Study On The Effect Of Liquid Fermentation Of Cordyceps Militaris And Its Active Extracts On Intestinal And Physiology Of Mice

Cordyceps militaris belongs to the Fungi community,the Fungus gate,the Ascomycete subphylum,the Nucleobacteria,the genus Lycoris,and the genus Helminthaceae.It is rich in physiological activities such as cordycepin,adenosine,cordyceps polysaccharide and cordycepic acid,and has physiological functions such as regulating immunity,anti-tumor,anti-fungal,anti-viral,anti-leukemia,blood sugar lowering and blood fat lowering.At present,the products on the market are mainly based on fruiting bodies,and the fruiting bodies are faced with the problem of loss of nutrients and active ingredients in the long-term preservation process.The liquid fermentation of Cordyceps militaris has a short production cycle and is easy to control automatically.The active ingredients in the fermentation broth are abundant.The extraction and deep processing of the active constituents in the mycelium and fermentation broth of Cordyceps militaris can enhance the preservation of the active components of Cordyceps militaris and the absorption of the body.More and more studies have found that the intestinal flora is inextricably linked to the health of the body.In this study,we studied the effects of Cordyceps militaris on intestinal microbes by fermenting,extracting active ingredients,and then using mice as animal models.The main research contents of this thesis are as follows:1.Optimization of liquid fermentation medium and conditions for Cordyceps militaris.Through single factor experiment and orthogonal experiment,the optimal medium is:D-fructose 4%,beef extract 1.5%,calcium chloride 0.1%,VB₁ 10mg/L;the fermentation conditions are:fermentation time 9 days,temperature 25?,The rotation speed is 120 r/min,the initial pH is 5,the liquid volume is 100 ml,and the inoculum amount is 10%.Finally,the optimization results were verified,and the mycelium fermentation of Cordyceps militaris was as high as 18.838 g/L.2.The extraction process of active ingredients in mycelium was optimized,and the extraction conditions were determined:the temperature is 100?,the ratio of material to liquid is 1:20,and the extraction rate is 10.56%.The extract was mixed with the extracted fermentation broth to synthesize Cordyceps polysaccharide by using a complex enzyme?cellulase and hemicellulase each 20,000 U/g polysaccharide?,and hydrolyzed at pH 4.5 and a reaction temperature of 45?.The oligosaccharide production was the highest at min.After centrifugation?speed 10000 r/min,feed amount 4 L/min?separation can effectively obtain a relatively clear fermentation broth.The ultrafiltration membrane and the nanofiltration membrane filter the yield of the effective substance,wherein the ultrafiltration conditions are:pressure 2 MPa,flow rate60 L/h;nanofiltration conditions:pressure 3 MPa,flow rate 30 L/h.Finally,the concentration is 55? and the vacuum degree is 0.08,so that the dry matter concentration of the material reaches 180 g/L.The powder particles obtained by spray drying are uniform,the water content is low,and the dry operation parameters are:the inlet air temperature is 140?,and the air is discharged.The temperature is 85?.3.Effect of Cordyceps militaris powder on intestinal and physiology of mice.The mice with the intestinal flora disorder model were constructed.After successful modeling,different doses of Cordyceps militaris extract were administered,and water,oligomannose and normal were used as the control group,and the administration time was one month.The body weight and food intake of the experimental group were recorded,and it was found that the body weight of the mice in the administration group was significantly reduced compared with the control group.The feces were collected and diluted and coated at each time point to investigate the changes of live bacteria of Enterobacter,Enterococcus,Lactobacillus and Bifidobacteria.The results showed that the extract of Cordyceps militaris had a certain regulation effect on the number of viable bacteria in the feces.After the end of the administration,the serum of the mice was taken for detection,and it was found that the extract of Cordyceps militaris had the effects of improving blood lipids,protecting the liver and the heart.The mouse feces,colon contents and ileal contents were taken,and the total DNA was extracted for 16S rDNA sequencing analysis.It was found that the extract of Cordyceps militaris had a certain ability to regulate the abundance,diversity and obesity-related flora of the intestinal flora.