Investigation of purification procedures to isolate rat mitochondrial ẟ - aminolaevulinic acid synthetase

Abstract

1. A Sample of rat liver mitochondrial ALV-Synthetase was purified to a specific activity of 4,684 units/ mg, the highest activity yet observed from a mammalian source. 2. The sequence of purification steps that permitted the isolation of the high specific activity enzyme noted above was developed during the work reported here. The sequence of procedures finally used to purify a mitochondrial extract included 0-50% ammonium sulphate precipitation, 5-20% (w/v) polyethylene glycol precipation, CM-sephadex chromatography, Sephadex G-100 filtration, and electrophoresis. 3. Attempts to duplicate previously reported purifications of ALV-Synthetase by use of affinity chromatography were unsuccessful. 4. Isoelectric focusing gave no clearly useful or preparative separations of ALV-Synthetase in pH gradients.