Screening And Preliminary Analysis Of Edible And Medicinal Mushrooms Based On Inhibition Of Non-enzymatic Glycosylation

Non-enzymatic glycosylation (NEG) reaction of protein is one of the most importantfactors in the occurrence and development of diabetes, and closely related with a variety ofchronic diabetic complications. Therefore inhibition of NEG reaction is an effective way toprevent and treat diabetes and its complications. The natural bioactive compounds in edibleand medicinal mushrooms are main reasons for their pharmaceutical functions. The study ofthese bioactive compounds is important to evaluate the application of edible and medicinalmushrooms, pharmaceutical functions and mechanisms. In this study, various edible andmedicinal mushrooms were screened by their inhibitory rate of NEG reaction undersubmerged fermentation and the high inhibitory rate mushroom was selected to analyze thestructure and function of bioactive compounds. The main result of this study was described asfollows:1. The inhibitory rate of23screened mushrooms in vitro were analyzed and resultsindicate that only4of them, Lentinus edodes, Pleurotus eryngii, Tremella fuciformis andAgaricus blazei murrill, possess high inhibitory in mycelia comparing to culture broth. Thebioactive compounds in other23mushrooms were distributed in culture broth and3mushrooms, Armillaria mellea, Morehella esculenta and Hericium erinaceus were selectedfor further research.2. A. mellea fermentation broth was separated by two-step ethanol precipitation, ethylacetate extraction and other steps, and four compounds possessed high inhibitory activity. Thepolysacchrade compouds, AMP2-I, were purified by DEAE ion-exchange column and gelcolumn and mainly composed of glucose, mannose, arabinose, galactose, xylose and fucose ina molar ratio of28.69:1.00:0.50:0.40:0.38:0.06. The process of AMP2-I by partial acidhydrolysis significantly enhanced the inhitory rate to NEG reaction, which reached89.70%in2mg·mL-1.3. Four ingredients were purified from the M. esculenta fermentation broth, and thehighest bioactive compounds, ME3, was obtained from the ethyl acetate extract, which wasfurther seperated7compounds. However, the inhibitory rates of these7compounds weresignificantly reduced compared with ME3. To identify the main bioactive compound,6different compounds were mixed and inhibitory rates of mixtures were analyzed. The resultindicates that3or more compounds were responsible for NEG reaction and ME3-5was themain bioacitive compound. Other3compounds enhanced the inhibitory rate of ME3-5, whichreached88.59%in2mg·mL-1.4. The biological active component inhibiting NEG reaction in H. erinaceus fermentationbroth was purified by the two-step ethanol precipitation, ethyl acetate extraction, C18chromatography and HPLC. The obtained high inhibitory single substance (HE35) had clearabsorption peaks under the wavelength254nm and300nm. After analyzing by LC-MS, therelative molecular mass was294. In vitro experiment, the inhibitory rate of HE35to NEGreaction under2mg·mL-1was92.17%and IC50was0.65mg·mL-1. The result of massspectrometry and NMR analysis of HE35inferred that its formaula was C20H22O2. Good inhibitory activity of a variety of edible and medicinal mushrooms demonstratedthe species’ biodiversity on NEG reaction. Bioactive compounds not only containpolysaccharide and single biological active substance, but also contain several compoundscomplicated mixing some substances, and also demonstrated materials biodiversity on NEGreaction.