Study On Ultrasonic Extraction And Optimization Of Liquid Fermentation Process And Antibacterial Activity Research Of Puffball Polysaccharide

Objective Polysaccharide is one of the most important active ingredients of puffball,which has the functions of bacteriostasis,protection of liver injury,inhibition of tumor cell growth,antioxidation and so on.This study selected Lasiosphaera fenzlii Reich.and Scleroderma polyrhizum(J.F.Gmel.)Pers.as the research object.The ultrasonic extraction technology of RLSP and liquid fermentation technology of Scleroderma polyrhizum were optimized respectively,and the bacteriostatic activity of PSP in vitro was studied.At the same time,the effect of Chinese herbal extract added to fermentation medium on the content of PSP was preliminarily discussed,which provided a reference for further research and development and utilization of these two kinds of puffball.Methods(1)Ultrasound-assisted extraction of RLSP process optimization: Ultrasoundassisted extraction of RLSP is based on single factor experiments.Under the condition of fixed ultrasonic frequency,the extraction time,extraction temperature,liquid-solid ratio are selected as independent variables,and the RLSP extraction rate in solution is The response value was used to investigate the influence of the interaction between the variables on the extraction rate of RLSP by CCD-RSM,and the optimal process parameters of ultrasonic assisted extraction of RLSP were determined.(2)Optimization of nutrient factors in liquid fermentation of Scleroderma polyrhizum: Based on single factor experiment,carbon source,nitrogen source and inorganic salt were selected as independent variables,and PSP content in fermentation broth was response value.CCD-RSM was used to investigate the interaction between the variables.The effect of PSP content on the optimal nutrient factors for Scleroderma polyrhizum.(3)Optimization of liquid fermentation conditions of Scleroderma polyrhizum: Based on single factor experiment and optimization of nutrient factors,the fermentation time,fermentation temperature and inoculum were selected as independent variables,and the PSP content in the fermentation broth was the response value.CCD-RSM was used to investigate the variables.The effect of the interaction on the PSP content determines the optimal fermentation conditions for liquid fermentation.(4)Study on the content of PSP by adding traditional Chinese medicine in fermentation broth: Four kinds of Chinese medicines with different medicinal parts were selected to investigate the effect of water decoction on the content of PSP in fermentation broth by adding different concentration into liquid fermentation medium.(5)In vitro bacteriostatic activity of PSP: Three common pathogenic bacteria were selected and the bacteriostatic effect of PSP solution with different concentration on three pathogenic bacteria was studied by filter paper method.Results(1)The optimum extraction conditions of RLSP were as follows: liquid-solid ratio was 30:1,the extraction time was 53.77 min,the extraction temperature was 58.33 ?,and the extraction rate of RLSP was 3.88%.(2)The optimum nutritional conditions of Scleroderma polyrhizum liquid fermentation medium were glucose 2.8%,peptone 1.8%,magnesium sulfate 0.1%,potassium dihydrogen phosphate 0.1%,and PSP content in fermentation broth 11.03 mg/mL.(3)The optimal culture conditions for PSP liquid fermentation medium were as follows: fermentation temperature 26.6 °C,fermentation time 64.71 h,inoculum volume 9.98%,and PSP content in fermentation broth was 11.73 mg/mL.(4)The addition of Oroxylum indicum and Vitextrifolia L.promoted the content of PSP in the fermentation broth;the low concentration of Morus alba and Viola yedoensis had a weak effect on the PSP content in the fermentation broth,and the high concentration added a strong inhibition effect.(5)PSP has inhibitory effects on staphylococcus aureus and escherichia coli.At high concentrations,staphylococcus aureus has the strongest inhibitory ability,escherichia coli has a weaker inhibitory ability,and its inhibitory capacity decreases with decreasing concentration.For pseudomonas aeruginosa,PSP solution has no inhibitory effect.Conclusions The above three CCD-RSM optimization results were verified respectively,and the verification results were in good agreement with the model prediction values.Therefore,the above three methods can be considered as the ultrasonic extraction process of Lasiosphaera fenzlii and the optimization and prediction of the Scleroderma polyrhizum cell liquid fermentation process;the water extracts of Oroxylum indicum,Vitextrifolia L,Viola yedoensis and Morus alba all have an effect on the change of PSP content in the fermentation broth;the antibacterial ability of PSP increases with the increase of concentration,and the antibacterial ability to Gram-positive bacteria.It is stronger than Gram-negative bacteria.