Structure of the prosthetic group of Klebsiella aerogenes citrate (pro-3S)-lyase.

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Abstract
The prosthetic group of citrate (pro-3S)-lyase [citrate oxaloacetate-lyase (pro-3S-CH2COO- .fwdarw. acetate). EC 4.1.3.6] from K. aerogenes was obtained by mild alkaline hydrolysis of the enzyme and purified by DEAE-cellulose chromatography. Several chemical and enzymatic degradation products of the compound were isolated, and the structure of the prosthetic group was 3''(or 2'') .fwdarw. 1"-(5"-phosphoribosyl) dephospho-Co A. The exact linkage between the 2 ribose moieties and the anomeric configuration of the glycosidic bonds was not determined. A similar analysis was obtained for a product isolated after Pronase digestion of the enzyme without alkaline hydrolysis. The isolated prosthetic group serves as a substrate for a crude preparation of acetate:-SH-(acyl carrier protein) enzyme ligase (AMP) from K. aerogenes, pure pig heart citrate (si)-synthase (EC 4.1.3.7), and pure rat liver ATP citrate (pro-3S)-lyase (EC 4.1.3.8). This compound is the first shown to substitute for coenzyme A in the latter reaction.