Ginsenoside Compound-K Production

The purpose of this paper is to study the characterization of the ginsenosidase whichcan hydrolyze ginsenoside the protopanaxadiol type saponin (PPD) to ginsenosidecompound K(C-K), and to isolate and purify ginsenoside C-K. In this paper, two differentenzymes are got from bacterium and fungi separately, and then the protopanaxadiol typesaponin (PPD) is reacted as the substrate with the two different enzymes to produceginsenoside C-K. The purity of the ginsenoside C-K was examined by the method of HPLC.The enzyme isolated from Microbacterium sp.GS514hydrolyzes ginsenoside PPD toRh2, the intermediate production is ginsenoside Rg3. However, the enzyme isolated fromarbiab sp.48hydrolyzes ginsenoside PPD to C-K, the intermediate production isginsenoside F2.In this paper, the characterization of the two different enzymes, the optimumconcentration of substrate, the choice of different medium, the optimum culture time, theoptimum culture pH, and the optimum culture temperature were studied.In the study of ginsenosidase from fungi, the different strain, the different medium, thedifferent inducer were compared by studying the ability of producing ginsenosidase in orderto hydrolyze the PPD as much as possible. The results show that the optimum inducer issophora flower and the optimum fermentation condition is at30℃the strain sp.48fermentby7-8days.In order to get more ginsenoside C-K, the condition of the enzyme reaction was alsostudied. The result showed that the optimum medium was wheat bran: sophora flower=3:1,the optimum concentration of substrate was1%, the optimum culture time of ginsenosidasewas22hr, the optimum temperature was at45℃and the optimum pH5.0. The enzyme waspurified by a column of DEAE-Cellulose and checked the purification and molecular weightwith the method of SDS-polyacrylamide gel electrophoresis, the molecular weight was77kDa.After many experiments, the crude enzyme-reacted production were separated byAbsorbitive Resins(AB-8), and the crude ginsenoside C-K were separated by the lower press silica gel column to obtain the6.74g100%pure ginsenoside C-K and10g70-80%pure ginsenoside C-K.