Purification and Properties of Chlorophyllase from Ailanthus altissima (Tree-of-Heaven)

Plant Physiol. 1971 May; 47(5): 609–618.

PMCID: PMC396737

Purification and Properties of Chlorophyllase from Ailanthus altissima (Tree-of-Heaven) ^1,²

Roger F. McFeeters,³ Clinton O. Chichester, and John R. Whitaker

^aDepartment of Food Science and Technology, University of California, Davis, California 95616

³ Present address: Department of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan 48823.

¹ This investigation was supported in part by the National Institutes of Health (TO1U1-01053 and AM-11665).

² Taken from the dissertation submitted by Roger F. McFeeters to the Graduate Division, University of California, Davis, in partial satisfaction of the requirements for the Ph.D. degree.

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Abstract

Chlorophyllase from Ailanthus altissima leaves has been purified 63-fold by a combination of heat treatment, ultracentrifugation, gel filtration, and chromatography on diethylaminoethyl cellulose. While the enzyme is inhibited to some degree by Triton X-100, a modification of the assay procedure of Klein and Vishniac has been shown to be far superior to the use of aqueous acetone systems. The enzyme was found to have a pH optimum on pheophytin a of 4.5. Chlorophylls a and b, pheophytins a and b, and pyropheophytin a were hydrolyzed by the enzyme while protochlorophyll a and 4-vinyl protochlorophyll a were not hydrolyzed but were competitive inhibitors. p-Nitrophenyl acetate was not hydrolyzed. The enzyme does not appear to contain an essential sulfhydryl group since sodium tetrathionate and p-chloromercuribenzoate did not affect its activity.

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Selected References

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