USE OF VESICULAR CHROMATOGRAPHY TO PURIFY PLANT ENZYMES

A method for fast purification of rubisco and isocitrate lyase (ICL) t o a homogeneous state based on use of vesicular chromatography is desc ribed in the present paper. Purification of rubisco involved precipita tion of the enzyme by PEG 4000 at 15% saturation with subsequent separ ation on a column filled with the chromatographic material Vesipor. Th e boundary of Vesipor permeability was raised by means of preliminary treatment with 2% Na2CO3 for a period of 5 h at 50-degrees-C. Precipit ation by ammonium sulfate (35-50%) can be substituted for precipitatio n with the aid of PEG. Rubisco from leaves of wheat (Triticum vulgare L.), oats (Avena sativa L.), and cucumber (Cucumis sativus L.) and fro m plants of Wolffia arrhiza (L.) Horkel was purified. Purification of ICL from the scutellum of corn (Zea mays L.) involved precipitation by ammonium sulfate (0-30%), vesicular chromatography on a column contai ning Vesipor pretreated with 2% Na2CO3 for 3 h at 20-degrees, and adso rption of protein impurities on DEAE-cellulose. Both enzymes were elec trophoretically homogeneous. Purification was fast (1-2 h) with a high er yield than in former methods.