Study On Fermentation Conditions Of Taxol Endophytic Fungi Small Tank

Taxol is a diterpenoid compounds. At present, it is recognized as one of the broad-spectrum activity, strong anti-cancer drugs. Now, the production of Taxol mainly depends on the extract from the yew bark. Taxus species grow very slow, and taxol in the tree bark of the content is very low, in recent years, the world’s population and cancer incidence were explosive growth, the demand for Taxol also significantly increased. To solve the shortage of resources and the Taxus Paclitaxel increasing demand contradictions, scientists have done many research to improve the production of paclitaxel, including chemical total synthesis, Taxus cell culture and microbial fermentation. Producing taxol by microbial fermentation has obvious advantages, so people pay more and more attention for its research and development.In this paper, to utilize microbial fermentation to improve Taxol production, by single-factor test, P-B and response surface analysis, we can optimize to batch fermentation conditions of Taxol-producing bacteria HDF1-3. The main research methods and results are as follows:1.To optimize the nitrogen sources of fermentation medium, apply single-factor experimental design methods, utilize urea, soybean cake hydrolyzate, corn steep liquor and peptone medium in place of S-7 nitrogen in the material, the experimental results showed that soybean hydrolysis can affect the largest production of Taxol, Taxol production could reach 513.21μg/L;2.Using Plackett-Burman experiment to design of experiments about fermentation medium of linoleic acid and other 16 kinds of additives, the effects of significant factors are VB6, amphotericin and fructose amphotericin;3.In the base of Plackett-Burman experiments conducted, we can make steepest climbing and Box-Behnken experimental design, by the response surface method for the major impact factor for evaluation and optimization, and the quadratic polynomial regression model effecting fermentation strains HDF1-3 Taxol,at last we can use software SAS9.1 significantly to the model test. Results showed that VB6, amphotericin and fructose content of 17.34mg/L、16.63mg/L and 69.85mg/L, available to a larger production of Taxol;4.We can use single-factor test to optimize the fermentation process parameters, the best optimized fermentation conditions as follows:0～8d temperature of 28℃, speed of 120r/min, pass oxygen of 3L/min; 9～14d temperature of 26℃, speed of 100r/min, pass oxygen of 2L/min, Taxol production could reach 558.14μg/L;5.In 5L fermentor identified initial carbon source sugar addition and added sugar test, optimized the conditions of fed-batch processes, the best experimental design to add the initial conditions for the concentration of sugar is 13.1g/L, 8d in the first fermentation of sugar added concentration is 10g/L, will obtain the largest production of paclitaxel is 581.43μg/L, increasing 28.3% in production than the pre-optimization.