The Research On Extraction, Purification And Application About Trehalose That Was Collected From Lepista Nuda Liquid Fermentation

Lepista nuda was belong to Agaricales of Tricholomtaceae.Lepista nuda not only has edible value,but also has officinal value.The research of experiment on collecting fruit body by woodland culture of lepista nuda has been test.For the liquid fermnentation of Lepista nuda and it's extraction of active component, there are few reaserch about this topic.In domestic fungus,more than 70%of lib-sug and lib-sugar alcohol are made up of one or two kinds of sugar.The main component in Lepista nuda is Trehalose,and it's content is rather great.The Trehalose can be used to study food,cosmetic and medicine.In our country,the domestic fungus are very plenteous,so developing the Trehalose have great advantage by domestic fungus. In this study,considering the Lepista nuda-hyphal as my studying object,the extraction,purification, assessment and application for the Trehalose of the Lepista nuda liquid fermentation hyphostroma were studied in this exprement.The result that was investigated are following:1.According to the dry weight of hyphostroma and cont of trehalose confirm the medium's composition in the lepista nuda liquid fermentation.The experiment shows that the most compatible nitrogen source is yeast extract,and the carbon source is glucose,and the inital pH is 7.2.The study of best extract craft parameter for the Lepista nuda hyphostroma.Through single-factor and the orthogonal design,which is to three factors and three levels,I study the extracting time and temperature,and study the effect,which alcohol saturation impact the extraction of Trehalose from Lepista nuda.The best extracting parameter is A₂B₁C₂ The data I have studied as follows:time is 1h;the temperature is70℃;concentration of ethanol is 30%.The cont of Trehalose I gained is 12.92%,and protein level is 0.62%.3.Purification of Trehalose.And I adopted the Sevag to deproteinize.In this experiment,I found as follows:in Sevag,when the ratio of sample volume between Trehalose and chloroform n-butanol is different,it does't only effect rate of deproteinization,but also effect the lossing degree of Trehalose.I sumed up the conclusion from the materials above as follows:the best volume ratio between the Trehalose, chloroform and n-butanol is 5:1:0.25.According determining protein level,I choose the time of deproteinization is twice.After twice deproteinization,the protein level in the Trehalose solution is 0.3%. Using four factors and three levels for activated carbon decolorization,after the addition of the Trehalose solution,the optimal craft of decaloration by activated carbon was:adjusting pH to 4,put in 3%activated carbon,agitating 10min in 60℃.The bleaching rate is 72.04%;cont of Trehalose is 8.06%.The best craft condition is 1mL/min;pH is 6.In this time,the bleaching rate is more than 90%,and desalting rate is more than 97%.4.Using TLC to accredit Trehalose.spreading agent:n-butanol:Pyr:water=15:3:2;Sample is 1.0ul.The liquor rate:vitriol:Meoh =1:4.heat it under 105℃in baking oven in order to colorate.The ratio of crystallization is 2.24%,and the content of Trehalose is 92.56%.5.The study shows that the Trehalose has the function of anti- freezing and anti-fatigue.In the part of anti-freezing,I observe the color,and smell and water quality of Fish wasteful in different time.In the part of anti-fatigue,the Trehalose can have the influence on the Comparison of time to grasp the rope of mice, Comparison of time to swim of mice,serum urea nitrogen and hepatic glycogen.