Immobilized ACP and electrofocusing used to purify the palmityl-CoA-ACP-transacylase from Mycobacterium smegmatis.

This paper describes the first purification to electrophoretic and chromatographic homogeneity of a palmityl-CoA-ACP-transacylase. Palmityl-CoA-ACP-transacylase of Mycobacterium smegmatis shows two interconvertible forms in monomer-oligomer correlation. Taking advantage of this interconvertibility properties a 800-fold purification of the enzyme was recently achieved (Kervabon et al (1976) Biochimie, 58, 647-656). The present report describes the use of immobilized ACP and electrofocusing to obtain a 880-fold and a 5 500-fold purification respectively. The enzyme thus purified, pI 4.7, showed a single band upon polyacrylamide gel electrophoresis. Chromatography on Sephadex G-150 revealed only the peak of the monomer. When the enzyme solution containing 200 microng/ml of protein was concentrated by vacuum dialysis to approximatively 1 mg/ml, gel electrophoresis showed two bands one corresponding to the monomer and the other to the oligomer formed.