Synthesis of [formula omitted] coenzyme a with a partially purified [formula omitted]:CoA ligase from cell suspension cultures of soybean ( Glycine max)

Abstract

1. 1. A p- coumarate : CoA ligase has been isolated from cell suspension cultures of soybean. Partial purification of the enzyme achieved by MnCl 2 and (NH 4) 2SO 2 precipitation and by gel chromatography on Sephades G-100. 2. 2. Under the conditions of the hydroxamic acid assay, the enzyme has a pH optimum at about pH 8.5 and a temperature optimum at about 30°C. It requires Mg 2+ and ATP at a molar ratio of 1.1. The molecular weight of the enzyme was estimated from the elution volume after chromatography on a Sephadex G-100 column to be about 55 000. 3. 3. Of the various cinnamic acids tested, p- coumarate was the most effective substrate. 4. 4. The partially purified p- coumarate :CoA ligase was utilized for the enzymatic synthesis of p- coumaroyl CoA. Gel chromatography on Sephadex G-10 proved to be a simple and rapid procedure for the isolation and purification of the product.