Producing Ginsenoside Compound-K By The Microorganism Transformation

The purpose of this paper is to study the characterization of the ginsenosidase which can hydrolyze ginsenoside the protopanaxadiol type saponin (PPD) to ginsenoside compound K(C-K), the ginsenoside compound K(C-K) from enzyme reaction was isolated by silica gel column chromatography, the purity was examined by the method of HPLC.In order to get the strain producting the enzyme which could hydrolyse the PPD to the ginsenoside compound K(C-K) as much as possible, we selected4strains to study the ability of producting ginsenside-glycosidase, sp.3, sp.18, sp.48, sp.848. The experiment found sp.848yielded the highest ginsenside-glycosidase, Futhermore, it was shown that7days was the best period of enzyme fermentation.The enzyme isolated from sp.848was purified by a column of DEAE-Cellulose and polyacrylamide gel electrophoresis, and the characteration of enzyme were studied. After purified by a column of DEAE-Cellulose and molecular weight with the method of SDS-polyacrylamide gel electrophoresis, the molecular weight was74kDa different from the molecular weight77kDa studied by Wang Liang. The results showed the optimum pH was5.0, and the optimum temperature was at45℃. The enzyme which is ginsenoside glycosidase I can hydrolyse the ginsenoside Rb1, Rb2, Rc to the ginsenoside compound K(C-K) well, but can not hydrolyse the ginsenoside Rd well.Refering to the fittest condition we enlarged the enzyme reaction. The decolorizing by D-296removed4.74%impurity, and ungreasing esters by petroleum benzin was removed7.46%impurity. Finally, it was purified by the Silica gel column. The best mobile phase was the mixture of V(chloroform):V(methanol)=9.5:0.5. We get the ginsenoside C-K138g from crude ginsenoside C-K335g. In the silica gel column, ginsenoside C-K was obtained41.2%. After crystallizating, the pure C-K yield was90%. The ginsenoside C-K purity was97%by the method of HPLC.