A Camptotheca Endophyte Produce And Analysis Of Active Substance

This thesis focuses on the basis of the research camptotheca acuminate endophytesproperties characteristics, using microbial fermentation of camptotheca acuminata tenendophyte liquid culture carried out a study aimed at screening produced from CPT,camptothecin hydroxyl endophyte. Purpose is to search the new medicine source ofcamptothecin use the rich microbial resources, to provides the theory basis forHydroxycamptothecin and Camptotheca acuminata endophyte, producting by microbialfermentation, in industrial fermentation, and lay a certain foundation for the screening ofactive substances or new compounds with medicinal value.As a development great power, with the accelerating industrialization, urbanization andpopulation aging process, due to their own lifestyle and habits, as well as a series of problemssuch as environmental pollution, the number of cancer continues to grow, cancer has becomeone of the common death causes of residents in our country. In recent years, as a newcontinuously developed effective anti-cancer drugs, and combined with treatment technologydevelopment and improvement,drug therapy to become one of the considerable developmentpotential in numerous cancer treatment.Hydroxycamptothecin is alkaloids with a strong antitumor activity, isolated from theCamptotheca acuminate, a tree of plants in China, by the Wall and co-works in1966first. Inrecent years, although the chemical synthesis methods of camptothecin and its derivativesconstantly updating, most of them still exist a series of problems, such as higher difficulty,cycle length, more cost, low success rate. Therefore, we attempted study more efficient andpractical new method to obtoain alkaloid. We draw a conclusions from this test:（1）High-performance liquid chromatography’s test condition is that: mobile phase isacetonitrile:water (v/v)=70:30, C18column (4.6mm×150mm,5microns), the flow rate is1mL/min, detection wavelength of camptothecin is256nm, detection wavelength of Hydroxycamptothecin is256nm, every time’s Sample quantity is20μL. Due to this phase is onlyqualitative, so chosen to use reverse phase high performance liquid chromatography (HPLC)wavelength switch method, simultaneous determination of camptothecin and hydroxycamptothecin, so as to shorten the testing time，can improve the efficiency of detection. （2）In this article, determinated metabolites of ten endophytic liquid cuture withhigh-performance liquid chromatography, preliminary screening and determine showed thatseven endophytes extracts have a characteristics of camptothecin or hydroxyl camptothecin.Among them, the four produce camptothecin and hydroxyl camptothecin, the two strainsproduce hydroxyl camptothecin, and a one produce camptothecin only.（3）On this basis, replace liquid medium to the bacterial strain that can producecamptothecin and hydroxyl camptothecin, continuous to cultivate again, found thatcamptothecin content is the highest on the sixth day after shake cultivation, hydroxycamptothecin content is the highest on the twelfth day after shake cultivation, content ofhydroxy camptothecin and camptothecin in the fermented liquid higher than the mycelium.