Studies On The Extracting Technology Of Polysaccharide And Pharmacological Effects In Morchella Crassipes

In this paper, the experiment chooses the Qinghai-Tibet Plateau Morchellacrassipes as the research material, by liquid fermentation technology to obtainfermentation liquid and mycelium. To study the element content in fruiting body andmycelium of wild Morchella crassipes; With fresh fermented liquid and mycelium asraw materials, respectively for polysaccharide extraction of single factor experimentand orthogonal experiment design, studied the extracellular polysaccharide andmycelium polysaccharide extraction of optimum technological conditions.Weresearch the Pharmacological effects of fermented liquid, mycelium, extracellularpolysaccharide and mycelium polysaccharide, in order to provide theoreticalfoundation and scientific basis for further research to fermentation products andpolysaccharide.Main research contents and results are shown below:1. This paper adopts the inductively coupled plasma-mass spectroscopy methodto study the Mg、K、Ca、Cr、Cu、Zn、As、Se、Mo、Cd、Hg and Pb this12elementscontent in entity and mycelium of wild Morchella crassipes from Qinghai-TibetPlateau. Experimental results show that,enrichment of fruiting body and myceliumof Mg、Zn、Cu、Pb、Hg and Se this six kinds of elements content is quite,which candevelop mycelium by liquid fermentatin instead of fruiting body. The entity andmycelium of concentration of heavy metal Pb, Cd, As and Hg content are within thescope of the food safety standards set by the state, in line with the national foodsafety regulations, people are safe to eat.2. With fresh fermented liquid and mycelium as raw materials, in order to getthe yield of extracellular polysaccharide and mycelium polysaccharide as the index,respectively to adopts four-factors at three levels orthogonal research design whichis determined by single factor experiments method.(1)In the research, the four factors: concentration ration, temperature, alcoholconcentration and pH, was investigated to study the influences on the content ofextracellular polysaccharide. The result shows that the optimal extraction processshould be achieved by concentration ration of1:5, concentrated temperature of50℃,sank alcohol concentration is95%, pH of fermented liquid was changed to6.0.Through the proof test, the average extracellular polysaccharide content is1.275 mg·mL-1,which proves the process condition is stable and reliable.(2)Assisted by ultrasonic extraction method to extract mycelium polysaccharide,the four factors: the ultrasonic treatment temperature, solid-liquid ratio, time and thenumber of processing, the result shows that the optimal extraction process should beachieved by solid-liquid ratio of1:20, ultrasonic treatment temperature of70℃,ultrasonic treatment time of20minute, extraction2times. Through the proof test,the average mycelium polysaccharides content is56.761mg·g-1, which proves theprocess condition is stable and reliable.3. Research on the anti-fatigue effect in Morchella crassipesWith the fresh fermented liquid and mycelium suspension for the reagent,through the weight loading swimming experiment of mice to study the swimmingexhaustion time and immune index(spleen and thymus index). Through the freeswimming experiment of mice and the related anti-fatigue biochemical indicators(LD, BUN, liver glycogen and muscle glycogen) to studied the anti-fatigue effect inMorchella crassipes. Experimental results show that the fermented liquid andmycelium suspension can significantly extend the exhaustion of mice swimmingtime（P<0.01）, improve mice spleen and thymus index, reduce the serum LD（P<0.01）and serum BUN concentration, increase myocardial glycogen（P<0.01）,quadriceps glycogen and the content of hepatic glycogen（P<0.01）, therefore, goodanti-fatigue function was discovered in Morchella crassipes.4. Research on the anti-aging effect in Morchella crassipes(1)With the fresh fermented liquid and mycelium for the reagent, through thefree swimming experiment of mice and the related anti-aging biochemical indicators(SOD in mice serum superoxide dismutase, myocardial superoxide dismutase SOD,serum hydrogen peroxidase CAT and the liver hydrogen peroxidase CAT) andimmune index (spleen index and thymus index) to study the anti-aging effect inMorchella crassipes. Experimental results show that the fermented liquid andmycelium suspension can significantly improve the serum SOD （P<0.05）andmyocardial SOD （P<0.01）vitality in mice, serum and liver CAT vitality, improvemice spleen and thymus index, therefore, good anti-aging function was discovered inMorchella crassipes.(2)Male fruit flies as the research object, used extracellular polysaccharide,mycelium polysaccharide, fermented liquid and mycelium suspension for the reagent,through the life of flies experiment to study the anti-aging effects of Morchellacrassipes.The experimental results show that they can prolong the average lifeexpectancy of males, half the time of death, and the highest life expectancy, has the function of anti-aging.5. Research on the anti-stress effect in Morchella crassipesWith the fresh fermented liquid and mycelium suspension for the reagent,through resistant to high temperature of45℃, resistant to low temperature of-5℃and anti-hypoxia experiment (atmospheric oxygen) to study the anti-stressfunction in Morchella crassipes. The experimental results show that, In resistance tohigh temperature experiment, the fresh fermented liquid can obviously prolongsurvival time of female mice in high temperature environment（P<0.05）.Myceliumsuspension both male and female mice can extend the survival time, and it is betterto female mice. In resistance to low temperature experiments, the fresh fermentedliquid and mycelium suspension can significantly prolong the survival time of micein low temperature environment, prove its resistance to low temperature effect isbetter.In atmospheric oxygen experiment, the fresh fermented liquid oxygenresistance effect is not obvious, mycelium suspension is better（P<0.01）, better thanthat of the positive drug three rhodioal, explain the anti-stress function.6. Research on the anti-microbial effect of fresh fermented liquid fromMorchella crassipes in vitroBased on the fresh fermented liquid from Morchella crassipes for the reagent,by Mclntosh turbidimetric instrument to determine the bacterial concentration, usingfilter paper diffusion method, in the circle of inhibiting bacterium and the minimumbacteriostasis concentration as an index, respectively study the anti-microbial effectfrom fermented liquid to six kinds of strains. The experimental results show that thefermented liquid to six kinds of strains has different degree of anti-microbial effect,the obvious anti-microbial effect including on Escherichia coli, Staphyloccocusaureus, Sarcina maxima and Bacillus subtilis.