β-葡萄糖苷酶固定化设计与合成稀有人参皂苷的研究.pdf

Abstract Study of β- glucosidase immobilized design and synthesis of rare ginsenosides β-glucosidase (Tnap0602) derived from Thermotoga Naphthophila RUK-10 can efficiently transform ginsenoside Rb1 to form rare ginsenoside Rd. In this paper, a novel β-glucosidase (Carbamate-binding module) The method of immobilization was used to study the immobilization method of heteroenzyme, which laid the foundation for the application of β-glucosidase. In this paper, two engineered bacteria capable of expressing the fusion proteins 0602-CBM2 and 0602-CBM3 were constructed and the target protein was purified. The purified protein was adsorbed to microcrystalline cellulose, regenerated amorphous cellulose and filter paper pulp, respectively. The preliminary immobilization of β-glucosidase was achieved and the ginsenoside conversion experiment was carried out. The results showed that the maximum binding capacity of microcrystalline cellulose of recombinant enzyme 0602-CBM2 was 2mg / ml, while 0602-CBM3 did not bind to microcrystalline cellulose, regenerated amorphous cellulose and filter paper pulp. The results showed that the immobilized β-glucosidase could be used in the re-use of immobilized enzyme, and the activity of immobilized β-glucosidase could be improved after the repeated use of immobilized enzyme 0602-CBM2 for three times , The HPLC assay showed that a single product peak was evident at 35 minutes compared to the extract without the enzyme. Keywords: β-glucosidase, cellulose-binding modules, immobilization, ginsenoside II