Abstract: The α-L-rhamnosidase plays an important role in the conversion of natural products, and Pichia pastoris was used to express α-L-rhamnosidase AnRhaE. The pPIC9K-AnRhaE expression vector was constructed and transformed into Pichia pastoris GS115 and KM71H. After induced expression, SDS-PAGE electrophoresis showed that GS115 was more conducive to the production of AnRhaE. High AnRhaE activity was obtained at 45-55 ℃, and the optimum temperature was 55 ℃. AnRhaE was stable at pH 4.5-8, and the optimum pH was 5. AnRhaE could hydrolyze α-1,2, α-1,6 and rhamnosyl-rhamnosyl glycosidic bond, including naringin dihydrogen chalcone to trilobatin, naringin to prunin, rutin to isoquercetin, neohesperidin to hesperetin-7-O-glucoside, hesperidin to hesperetin-7-O-glucoside and epimedin C to icariin. AnRhaE had high activity to α-1,2 glycosidic bond than the α-1,6 glycosidic bond, and could complete hydrolysis epimedin C to icariin. Therefore, AnRhaE is potentially applicable in enzymatic conversion of natural products.
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