Multiple forms of endo-1,4-b-glucanases in the endosperm of Euphorbia heterophylla L.

Germinating seeds of Euphorbia heterophylla L. contain endo-1,4-b-glucanases which degrade carboxymethylcellulose (CMC). The activity decreased approximately 66 % in extracts of endosperm containing isopropanol or ethanol. The endoglucanases were isolated from endosperm extracts using ammonium sulphate fractionation followed by Sephacryl S-100-HR chromatography resulting in two main peaks: I and II. Peak I endoglucanase was further puri®ed about 15-fold on DEAE-Sephadex A50 and then by af®nity chromatography (CF11-cellulose). Peak II endoglucanases were further puri®ed 10-fold on CMcellulose chromatography. The results indicated the occurrence of a 66 kDa endoglucanase (fractionated by SDS-PAGE and visualized by activity staining using Congo Red). Several acidic (pI 3.0 to 5.7) and basic (pI 8.5 to 10.0) forms from both peaks which differed in their capacities for degrading CMC or xyloglucans from Copaifera langsdorf®i or Hymenaea courbaril were detected