Fermentation Condition Optimization And Biological Activity Components Analysis For Phellinus Lgniarius

Phellinus baumii, a mushroom grow on mulberry tree in orangecolor, was used in abdomnalgia and bioody gonorrhea in traditional Chinese medicine, Modem medical research indicated that it had curing effect against stomach aches, inflammation, tumors and so on. It is also used to improve overall health and prevent various diseases, such as lymphatic diseases, immunoregulation and Liver protection. Up to now, many researches were focused on the chemical composition, structure, pharmacology of polysaccharides and proteoglycan of fruiting bodies. Although p. baumii have a a great medicinal value, it has not been comprehensively investigated due to difficult culture and collection. Mycelial culture is a good alternative way to acquire sufficient amounts of P. baumii, to accelerate development and utilization.Therefore, there are three parts in this dissertation including optimization of submerged fermentation parameters, antibacterial and antioxidant activity of various extracts of P.baumii, and the chemical constituents and structure identification in the ethyl acetate extract of P.baumii.1. Submerged fermentation condition of P.baumii in flask culture was optimized. The optimized fermentation condition of producing mycelial growth, flavones and exo-polysaccharide was:corn powder5%, yeast extract0.4%, KH2P040.2%, MgSO4·H2O0.1%. The optimized conditions were temperature28℃, rotation speed150r/min, initial pH normal, inoculation volume10%, loading volume100mL/250mL, and fermentation time6days.2. Investigate possible antioxidant and antibacterial activity of various extracts of P.baumii. In this study, reducing power, lipid per-oxidation,2,2'-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid)(ABTS+)free radical scavenging assay,1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activities, hydroxyl radical scavenging were investigated. Extracts of P.baumii were also tested for antimicrobial activity by Disc Diffusion method on the following test microorganisms: Micrococcus luteus AS1.1848, Staphylococcus aurenu CMCC(B)26003, Lactobacillus brcvis AS1.12, Escherichia coli CICC21524, Pseudomonas aeuginosa ATCC9027, Serratia marcescens ATCC14041, Hansenula anomala AS2300. The results indicated that extracts had a good antioxidant activity. In the lipid per-oxidation, extracts of P.baumii all displayed high antioxidant activity with IC50all below0.16mg/mL. In the ABTS'+free radical scavenging assay, EtOAc fraction displayed the highest total antioxidant activity. In the DPPH free radical scavenging assay, reducing power and hydroxyl radical scavenging activities, EtOAc and n-BuOH fraction displayed higher antioxidant activity. The n-BuOH extract of P. baumii showed inhibition zone against Staphylococcus aureu CMCC(B)26003, Escherichia coll CICC21524and Hansenula anomala AS2.300. The MICs were0.3125mg/mL,0.6250ing/mL,0.3125mg/mL, respectively.3. Five chemical components were purified from ethyl acetate extract of P. bournii by HSCCC, Sephadex LH-20and Semi-Pre-HPLC as follows:inoscavin A, caffeic acid, naringenin, coumarin,3,4-dihydroxy benzaldehyde. Their structures were determined by UV, MS and NMR analysis, respectively.