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The Camptotheca acuminata endophytic fungi and their secondary metabolites

Author: LiXia
Tutor: LiuJiaJia
School: Central South University
Course: Pharmaceutical Engineering
Keywords: Camptothecin Endophytic fungi Response Surface Methodology Medium SRAP
CLC: R284
Type: Master's thesis
Year: 2011
Downloads: 60
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Abstract


Camptothecin as a specific DNA topoisomerase inhibitor is a good anticancer effect an active ingredient. However , due to its low content in the plant , Camptotheca acuminata , difficult to extract , and the structure is complicated , difficult to by chemical synthesis methods . Therefore , endophytic fungi as the camptothecin new medicinal resources has caused widespread concern. In this paper, about 50 endophytic fungi were isolated from Camptotheca acuminata bark . Camptothecin standard for the control , the use of thin-layer chromatography , UV spectrophotometry and high performance liquid chromatography , screened a camptothecin produced within the Health strain EFC I -13 . By morphological identification , classified as Penicillium spp . In order to improve the production of camptothecin , the fermentation medium of the strain EFC I-13 were optimized. By single factor experiment selected the best carbon source for glucose , a nitrogen source for the yeast extract and ammonium sulfate . Plackett-Burman experiments show that the main factors affecting the camptothecin for glucose , potassium dihydrogen phosphate and L- tryptophan . The steepest ascent experiment to determine the zero value of the main factors . Box-Behnken experimental and response surface analysis of optimum fermentation medium glucose 10.48 g / L , potassium dihydrogen phosphate 2.91g / L, L - tryptophan, 0.148 g / L, yeast extract 15g / L , magnesium sulfate 0.5 g / L, ammonium sulfate 0.8 g / L, calcium chloride 0.1 g / L , the maximum yield of camptothecin in this condition and to 80.41μ / L . After experimental verification , under the conditions of the medium , camptothecin production amounted 80.39μg / L, 43.9% higher than the yield pre- optimized medium . Randomly selected 36 PCR-SRAP primer Camptotheca acuminata endophytic fungi screened out of the six pairs of good stability, a primer combinations amplified bands and more . 6 pairs of primer combinations amplified 425 bands and 365 polymorphic bands , polymorphic bands ratio of 85.9% . In the similarity coefficient of 0.54 , the strains can be divided into three categories. SRAP methods can be used to the Camptotheca acuminata endophytic fungi polymorphism analysis .

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