Study On The Biotransformation And Separation And Purification Of Panax Notoginseng Root And Leaf Extract

Panax notoginseng is a valuable resource of Traditional Chinese Medicine in China.The roots,stems and leaves of P.notoginseng can be used as medicines,and saponins are the main components for its medicinal use.In this work,the extracts of P.notoginseng root and leaf were biotransformed and purified,the high content sponins Rbl,Rd and Fd were transformed into rare saponins F1,CK elso by microbial enzymes,and enzymes were separated and purified.(1)The saponins in the extracts of P.notoginseng root and leaf were identified by HPLC and LC-MS.Aspergillus niger,which can specifically transform the major saponins in the extracts of P.notoginseng root and P.notoginseng leaf into rare saponins F1,CK and saponin ⅩⅢ,was screened from the strains preserved by our group.(2)The factors influencing the transformation of P.notoginseng root extracts into rare saponins F1 and CK by Aspergillus niger enzyme solution were studied:transformation temperature,pH,enzyme concentration and culture time.The results showed that after 4 days of cultivation,the conversion of P.notoginseng root extracts(1%)was carried out for 72 h,the conversion rates of saponin R1,Re,Rg1,was 88.6%,91.0%,71.8%,and the concentration of F1 was 2.82 mg/mL,the yield was 84.7%.After 5 days of solid cultivation of Aspergillus niger,the extracts of P.notoginseng root(1%)was transformed 72 h.The conversion rate of saponin Rb1 was 99.8%.The conversion rate of saponin Rd was 84.3%.The highest concentration of Rd was 3.16 mg/mL at 0.5 h(the optimum time for obtaining the maximum amount of Rd),and the highest concentration of saponin F2 was 1.69 mg/mL at 6 h(the optimum time for obtaining the maximum amount of F2).After 72 h,the concentration of saponin CK was 2.91 mg/mL,and the yield was 85.8%.(3)The factors influencing the transformation of P.notoginseng leaf extracts into saponins ⅩⅢ and CK by Aspergillus niger enzyme solution were studied,including transformation temperature,pH,enzyme concentration and bacterial culture time.The results showed that after 5 days of cultivation,the conversion of P.notoginseng leaf extracts(2%)was carried out for 108 h,the conversion rate of saponin Fe was 82.9%,the concentration of Mc was 1.64 mg/mL,the yield was 81.9%,the conversion rate of saponin Fd was 89.0%,and the concentration of ⅩⅢwas 2.80 mg/mL,the yield was 81.3%.This work is the first time to prepared saponinⅩⅢ from the extracts of P.notoginseng leaf.(4)The static adsorption and desorption experiments on macroporous adsorption resin showed that the macroporous resin D302 had better separation effect on saponins F1,CK and ⅩⅢ After purification by macroporous resin,saponin F1 3.88 g was obtained from the extracts of 20 g P.notoginseng root with a recovery of 69.5%.Saponin CK 3.94 g was obtained with a recovery of 68.4%.Saponin ⅩⅢ 2.01 g was obtained from 20 g P.notoginseng leaf extracts with a recovery of 73.9%.Saponin CK 1.64 g was obtained with a recovery of 69.2%.(5)Salting out method was used to separate and purify the enzyme protein from Aspergillus niger.It was infered that the enzymatic protein producing saponin F1 was ginsenoside type Ⅳ enzyme,the enzymatic protein producing saponin CK was ginsenoside type Ⅰ enzyme,and the enzyme producing saponin ⅩⅢ was ginsenoside type Ⅱ enzyme.