| Bi-directional fermentation is a new Chinese herbal fermentation technology, with a lower toxicity?expand the property of a medicine?improve absorption trait. The study had tested the dynamic changes of antioxidant activity and antibacterial activity of substance, and analyzed the correlations between different active ingredients and antioxidant activity?antibacterial activity in fermentation substance.It aimed proven mechanism of activity and antibacterial activity of the spirit return mycoplasma antioxidant, to provide a reference for the development and efficient use of resources in the spirit of traditional Chinese medicine products owned by mycoplasma.The important research results as following:1. The change rule of water and ethanol extract of ganoderma and angelica sinensis substance were determined systematacially. The results showed that, at the earlier fermentation(G0),the antioxidant activity of fermentation substance gradually increased, reached optimal at metaphase fermentation and later(G12-G16) decreased. Beside,it is showed that the antioxidant activity of water extract of fermentation substance significantly higher than ethanol extract.2. The antibacterial activity and stability of water and ethanol extract of ganoderma and angelica sinensis substance were evaluated systematacially. The results showed that, ethanol extract of ganoderma and angelica sinensis substance to three kinds bacterias,like Staphylococcus aureus?Escherichia coli?Bacillus subtilis,all possessed certain inhibition,and gradually increased as the extension of fermentation time. G16 of inhibition rate of three bacterias were 62.5%, 52.08%, 81.1%.The ethanol extract of ganoderma and angelica sinensis substance to the group ofthree kinds fungus such as Penicillium expansum?Alternaria alternate?Aspergillus niger and there germination rate of spores all had certain inhibition and rised as the fermentation time go. At the same time, research showed that fermentation fungal substance had good stability to heat, acid, alkali and ultraviolet radiation.3. The change rule of main active fractionsof ganoderma and angelica sinensis substancewere traced dynamically. At metaphase fermentation,The content of flavonoid?total phenols?total sugar and triterpenoids all increased with fermentation time prolonged. The content of total sugar reached maximum at middle time(G8).The content of flavonoid?total phenols and triterpenoids reached maximum at latter time,the triterpenoids reached optimal at G16.Had few ferulic acid content in matrix, and the ferulic acid content gradually reduced with the fermentation process.4. The correlations between the contention of main active ingredients and antioxidant activity?antibacterial activity in fermentation substance wered analyzed. The results showed that, The content of total sugarsignificantly correlated with DPPH scavenging rate and hydroxyl free radical scavenging. The content of total phenols significantly correlated with hydroxyl free radical scavenging and superoxide. The content of triterpenoids were significantly correlated with bacteriostasis for Alternaria alternate?Penicillium expansum?Bacillus subtilis?Staphylococcus aureus?Escherichia coli and inhibition of spore germination rate.5. In this part, a two-stage culture process by combining shake culture with static culture was proposed to study the effects of the four kinds of Chinese herbs(Angelica sinensis, Codonopsis pilosula,Glycyrrhiza uralensis and Radix Astragali) on production of ganoderma triterpenoids and mycelium growth. The results showed that the optimum two-stage cultivation conditions were fermentation broth C / N was 4: 1, two-stage shift time was the 3rd day,culture volume was 50 m L/250 m L.Under the conditions, four kinds of extracts could all significantly enhance cell growth biomass and the production of ganoderma triterpenoids, but their effective concentration ranges were different.The alcohol extracts of Angelica sinensis and Codonopsis pilosula were effective within 80g/L, however, the water extracts of Glycyrrhiza uralensis and Radix Astragali could enhance ganoderictriterpenoids production within 15g/L. The best adding time of Angelica sinensis extract was on the 2nd day, and the others were on the 3rd day.Verification test in 1.5 L fermentor showed that four herbs extracts are effective elicitors, which can promote the production of ganoderic triterpenoids, especially Angelica sinensis alcohol extracts was 80g/L in fermentation broth, the extracellular triterpenoids yield reached 965.69 mg / L.In conclusion, Ganoderma and Angelica sinensis substance was a new medicine with nutrient-rich and functional.Primary material was natural and unpolluted, ganoderma and angelica sinensis fungal substance had great antioxidant activity and antifungal activity, this was conform to contemporary consumers demand for food safety, nutrition and health. And this product is expected to develop into food preservatives and antioxidants, has the high value-added products and good market application prospect. |
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