Methods for the Extraction of Enzymes from Cereal Leaves with Especial Reference to the Triosephosphate Dehydrogenases.

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Abstract
Methods were developed for the extraction and purification of enzymes, especially triosephosphate dehydrogenases, from immature and mature cereal leaves. Two facets of this work were: 1st, the addition of cysteine to the grinding medium for the preparation of active acetone powders or aqueous extracts; 2d the use of Carbowax 4000 as 1 of the enzyme precipitants. Acetone fractionation was used for the final stages of purification. These techniques permit the preparation of highly purified triosephosphate dehydrogenases in good yields from small (100 g) of leaf tissue. Qualitative tests demonstrated that these methods were suitable for the extraction of many other enzymes from cereal leaves. The preparative steps are simple and rapid and are recommended for routine investigation of leaf enzymes, as well as for methods that are suitable for the classroom demonstration of the principles of enzyme extraction, purification and assay.