Study Of Ginsenosidase Hydrolyzing Protopanaxatriol Ginsenosides

The structure of the protopanaxatriol type ginsenoside(PPT)F1 is similar with that of Compound-K(C-K),and it has important application prospect.Ginsenosidase type IV can hydrolyze PPT:it can hydrolyze R1 and Re to Rg1,and further hydrolyze Rgl to F1.This paper is mainly about the prepration of F1 from PPT.To prepare PPT type ginsenoside R1,3.0 kg root of Panax notoginsenoside(BurK.)F.H.Chen was extracted by methanol.After extracted by n-butanol saturated by water,and washed by petroleum ether,241.5g R1 was obtained;the yield was 8.05%.Silica column chromatography was used to purify 45.0 g R1 crude product.2.21 g pure R1 was obtained,the yield was 4.91%,the purity was 98.64%.The enzyme producing microorganism was screened,sp.39 strain was chosen.DEAE was used to purify the ginsenosidase type Ⅳ.The enzyme was performed one single band on the gel,and the molecular mass was 56 kDa.HPLC result proved the SDS-PAGE result.The optimal temperature and pH for this enzyme were 40℃ and 6.0,respectively.The enzyme was stable at pH 3.0-7.0,and below 65℃.To preparae a large amount F1,sp.39 strain was incubated largely.65.0 g PPT(mainly Re and Rg1)was transformed at pH 6.0,40℃ by the enzyme for 24 h.The hydrolysate was firstly purified by macroporous absorption resin,and 37.0 g crude product was obtained,the yield was 56.9%in this step.Then silica column chromatography was used to purify the product.V(chloroform):V(methnol)=9.0:1.0 was used as solvent.6.66 g F1 was obtained which purity was no less than 90%,the yield was 18%;11.19 g Rgl was obtained which purity was no less than 96%,the yield was 30.24%.