Crude extract of sunflower seeds was prepared using different extractants (distilled water, 5% NaCl in distilled water, 5% NaCl in acetate buffer pH 5.0) and certain mixing ratios (1:2, 1:5 and 1:10 seeds powder : extractant, respectively). Using of 5% NaCl in acetate buffer pH 5.0 at mixing ratio of 1:5 gave an extract with reasonable efficient clotting properties. Ammonium sulphate at 40-60% concentration was used in one single step for the partial purification of the extract. Ammonium sulphate, (40-60%) fraction showed milk clotting and specific activities of 440 sec and 2.30 U/mg protein, respectively. Meanwhile a yield percentage and a fold of purification of 28.58 and 2.02 were achieved. Clotting/proteolytic activity, water holding capacity and curd synersis of the partially purified extract approved those of commercial calf rennet indicating the suitability of the extract as a rennet substitute. The optimum temperature, pH, Cacl 2 and GDL concentration of the resultant extracts were found to be 65°C, 5.0, 0.04 % and 0.4%, respectively.
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