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Biosynthesis of Polyphenols of Inonotus Obliquus in Submerged Cultures and Biological Activity

Author: ZhuJinWei
Tutor: XuXiangQun
School: Zhejiang University of Technology
Course: Applied Chemistry
Keywords: Inonotus obliquus submerged culture polyphenols lignocellulose antioxidant activity
CLC: TQ920
Type: Master's thesis
Year: 2011
Downloads: 23
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Abstract


Inonotus obliquus is a rare medicinal and edible fungus. The fruit body and mycelium of Inonotus obliquus contain several kinds of bioactive substances. Polyphenols in the fungus have an exellent antioxidative activity, anti-inflammatory, antiviral activity, antitumor, platelet aggregation inhibitory and immune-stimulating effect. We study the synthesis of polyphenols with shaker and fermentor fermentation. The content of total extracellular polyphenols(EPP) and intracellular polyphenols(IPP) and antioxidant activity as an index in this progress.Depended on thses indexes, we can find and establish the best submerged culture system of Inonotus obliquus.We study the characteristics of submerged fermentation on the basic culture medium. In nature, white rot fungi can degrade the lignicellulose.In the progress, lignicellulose is degraded into small molecular compounds including glucose and other carbohydrates. All these compounds can provide the necessary follow-up carbon sources for the growth of white rot fungi.In this progress, the cotent and the antioxidant activity of EPP and IPP was enhanced. Moreover, the basic composition and content of extracellular phenolic compounds in the lignocellulose medium and the basal mediumhe are also been analyzed by the LC-MS approach.It gives the reasons of big difference in cotent and the antioxidant activity of EPP and IPP.The original strains of Inonotus obliquus (Inonotus obliquus CBS 314.39) were activated and cultivated. Mea plate bacterial colony and mycelial morphology were observed. And they were preserved with the fungi preserved method. We also study the basis of biomass, the cotent of EPP and IPP and other diverse parameters of fermentation to determine the fermentation period of 12 days.We investigate the effect of shaker and fermenter to the content of EPP and IPP.It showed that, fermenter fermentation can produce more EPP and IPP than shaker fermentation.Especially in the lignocellulose medium, fermenter fermentation produced more than 2.5 mg GAE/g and 16.8 mg GAE/L of IPP and EPP, respectively. We pay much more attention on the promoting effects of Inonotus obliquus degrade lignicellulose, which enhance the content and antioxidative activity of EPP and IPP. Experimental results show that the addition of lignicellulose increased the EPP and IPP remarkably. Compared to the basal medium, increased from 13.5 mg GAE/g to 23.7 mg GAE/g of EPP. And IPP increased from 42.5 mg GAE/L to 135.7 mg GAE/L. At the same time, in the experiment of scavenging hydroxyl radical with the EPP and IPP, the IC50 value of EPP with 216 h incubation in the lignicellulose medium is 169μg/mL much lower than the value 260μg/mL with 216 h incubation in the basal medium. The IC50 value of IPP with 288 h incubation in the lignicellulose medium is 199μg/mL lower than the value 233μg/mL with 72 h incubation in the basal medium.We compared with shaker and fermentor two submerged fermentation, two sources of liquid medium (basal medium, lignicellulose medium) and composition and content of EPP and IPP. The basic compontents of EPP are 17.4% EGCG,12.7% ECG,12.0% Inoscavin B,13.1% Phelligridin G and 7.9% Davallialactone in the lignicellulose medium compared to 10.6% EGCG,4.5% ECG,10.2% Phelligridin G, 17.2% Gallic acid in the basal medium; The basic compontents of IPP are8.7% Phelligridin G,23.8% Gallic acid,10.7% Kaempferol,15.3% p-Coumaric acid in the lignicellulose medium compared to compared to 4.3% Phelligridin G,38.2% Gallic acid,15.2% Ferulic acid,17.6% Rhoifolin in the basal medium.The results show that the polyphenols in lignicellulose medium is more complex than the basic medium. And the composition of EPP have more species than the IPP. Fermentor fermentation of polyphenols and shaker fermentation of polyphenols have more complicated in type, but the EPP and IPP of fermentor fermentation content have more content than shaker fermentation.

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