Fermentation Of Coprinus Comatus With Herbs And The Activity Component Of The Fermented Broth

Coprinus Comatus is a kind of medible and medical fungus, its fermentation broth hasthe hypoglycemic function. Polysaccharides from Coprinus Comatus possess a wide varietyof physiological activity, such as anti-cancer, antitumor, immune modulating, reducing highconsistency of blood glucose, etc. The work in this dissertation was adding different Chinesemedicine into the basic medium cultured by C. Comatus in order to enhance its hypoglycemicactivity. The effects of some Chinese medicines on C. Comatus growth and exopolysaccharideexcretion were investigated, the change of active compounds of C. Comatus cultured with M.charantia L. was syudied, and the invitro activity of the fermentation broth was tested. Thebioactivity of active compounds was also explored.Some works were carried about effects of nine kinds of Chinese medicines on biomassand crude exopolysaccharide of Coprinus comatus in submerged fermentation. The resultshowed: M. charantia L., G. pentaphyllum, Lily, Buckwheat, R. S. miltiorrhizae, Ginseng andA. konjak could improve the growth of C. comatus and stimulate the secretion of crudeexopolysaccharide in a certain dosage, but A. menbranaceus and P. odoratum inhibited thefermentation of C. comatus greatly. It could be concluded, by adding medicines in differentdosages, that the optimum dosage of M. charantia L., G. pentaphyllum, Lily, Buckwheat, R. S.miltiorrhizae and A. konjak was 0.5%, and that of Ginseng was 1.5%. The test of invitroactivity showed that the inhibitory rate of the fermentation broth on α-glucosidase and formationof AGEs was enhanced remarkably when M. charantia L. was added to the medium beforefermentation.Through experiments, the optimum pH before the fermentation of C. comatus culturedwith M. charantia L. was 6.3, and the optimum dosage of M. charantia L. was 0.75%. Itsinhibitory rate on formation of AGEs reached (96.9±0.3)% under these conditions. After72-houurs fermentation in 30L fermentor, productivity of 0.8680g/100mL biomass and 0.6129g/100mL crude exopolysaccharide was obtained.C. comatus was submerge-cultured with the medium containing M. charantia L.. Thoughthere was no new exopolysaccharide component produced by the addition of M. charantia L.,the yield of exopolysaccharide was improved and the relative content of exopolysaccharidecomponents was changed. After separated by DEAE-cellulose column and Superdex 200column, the two main polysaccharide, CMP-ⅠA and CMP-ⅡB, were obtained. The averagemolecule weight of the two fractions was estimated by HPLC to be 23455Da and 20267Darespectively. The structure of CMP-ⅠA was elucidated by glycosyl residue compositionanalyses, IR spectroscopy, UV spectroscopy and NMR spectroscopy. The sugar componentswere arabinose, xylose, mannose, glucose, and galactose, and the ratio was 4︰16︰4︰1︰65.The main chain of CMP-ⅠA would be composed by -β-D-Gal(1→6)-, and β-D-Xyl,arabinose, mannose, glucose and acetyl would be in the side chain.The fermentation broth of C. comatus cultured with M. charantia L. was separated by themacroporous absorption resin, and the inhibitor, named EC, was obtained. The half-inhibitoryconcentrations of exopolysaccharide and EC on α-glucosidase were about 2mg/mL and1.5mg/mL. Exopolysaccharide belongs to competitive inhibitor, and EC belongs tononcompetitive inhibitor. The Ki value of exopolysaccharide and EC were 1.14mg/mL and1.20mg/mL respectively. The half-inhibitory dosage of exopolysaccharide on formation ofAGEs was 6 mg/mL, but that of EC was only 0.7mg/mL.