Studies On Technology Of Coprinus Comatus Submerged Fermentation And Its Polysaccharide Extraction And Purification

Coprinus comatus is a delicious, nutritional edible and medicinal fungi.Polysaccharides from Coprinus comatus possess a wide variety of physiological activity,such as blood sugar-lowering, immunomodulating, antitumor, etc. Coprinus comatus hasbeen affirmed being one of sixteen valuable and rare edible and medicinal fungi which isnatural, nutritional, health protective by FAO and WHO. As a result, Coprinus comatus isa edible and medicinal fungi which will be widely developed. Technology of Coprinuscomatus, submerged fermentation and its polysaccharide extraction, isolation andpurification were studied in this paper. The main results were as follows:1. By monofactorial and orthogonal test, an optimum medium had been selected. Themedium composition was as follow: 4% maize amylum, 0.2% soybean cake meal, 3%sucrose, 0.1% KH₂PO₄, 0.1% MgSO₄, 0.05% CaSO₄, 0.5% corn steep liquor.2. The suitable shaking culture conditions were as follow: initial pH of medium 6.0,shaking frequency 170r/min, 100mL liquid medium in size 250mL shaking flask,inoculation quantity 10%, fermentative time 6 days. On this condition, the production of15.70g/L biomass and 1.46g/L extracellular polysaccharide were obtained, pH fell from6.05 to 5.43 in course of the fermentation. The result of fermentation experiment in 50Lfermentor showed that the fermentation had been ended up after 120 hours. Theproduction of 15.70g/L biomass and 1.24g/L extracellular polysaccharide were obtained.3. By means monofactorial of test, the extractive conditions of extracellularpolysaccharide were determined as follows: pH of ferment liquid 5.0, addition of 1volume of ethanol, temperature15℃, precipitation time 24h. The extractive efficiency ofextracellular poiysaccharide was 0.18%. By orthogonal test, the extractive conditions ofmycelial water-soluble polysaccharide were determined as follows: water/mycelia=2/1,microwave heating time 3 minutes, extracting temperature 55℃, extracting time 6h. Theextractive efficiency of mycelial polysaccharide was 1.03%.4. Crude polysaccharides were preliminary purified by deproteinization with Sevag'sprocedure and deionization with dialysis. Conditions of deproteinization with Sevag'sprocedure were determined as follows: V_sample:V_{chloroform+n-butanol}=1:1,V_chloroform:V_n-butano=5:1, shaking time was 30 minutes. Efficiency of deproteinization wasabout 69%.5. The partial purified polysaccharides were further graded and purified by DEAEcellulose colume. As a result, three factions in GLP and GMP were obtained, respectively.GLP-â…¡and GMP-â… werethe main and large factions. Then GLP-â…¡and GMP-â… were further graded and purified by Sephadex G-200 column chromatography. The resultshowed that the molecular weight of the two factions were similar which obtained inGLP-â…¡and GMP-â… ,respectively. GLP-â…¡-1 and GLP-â…¡-2 represented 54% and 46% oftotal amount. GMP-â… -1 was a main and large faction, representing 81% of total amount.