β-葡萄糖苷酶的制备以及对人参皂苷的酶法转化-药理学专业论文.docxVIP

III III Abstract Objection ： This experiment We use β -glycosidase fermentatied and purified from Aspergillus to catalytic ginseng saponins and N - acetyl glucosamine to make its formed new glycosidase bond. so as to achieve risk-benefit ratio of new ginseng saponins. These have provide new direction and thinking for saponins biotransformations. β-glycosidase was prepared by the solid-state fermentation，separ- -ation and purification from Aspergillus. The new amino-saponins were formed through the catalysis of ginseng saponins and N - acetyl glucosamine by the β-glycosidase so as to achieve highly effective and low toxicity new ginseng saponins. This study will provide new direction and thinking way for saponins biotransformations. Method: Firstly, the crude enzyme come from Aspergillus purified by Sephadex - G150 and DEAE - Sephadex - and A50 respectivefermentaly to get pureβ-glycosidase. Firstly, the pureβ-glycosidase was obtained from the crude enzyme separated after salting-out, Sephadex - G150 and DEAE - Sephadex - A50. Secondly, After the fresh ginseng was sliced, washed, dried in 50 ℃,then shattered, the ginseng was extracted by using 70% alcoho,ethyl acetate and n-buoh,respectively. The total ginseng saponins were obtained after freeze-drying by the n-butanol extraction after the separation with D101 macroporous resin. Thirdly,the ginseng saponins, N - acetyl glucosamine and β-glycos- -idase were mixed according to the following three method :No.1 routine, ginseng saponins, N - acetyl glucosamine andβ-glycosidase were added simultaneously: No.2 routine , after ginseng saponins and β–glycosidase were mixed for a certain time , N - acetyl glucosamine were add into mixture; No.3 routine, after N - acetyl glucosamine and β–glycosidase were mixed for a certain time , the ginseng saponins were add into reactor. Forthly, total transformed products were purified by (ODS)-C18 reversed-phase silica gel to obtain some parts of transformed products.Using all parts of tr