturase I have been studied in detail. Desaturase II is unstable and has been partially purified. Its characterization is therefore limited. However, the molecular mass of desaturase II was estimated to be 43 kDa by gel filtration chromatography. The characterization of both desaturase enzymes is described in this paper. The possible environmental importance of microbial aromatization in the biodegradation of compounds with the cyclohexane structure is discussed.
Kaneda
T
T
Alberta Research Council, Edmonton, Canada.
Obata
H
H
Tokumoto
M
M
D000596:Amino Acids / Q000032:analysis; D001141:Aromatase / Q000378:metabolism; D003352:Corynebacterium / Q000201:enzymology*; D003470:Culture Media; D003509:Cyclohexanecarboxylic Acids / Q000737:chemistry / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010088:Oxidoreductases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010226:Parabens / Q000378:metabolism*; D013379:Substrate Specificity
D000596:Amino Acids; D003470:Culture Media; D003509:Cyclohexanecarboxylic Acids; D010226:Parabens; C008225:4-ketocyclohexanecarboxylic acid; C085240:4-oxocyclohex-2-enecarboxylate 5-desaturase; D010088:Oxidoreductases; D001141:Aromatase; C085238:4-oxocyclohexanecarboxylate 2-desaturase; C038193:4-hydroxybenzoic acid