Research On Domestication And Cultivation Of Inonotus Hispidus And Antitumor Activity Of Its Solid Fermentation Powder And Developing New Product With It

Inonotus hispidus is an annual large medicinal fungi, and is distributed in the north area in China, it mainly grows on mulberry, while the one which distribute in Taonan Jilin Province grows the Ulmus macrocarpa Hance var. mongolica Liou et Li still has no research. In this paper, the two parts of it were studied.The part of summary introduces the cultivation techniques, chemical constituents and pharmacological effects of I.hispidus briefly.Part of the test is into six parts, put 8 kinds of fungi together which known as “Sanghuang”, and do some experiments in a comparative research, and take the I. hispidus do some experiments, such as the domestication, cultivation and solid fermentation of it, and also have some researches on the nutriention, safety and the antitumor activity of fermentation bacteria powder, and selected the better fermentation powder to developed new productions.The first chapter of the test section investigated I. hispidus might a kind of “Sanghuang” by experiment and demonstration, and do experiment on the polymorphism of 8 samples. Used the SLAF-seq technique to analysis the polymorphism of 8 samples known as “Sanghuang” in China, which called Phellinus igniarius, I. hispidus, Phellinus yamanoi, Inonotus xeranticus, Inonotus vaninii, Inonotus linteus, Inonotus sanghuang and Fomitopsis pinicola. The results show that the samples called I. hispidus and P. igniarius have a closer relationship. Comprehensive analysis the common SLAF labels of 8 samples and the picture recorded in ancient book which called “Da guan ben cao”, the fungi which called I. hispidus is more closer to the description of “Sanghuang” which recording in ancient book.The second part of the test section did experiment about domestication and cultivation of I. hispidus. Effect of different carbon source, nitrogen source, initial pH and temperature on mycelial growth of I. hispidus on the solid culture condition were investigated by orthogonal experiments. The optimal cultivation conditions for mycelial growth were 30℃ for temperature, 6.5 for initial pH, xylose for the best carbon source and the yeast extract power for the best nitrogen source. The natural substrates test indicated that corn and sorghum significantly enhanced the initial growth and continuously growth. After selection, the best substrate formulation for the mycelial growth is composed by sawdust 78%, rice husk 2%, corn flour 17%, sugar 1%, lime 1%, gypsum 1% and water 60- 62%, which supported the mycelial growth to fill incubation well within 28 days and immediately started to produce fruitbodies.The third part of the test section did the experiments about the antitumor studied the inhibition and the mechanism of the fermentation powder of I. hispidus on H22 bearing mice. The H22 transplanted models of mice were established by H22 ascites tumor. Ninety mice were divided into nine groups as control, model, cytoxan(CTX, 20mg/kg) positive control, low-, mid-, high-dose(500, 750, and 1000 mg/kg) of corn fermentation powder of I. hispidus, and low-, mid-, high-dose(500, 750, and 1000 mg/kg) of sorghum fermentation powder of I. hispidus. On the day after being inoculated with H22 tumor cells, mice were given fermentation powder of I. hispidus once daily for consecutive 11 d. And then the increase of body weight, tumor weight, the tumor inhibitory rate, spleen, thymus, and liver indexes were calculated; and made the tumor of H22 bearing mice with organization epidemiological research.The activities of MDA, SOD, CAT, and the contents of GSH-Px in liver homogenates and blood serum, as well as the number of IL-2, VEGF, and IFN-γ in blood serum were detected. The tumor growth inhibitory rate of high-dose of sorghum fermentation powder of I. hispidus was 56.12%. The treatment with fermentation powder of I. hispidus significantly increased the body weight, spleen index, thymus, and liver indexes. The research of organization epidemiological research showed that the necrosis in H22 cells was induced and can promote the hepatic SOD, CAT, GSH-Px activities, and decreased the MDA in liver homogenates and blood serum. Elevated the serum levels of IL-2, IFN-γ, and reduced the VEGF level. Fermentation powder of I. hispidus had an antitumor effect on H22 bearing mice, and the possible mechanisms may be due to its antioxidant activity, improved immune function, and anti-angiogenetic action.The fourth part of the test section did the comparative analysis experiment which was conducted about the nutritional components of I. hispidus and its fermentation powder. Through some experiments about the 3 samples, the crude protein, crude fat, inorganic elements, 16 kinds of amino acids and heavy metal content were used to determinate, and had a comprehensive analysis about the 3 samples. The results showed that there are much high nutritional constituents in the fermentation powder. Considering the influence of Chinese medicine inorganic elements, some relevant aspects of the fermentation powder may also have the efficacy, the wild mushroom fruiting body of I. hispidus contains higher protein value but the cadmium in it was exceeded.The fifth part of the test section, the toxicity and maximum dosage experiments were taken to evaluate the safety of the 2 kinds of fermentation powder of I. hispidus. The results showed that fermentation powder of I. hispidus does not make mice appear the phenomenon of poisoning or death, and the maximum daily dosage of sorghum and corn fermentation powder were up to 8g/Kg and 12g/Kg.The sixth part of the test section, the sorghum fermentation powder of I. hispidus were product development, and research on the production process of fermentation powder of I. hispidus of cookies. The single factor screening test and orthogonal test was taken, and obtained the best formula of cookies, which was the ratio of low gluten flour and the fermentation powder of I. hispidus was 100:10, the additive amount of butter was 80 g, the additive amount of sugar beam was 5g, the additive amount of whipping cream was 50 ml can make the quality to achieve best of the cookies.There were two sides to research on I. hispidus. On the one hand, by researching on the biological characteristics, and then cultivating the I. hispidus fruiting body successfully, it can lay the foundation of the sustainable utilization and mass production of I. hispidus; on the other hand, to find the reasonable alternatives of I. hispidus, and produce the fermentation powder, the research on the pharmacological activity of it found that the sorghum fermentation powder had better antitumor activity. Cookies were produced by the powder, which can find a way in food market, and also improve the utilization rate of this fungi.