摘要Objective To prepare an active anti-tumor component, compound K (C-K), from saponins in leaves of Panax notoginseng (SLPN) using immobilized β-glucanase. Methods Two entrapments, alginate gel-1 (Alg 1) and alginate gel-2 (Alg 2), were evaluated for their ability to immobilize β-glucanase. The amount and purity of C-K obtained from the transformation process were analyzed by HPLC, and the immobilizing parameters were optimized. Results β-Glucanase can be immobilized and reused with either of the entrapment. However, using AIg 1 resulted in higher enzyme activity than Alg 2. The optimal concentration of the immobilized enzyme was 10%; The optimal crosslinking time was 4-6 h; and the optimal concentration of the crosslinking agent was 6%-7%. Conclusion Immobilized β-glucanase shows sustained enzyme activity, good ethanol tolerance, and was reusable for the preparation of C-K from SLPN.