Abstract: The aim of this study was to separate and purify alliinase from fresh garlic,and determine the enzymatic properties of alliinase. The optimal separation and purification process is as follows:to extract alliinase from fresh garli with Na+/K+ phosphate buffer(pH 7.0),salt out it with 45% saturated ammonium sulfate,separate it by centrifuge with 10000×g,dissolve precipitate in Na+/K+ phosphate buffer(pH 6.5) and dialyze using dialysis bag with 14000 molecular weight cutoff,purify alliinase with Sephadex G-200 column,and collect eluate when elution time is up to 4.75 h. The optimum temperature of alliinase is 30℃,and the optimum pH is 6.3. Mg2+,Zn2+,Fe2+,Fe3+,and EDTA-Na have active effects on alliinase,and the effect of Fe3+ is specially obvious,but alliinase would be seriously inhibited by Cu2+. The Km and Vmax values of alliinase are 5.91 mmol/L and 1.55μmol/min respectively when synthetical S-allyl-L-cysteine sulphoxide is used as basic substrate.

Key words: alliinase, purification, enzymatic properties