2-AMINOBENZOYL-COA MONOOXYGENASE REDUCTASE, A NOVEL TYPE OF FLAVOENZYME - IDENTIFICATION OF THE REACTION-PRODUCTS

In a previous report we have described some properties of a novel flavoenzyme from a denitrifying Pseudomonas species which catalyzes the oxygen‐ and NAD(P)H‐dependent conversion of 2‐aminobenzoyl‐CoA [Buder, R., Ziegler, K., Fuchs, G., Langkau, B. & Ghisla, S. (1989) Eur. J. Biochem. 185, 637–634]. In this paper, we report on the identification of the three products formed from 2‐aminobenzoyl‐CoA in this reaction. The spectroscopic data and the chemical properties of these compounds and those of their degradation products are compatible with the structures of 2‐amino‐5‐hydroxybenzoyl‐CoA, 2‐amino‐5‐hydroxycyclohex‐1‐enecarboxyl‐CoA and of 2‐amino‐5‐oxocyclohex‐1‐enecarboxyl‐CoA. The latter is the main product and was found to be rather unstable since it hydrolyzes and decarboxylates readily at pH < 5. Ammonia is released from the decarboxylation product in the neutral pH range to yield 1,4‐cyclohexanedione. Conditions were optimized such that the CoA thioester of 2‐amino‐5‐hydroxybenzoate is the product obtained at > 98% yield. 2‐amino‐5‐hydroxycyclohex‐1‐enecarboxyl‐CoA is the product which is formed when the mixture of the reaction products is treated with sodium borohydride before separation. Copyright © 1990, Wiley Blackwell. All rights reserved