Studies On Fermentation Culture And Bioactivity Of Ganoderma Lucidum Karst

Ganoderma lucidum Karst is a precious traditional Chinese medicine with a history of more than two thousand years.It belongs to porous bacteria family and grows on the decayed wood of broad-leaved trees.Ganoderma lucidum Karst contains Ganoderma lucidum Karst polysaccharide,Ganoderma lucidum Karst triterpene and other bioactive ingredients,which have the effects of regulating intestinal flora,anti-oxidation,anticancer,lowering blood sugar and blood lipid.However,the quantity of natural Ganoderma lucidum Karst is not enough for commercial use,and the quality of artificial Ganoderma lucidum Karst is unstable and the growth cycle is long.The bioactive ingredients in Ganoderma lucidum Karst can be obtained through fermentation,which has a short cycle and is not easily affected by the external environment.Therefore,more bioactive ingredients related to Ganoderma lucidum Karst can be obtained and the prebiotic function of Ganoderma lucidum Karst can be more effectively played.In this thesis,the fungal tissue collected in Luoshan Scenic Spot of Yantai City was isolated and purified,and the total DNA of the genome was extracted by the fungal total DNA extraction kit,and ITS regional sequence was obtained by PCR amplification.The sequencing analysis showed that the strain was the Ganoderma lucidum Karst of polyporaceae,and the strain was named LS as the starting point.With biomass as the index,the best mycelium growth medium was selected as the basal culture medium from six kinds of medium.Response surface analysis of fermentation time,temperature and p H range were determined by single factor experiment.The optimal fermentation conditions were 28℃,p H=7 and 7 d,mycelium biomass is 1.198 g.The optimal fermentation solution was obtained by the response surface conditions,and the fermentation broth of Ganoderma lucidum Karst mycelium was obtained under the optimal conditions.The contents of key components in the fermentation broth were determined.The first is the change of acidity and the change of reducing sugar content.In a certain fermentation time,the acidity content is proportional to the fermentation time,and the change of reducing sugar content is inversely proportional to the fermentation time.The contents of total polyphenols,total flavonoids and total triterpenes were 0.084mg/m L,0.142 mg/m L and 0.09 mg/m L,respectively,by the national standard method.The basic monosaccharides and the contents of Ganoderic acid A and Ganoderic acid B were detected by HPLC.The contents of glucose,galactose and D-mannose were 201mg/L,238 mg/L and 125 mg/L,respectively.Ganoderic acid A was significantly lower than the level of detection,and the content of Ganoderic acid B was 0.23 mg/L.The inhibitory activity of Ganoderma lucidum Karst mycelium fermentation broth was detected by the bacteriostatic zone experiment.The scavenging rates of DPPH free radical and ABTS free radical in Ganoderma lucidum Karst mycelium fermentation broth was up to 99.81%,80.35%,66.81% and 80.35%,respectively.It was found by MTT assay that Ganoderma lucidum Karst mycelium fermentation broth had an inhibitory effect on the activity of HEP-G2 cells,and the inhibitory ability was positively correlated with the concentration.Ganoderma lucidum Karst mycelium fermentation broth could also promote cell apoptosis,and its ability to promote cell apoptosis was positively correlated with the concentration within the test range.