Activity and partial purification of chlorophyllase in aqueous systems.

Enzymatic hydrolysis of chlorophylls into chlorophyllides and phytol, and transesterification to the corresponding methyl and ethyl esters, was reported by WillstB;tter and Stoll in 1913 (1). Krossing (2) found such activity localized in the chloroplasts, while more recently Ardao and Vennesland (3) showed that spinach chloroplastin exhibits chlorophyllase activity. In these and other (4) investigations, however, chlorophyllase had been demonstrated only in crude preparations such as acetone powders and pastes and only in reaction mixtures containing a high proportion of organic solvents. Holden (5) recently succeeded in extracting some sugar beet chlorophyllase with citrate and achieved further purification by acetone precipitation. We have partially purified the enzyme from etiolated rye chloroplasts, and determined some of its properties in an aqueous assay system.