Study On Microbial Transformation Of Baicalin And Wogonoside In Scutellaria Baicalensis Georgi

Objective:In This study is microbial transformation of Scutellaria baicalensis Georgi by the biological transformation technology,to provide experimental data for further development and utilization of Radix scutellariae.Methods:1.We obtained beta glucuronic acid in to hydrolysis of flavonoid glycosides bond strain of Capable of hydrolyzing by screening.2.Wedesigned L₉?3⁴?orthogonaltestbyinvestigatingthesinglefactorexperiment,and optimizedfermentation method of solid and liquid of Baicalin.3.To determine the optimal liquid fermentation process of Scutellaria baicalensis,we investigated the influence carbon source,nitrogen source,inorganic salt,different concentration of nitrogen source and inorganic salt on baicalin liquid fermentation process;and also observed the fermentation time,the sample loading,speed,pH value,solid-liquid ratio and other factors.4.To establish a method of the separation of fermentation conversion product extracted from Scutellaria baicalensis fermentation samples,we used the method of Solvent extraction to investigate conditions of fermentation sample,and optimized macroporous adsorption resin.Transformation products using TLC,HPLCand¹H-NMR,¹³C-NMR,LC-MSidentificationofRadix scutellariae.Results:1.Bacillus natto could hydrolyze of baicalin and wogonoside in beta glucuronic acid to generate aglycone baicalein and wogonin.2.We compared the liquid with solid fermentation technology,and determined the solid fermentation method of more suitable for the fermentation of Radix Scutellariae,and obtained optimal fermentation conditions:After cultured 12h at 180 r/min,Bacillus natto were inoculated into the crushing 40 objective in Scutellaria baicalensis Georgi,liquid-solid ratio 3:1,30%inoculation quantity,culture temperature of 37?,fermentation time 7d.After fermentation,the rate conversion of Baicalin was 96.5%,the rate of conversion of wogonoside was 98.6%in Scutellaria baicalensis sample.3.Extraction of Scutellaria baicalensis Georgi fermentation samples by solvent method,extract of baicalein extract rate was 79%,wogonin rate was 76%.By the experiment of optimized get the macroporous resin separation and purification conditions:the macroporous adsorption resin D101,column on the speed of 2BV/h,the column liquid concentration(2.47mg·mL^-1 of baicalein,wogonin 0.57 mg·mL^-1),60%ethanol elution of baicalein and 80%ethanol elution of wogonin,elution speed 3BV/h,diameter height ratio of 1:7,the adsorption capacity for 32.6mg/g.It identified baicalein and wogonin of 2 conversion products of fermentation of Radix Scutellariae of the TLC,HPLC and ¹H-NMR,¹³C-NMR,LC-MS and separated 97%purity of baicalein and wogonin purity of 98.6%.Conclusion:According to the above test results,can be identified of Bacillus natto as fermentation strains in Scutellaria baicalensis Georgi,that has high efficiency,specific characteristics.Useing of solid fermentation method of fermentation Scutellaria baicalensis Georgi,enhance the transformation product quantity,the combined effect of solvent extraction method and macroporous adsorption resin,that separated and purificated to obtain conversion product of baicalein,wogonin.We determined the method of Scutellaria baicalensis Georgi biotransformation and extraction and separation,this method is stable and feasible,convenient conditions,and enhanced the scutellaria root physiological activity and medicinal value.Our study plays a certain role in promoting to continue the development and utilization of Scutellaria baicalensis Georgi by microorganisms transformation.