Abstract: Two kinds of β-Glucosidases from Camellia sinensis var. sinensis cv. Yabukita were purified by a sophisticate application of affinity chromatography. An affinity adsorbent for β-Glucosidases has been prepared with a positively charged β-glucosylamidine as a ligand in which a glycon moiety is connected via a N-glycoside linkage. The synthesized affinity adsorbent is quite stable under mildly acidic conditions (pH5~6) and can be used up to 20 times over three months without significant affect on the column performance. Two tea leaf β-Glucosidases have been purified to be monomers with molecular mass of 63 and 75 kDa by affinity chromatography at pH6.0 and pH5.0, respectively. The extensive characterization of tea leaf β-Glucosidasse has been reported. Tea aroma precursors such as benzyl, (Z)-3-hexenyl and D-mandelonitrile β-D-glucopyranosides were hydrolyzed by the purified two β-Glucosidases to liberate tea aroma.

Key words: Tea science, β-Glucosidase, Tea aroma, Affinity chromatography, Camellia sinensis var, Sinensis cv, Yabukita, β-Glucosylamidine