Study On The Effect Of Lactic Acid Bacteria Fermentation On Ginsenoside And Its Antitumor Activity

Ginseng is a perennial herb of Araliaceae,traditional Chinese herbal medicines,was identified as the new resources of food in 2012.The main active ingredients of ginseng are rare ginsenosides.At present,most researches were about fungi and the conversion of a single ginsenoside in MRS medium.These were risks of the food safety and health.The application in food processing was limited.In this study,skim milk was as substrate,ginseng crude extract was as substrate,the effect of lactic acid bacteria fermentation on ginsenosides and its antitumor were studied,making a the foundation for the development of ginseng fermented food,Through the exploration and the experiment of HPLC,the simultaneous determination seven kinds ginsenosides method was built.The methodological evaluation showed that this method was reliable.The sample separations were achieved with a constant flow rate of 0.8 m L/min at 35 oC,and the detection wavelength was set at 203 nm.The mobile phase consisted of solvent A(acetonitrile)and solvent B(water)using a gradient elution of 18-23% A at 0-10 min,23-44% A at 10-30 min,44-68% A at 30-38 min,68% A at 38-45 min,68-100% A at 45-55 min,100% A at 55-60 min,100-18% A at 60-63 min.Ginseng crude extract was used as the research object,the skim milk was used as the substrate environment.The transformation of ginsenosides by five lactic acid bacteria was carried out to obtain the four kinds of protopanaxadiol saponins The transformation pathway was Rd?Rg3?Rh2 and Rd?CK.Fermentation.Rd content decreased,Rg3,Rh2 and CK content increased.Lactic acid bacteria promoted protopanaxadiol ginsenoside transformation,the degree of dextran was increased.The transformation pathway of the original ginseng tartrate saponin is Re?Rh1?PPT.Glycosylation of protopanaxatriol ginsenoside with different lactic acid bacteria was different.Lactobacillus rhamnosus had better effect on transformation of protopanaxadiol saponin in 72 h.culture conditions for the transformation of ginsenoside by Lactobacillus acidophilus was optimized.The single factor experiment showed that the optimum fermentation temperature was 37 ?,the initial p H was 6.8 and the inoculation amount was 3%.The orthogonal experiment showed that the optimal level of factor was A2,B2C2,namely the fermentation temperature was 37 ?,inoculation amount was 3%,the initial p H was 6.8.Add glucose,maltose,sucrose,fructose,sorbitol,soybean oligosaccharides,inulin to the culture medium,not to add a carbon source as the control group.For protopanaxadiol ginsenosides,it was found that adding glucose significantly increased the content of rare ginsenoside.The content of Rg3 was 0.81408?g/20?L,increased by 3.211%.compared with control.The content of Rh2 was 0.43122?g/20?L,increased by 9.002% compared with control;the content of CK is 0.46913?g/20?L,increased by 12.980% compared with control.For protopanaxtriol ginsenoside,the content of Rh1 had a decrease,which indicates that the addition of carbon source promoted transformation of protopanaxtriol ginsenoside,promoted the degree of hydrolysis of glycosyl at C-6 of Rh1.Through the analysis of transformation of ginsenoside by Lactobacillus rhamnosus in72 h,the p H was reduced from 6.823 to 3.973 at 0-36 h.The p H was stable at 36-72 h,and was 3.823 at the fermentation end point.Ginseng total saponin showed a downward trend,in 24-48 h decreased flat,total saponin content decreased significantly in 0-12 h and 48-60 h.After 72 h of fermentation,the total saponin content in the sample decreased from 0.1797mg/0.1m L to 0.1103mg/0.1m L,a decrease of 38.62%.The contents of ginsenoside in the protopanaxadiol saponins showed a decreasing trend,and the content of rare saponins increased.In the 0-12 h fermentation process,the viscosity of the fermentation broth decreased;12-36 h,the viscosity increased;36-60 h,the viscosity remained basically was unchanged;60-72 h,the viscosity decreased.Polysaccharide content increased from 0-36 h,from the initial 1.2403mg/0.1m L to 1.4566mg/0.1m L,an increase of 17.4%,36-48 h content remained unchanged,48h-60 h content decreased,60-72 h content of little change.The change trend of solution viscosity and sugar content was basically the same,there was a positive correlation between solution viscosity and sugar content.The DPPH clearance rate of the samples without Lactobacillus rhamnosus was lower than that of the fermented samples.?-glucosidase activity was decreased after fermentation,decreased more obvious in 0-12 h,activity remained essentially unchanged in 12-72 h.The inhibition rate of Hep G2 cells of the samples increased with the concentration of the test substance before and after the fermentation.When the concentration of the test substance was the same,the inhibition rate of Hep G2 cells of fermented samples by Lactobacillus rhamnosus was higher than that of the unfermented samples,indicating that the inhibitory activity against Hep G2 cells of the conversion of ginsenoside fermented by Lactobacillus rhamnosus was increased.