Studies On Fermentation And Secondary Metabolites Of Endophytic Fungi Producting Vincamine Compounds

Vincamine and its derivatives have been played a great role in the treatment of cardiovascular,cerebrovascular disease and dementia. The demanded amount is large and the market of them are vastitute. Until now the methods of manufacturing of them are still rely on the extraction of Vinca minor and then semi-synthesis. Due to the limit of plant materials, production processes and costs have a great defect. In the experiment, specific fungal endopHytes producing Vincamine were screened from Vinca minor, followed by microbial fermentation technology to extract Vincamine and its compounds.This study is divided into the following sections:(1) The specific fungal endophytes producing Vincamine has been screened. Thin layer chromatography (TLC) and liquid chromatography-mass spectrometry (LC-MS) method were applied to preliminary qualitative analysis towards the secondary metabolites of several group fungal endophytes. By the method mentioned,the strain produce vincamine compounds was found.(2) the influence brought by several factors during fermentation, the target strains was discussed in the paper. Fermentation condition of the target strains were optimized by using response surface methodology for the best condition, and proved. The yield of target product was improved.(3) several method for the purification of secondary metabolites.TLC analysis showed that the metabolites of strain VINI-6and the control displacement products have similar values, which possibly suggest they were similar materials,LC-MS analysis showed that the target product peak time was2.21min, and the spectrum showed that substances with molecular mass367may exist in the peak time, which was consistent with ethyl Changchun amine, therefore we guess it may be Changchun amine compounds. On the basis of single factor experiment, we choosed fermentation time, fermentation temperature, pH of stable fermentation period for Box-Benhnken design of experiments with Design-Expert software for analysis. Finally we got the cultural condition where the Changchun amine compounds yield of VINI-6was highest: 26.2℃fermentation temperature, fermentation time7.56d, pH3.2after the stationary pHase. After the validation constant yield was about70.58μg (in the extraction of1.5L Mycelial Fermentation). After the purification method of fermented products in Apocynaceae plant endopHytic fungi VINI-6, we found that organic solvent extraction was the best extraction method.