Abstract
SUMMARY A procedure is described for the 500-fold purification of chloro- phyllase from etiolated rye seedlings. Evidence is presented that the final preparation is a lipoprotein-sodium deoxycholate complex. An assay procedure with an aqueous medium is presented. p-Chloromercuribenzoate and ferricyanide inhibit the enzyme, whereas iodoacetamide and diisopropyl fluorophos- phate do not, at the concentrations tested. The possible role of the enzyme in the biosynthesis and breakdown of chlorophyll and in the formation of lamellas of the chloroplast is discussed. REFERENCES 1. 2. 3. k: 6. 7. 8. 9. 10. 11. 12. 13. 14. WILLST~TTER, R., AND STOLL, A., Untersuchungen Ober Chloro- phyll, Springer-Verlag, Berlin, 1913, p. 172. KROSSING, G., Biochem. Z., 306, 359 (1940). ARDAO, C., AND VENNESLAND, B., Plant Physiol., 36, 368 (1960). MAYER, H., Planta, 11, 294 (1930). HOLDEN, M., Biochem. J., 78, 359 (1961). WEAST, C.
Highlights
Source of Enzyme-In agreement with previous reports we found chlorophyllase activity in Ailanthus, parsley, stinging nettle, and dandelion leaves
Barley and oat seedlings grown in the laboratory were very poor sources, but etiolated rye seedlings (Detra-Petkus variety of winter rye’) grown for 7 to 8 days proved superior in specific activity to other plants tested and were adopted as starting material. -4 preparation with good activity was obtained from Euglena grads
The very small amount of chlorophyll present in chloroplasts grown in the dark was completely removed in the course of enzyme purification
Summary
Source of Enzyme-In agreement with previous reports we found chlorophyllase activity in Ailanthus, parsley, stinging nettle, and dandelion leaves. In the course of screening various sources for chlorophyllase, the method of Weast and Mackinney (6) was employed In this method, a weighed amount (10 to 20 mg) of leaf acetone powder is freed of chlorophyll and incubated for various periods (3 to 18 hours) in 66% aqueous acetone containing a known amount of substrate. The assay procedure devised by us is based on that of Weast and Mackinney, but with the significant difference that the reaction takes place in an entirely aqueous medium. This t Postdoctoral Fellow, National Cancer Institute, United States Public Health Service. Assay conditions were the same as in Table I. step Preparation
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