| Actinidia arguta, also known as soft dates, monkey pear, cane melon, rattan jujube. Actinidia arguta belongs to Actinidia, Actinidiaceae, which has the high nutritional value is very rich in VC, starch, pectin, anthraquinone compounds. Its function is very diverse:it can regulate the immune function, and also has anti inflammation, anti-tumor and anti-aging function. Endophytic fungi have been used as a variety of especially rich microbial resources, often parasitic in the roots, stems, leaves and other tissues and organs of the plant, so the endophytic fungus is an important source of natural active substances. Secondly, it is very rich in biological activity substances, which can be made of pesticides and other drugs. It has been demonstrated that can produce anthraquinone plant endophytic fungi. Anthraquinone is an important functional factor, has a variety of biological activities. For example, common shikonin has antibacterial and hemostatic effects of juglone and Sabino quinone has anti-tumor effect, senna has diarrhea effect. In addition the anthraquinones also has obvious antioxidant, antiviral and antitumor effect. Because of its easy cultivation, easy control, fast growth characteristics of Actinidia arguta endophytic fungi and its fermentation method is simple, high yield, which can not only shorten the fermentation time, and saves land resources. Therefore, it is very important to study to determine the extraction methods of anthraquinones in culture and fermentation broth of endophytic fungi in Actinidia arguta and fermentation method and optimization, biological activity.In this study, the fermentation of Actinidia arguta endophytic fungi anthraquinones extraction methods. Namely:Actinidia arguta endophytic fungi separation, purification and identification of the anthraquinone content detection method, endophytic fungi fermentation condition optimization and optimal extraction parameters established, anthraquinones antioxidant activity in vitro. The main results are as follows:This study through morphological identification and molecular identification of isolated endophytic fungi identification of Actinidia arguta. The experimental material for the isolation of endophytic fungi strains Z2. The identification results showed that three strains were ascomycetes. Finally, the Z2 strain was selected as the experimental object, and the Z2 strains were purified to the non pollution, so as to carry out the next experiment content:study on the extraction method of the compounds in the fermentation broth.In this study, the methods of extraction of the substance of the compounds were verified by stability test, reproducibility test and precision test. The test results showed that magnesium acetate colorimetry has reproducibility, precision, good stability. For this experiment is feasible and can meet the needs of the subsequent experiments.In this study, the optimum carbon source, nitrogen source, fermentation temperature, pH value and rotational speed of the optimum fermentation medium were screened out by measuring the yield of mycelia, and the optimum culture medium and fermentation conditions were determined. The experimental results show that the optimum composition of fermentation medium:the carbon source was glucose, the mycelium volume can reach 1.35g/100mL; the nitrogen source was beef extract, the mycelium of which reached 1.35g/100mL; the optimal culture conditions:the best fermentation temperature is 28 degrees Celsius.the mycelium weight is 1.30g/100mL; the best medium pH value is 6, the the amount of mycelium was 1.16g/100mL; the optimal speed of culture shock as the box 140r/min, the mycelium weight is 1.23g/100mL. Design of fermentation conditions of five factors and three levels of orthogonal test according to the result of single factor test, the L9(3)5 orthogonal test, to optimize the best fermentation condition, results show that:all kinds of factors on the mycelial growth affects the amount of each are not identical, which nitrogen source on the mycelial growth affects the amount of maximum, followed by fermentation temperature. That fermentation culture media and components of the optimum conditions for sucrose, peptone, fermentation temperature of 30 degrees Celsius and pH value is 6, speed is 150r/min.In this study, the extraction process conditions for the extraction of the substance of the compounds were optimized by single factor test and orthogonal design test. According to the extraction of the final content of the substance of the. The single factor experiment results showed that fermentation days excellent level followed by 5 and 8 days,10 days; the level of solvent extraction and followed by chloroform, ethyl acetate and methanol; liquid Biju level followed by 20:1 and 40:1 50:1.. Extraction of anthraquinones from Actinidia arguta endophytic fungi fermentation liquid absorption value is higher in this experimental condition. L9(3)3 orthogonal experiment showed that the effect of each factor on the extraction rate of the extraction rate. The extraction rate of liquid to material anthraquinones was mostly affected. The second is the duration of the fermentation, the last is solvent. Ptimal combination is the best condition for the extraction of the 5 day of fermentation. The extraction solvent is butanol and liquid feed ratio is 50:1.. Determination method of measuring the content of anthraquinones in the test are:drawing standard curve of anthraquinone standard of rhein, established the qualitative and quantitative methods of anthraquinones determination of Actinidia arguta endophytic fungi fermentation broth, anthraquinone content was calculated according to the standard curve.In this study, the antioxidant activity of DPPH free radical scavenging activity, superoxide anion scavenging ability, scavenging ability and reducing ability were studied. Test results show that:Actinidia arguta students fungus fermentation liquid of anthraquinone class material has higher DPPH radical scavenging ability and scavenging rate up to 70.80%; anthraquinones in Actinidia arguta endophytic fungus fermentation liquid with strong superoxide anion scavenging capacity, up to 76.26%; alkyl scavenging capacity of alkyl scavenging ability to 73.52%; Determination of the reduction ability of five kinds of different concentration gradient of anthraquinone compounds, reductive ability highest is 1.50. Four experimental results show that the effects of anthraquinones have strong antioxidant activity. From the above experiment results, anthraquinone has good antioxidant activity. |
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