A Study On The Polysaccharide Fermenting Techniques And Biological Characteristics Of The Mycelium Of Armillaria Luteo-virens In Qinghai Province

Morphological observations were made on mushroom fruitbodies collected from Qinghai Lake. After carful identification by means of referring to references, they are primiarily designated to be Armillaria luteo-virens. It belongs to Eumycota Basidiomycotina Hymenomcentes Tricholomatales Tricholomataceae Armillaria in classification.After tissue isolation of Armillaria luteo-virens fruitbidies, four strains of Armillaria luteo-virens were obtained among which one was identified to be of experimental strain and this strain was named QHH-1. Systemic research on the biological characteristics and polysaccha- ride fermenting was made in this paper.The results of single factor test of physiological characteristics of Armillaria luteo-virens are as fellows. The glucose and the mannitol are its best carbon source. The grass perchlorate, ammonium chloride, cooked bran juice, ammonium sulfate and beef extract are the best nitrogen source. The potassium dihydrogen phosphate is the best salt for her. The VB₁ is the best growth factor for her. As the value of pH is 7.0, she grown fast. Through the orthogonal test,the optimized medium is as follows: Glucose 0.2g, NH₄Cl 2g, KH₂PO₄ 0.2g, MgSO₄·H₂O 0.2g, FeSO₄ 0.2g,Al₂(SO₄)₃0.2g, Vitamin B₁ 1mg,Vitamin B₂ 1mg , H₂O 1000ml.The experiment which used the methods of phenol-sulfuric acid and Anthrone- sulfuric acid to assay the polysaccharide content involved the color optimization test of glucose, L-rhamnose, galactose, D-fructose and xylose, the test of the maximum absorption wavelength and the stability test. At last ,we chose the glucose as a standard for drawing standard curve of the phenol-sulfuric acid method and Anthrone-sulfuric acid method. And we had the standard curve. When we compared the stability and the ratio of recovery with each other , we found The former is the better for assaying the polysaccharide content of Armillaria luteo-virens.We use the hot water , alkali and acid to extract the polysaccharide of Armillaria luteo-virens. Then optimize the experimental condition. The regression equation used the hot water is y=10.38-0.31 x₁-0.51 x₂-0.18 x₃ + 0.25 x₂²-0.88 x₃², and its optimized experimental condition is the solid-liquid ratio 1:10 , the extraction time 1 hour, the extraction temperature 79℃.The regression equation used the alkali is y=10.32 +1.28 x₁+0.38 x₂ -1.98 x₂², and its optimized experimental condition is the value of pH 11.0 , the extraction time 1.16 hour , the extraction temperature 71.3℃. The regression equation used the acid is y=9.42 + 1.16x₃ - 2.1 x₁ x₂+0.80 x₁²-1.09 x₃², and its optimized experimental condition is the value of pH5.0 , the extraction time 1.5 hour, the extraction temperature 57.6℃.When we compared with each other, we found the method used the acid is the best for extracting the polysaccharide of Armillaria luteo-virens.Through the uniform design and regression analysis, the regression equation of polysaccharide fermentation about Armillaria luteo-virens mycelium is y= 13.747-0.27 x₂-0.87x₃ - 0.562 x₄ + 0.062 x₅ - 0.065 x₆.The optimized experimental condition of polysaccharide fermenting of Armillaria luteo-virens is the sucrose 5.0g , (NH₄)₂C₂O₄ 0.5g , Beef extract 0.5g, K₂HPO₄ 3.0g , Vitamin B1 0.05mg. We elementarily summarize the process of polysaccharide fermentation and extraction.The process of polysaccharide fermentation is culture→one-level seed→two-level seed→three-level seed→liquid fermentation→polysaccharide extraction. The process of polysaccharide extraction is mycelium→desiccate→comminute→extract the polysaccharide use the acid→concentrate,alcohol precipitation,wash→desiccate→crude polysaccharide.