Special Ginsenosidases From Different Microorganism Strains

Ginsenoside Rg3is one of important physiological activity ingredients in ginseng, plays an important role in anti-tumor, belongs to protopanaxadiol type saponin. In this article, transforming protopanaxadiol type saponin to ginsenoside Rg3by ginsenosidase from different microorganism strains was studied.Firstly,7types of ginsenosides were extracted from three kinds of ginseng analyzing by Spectrophotometer and HPLC, and the ginsenosides Rb1, Rb2, Rc, Rd, Re Rf and Rgi were the main composition. The total yield of the7types of ginsenosides was up to90%. Red ginseng contained more saponins than the white ginseng; the kinds of the saponin in the top of the root were more than the main root.To obtain the proper ginsenosidase which could hydrolyze protopanaxadiol type saponins into ginsenoside Rg3, one fungus and6kinds of bacterium from KAIST were selected. TLC was used to detect the conditions of fermentation, such as types of inducer in medium, concentration of inducer, the optimal fermented time, the optimal fermented temperature et al. The results were shown that the optimal fermented conditions for the new bacterium Phycicoccus ginsenosidimutans BXN5-13T are temperature(30℃),time (48h),in No.III culture medium with0.02%ginsenoside as inducer. The bacterium with the gene of Esherichia coli.was chosed.DEAE-cellulose column was used to purify the crude enzyme from sp.GS3082Rhodanobacter ginsenosidimutans to obtain the pure ginsenosidase which could hydrolyze protopanaxadiol type saponins into Rg3. SDS-polyacrylamide gel electrophoresis was used to estimate the molecule weight of ginsenosidase, the result was about70.1kDa. The properties of ginsenosidase were detected also.Lastly,50g the enzymatic reaction product containing ginsenoside Rg3was separated by the AB-8Diaion resin to remove the inorganic ion and sugar, further purified by the Silica Gel to obtain the high purity ginsenoside Rg3. And the result showed that the yield of pure ginsenoside Rg3was8.1542g, the mixture of ginsenoside Rg3and Rd was13.18g. The total yield of ginsenoside Rg3was87.89%.