Mutation Breeding And Study On Optimize Fermentation Conditions For Strains Producing Podophyllotoxin

Podophyllotoxin is of great important in the field of medicine because of its antivirus and anti-cancer activities. It was derived mainly from the Podophyllotoxin plants. But the natural sources were limited. Endophytic fungi have the activity that can produce same or similar compounds to the plant. They are easy to be raised and controlled and their effective component output may be raised through mutation breeding. It is a new method by fermentation of endophytic fungui. So far,many research focused on separating valuable endophytic fungi which can produce Podophyllotoxin. Through compound mutation to producing-srains,then sudy on the optimizational fermentation conditions of high production strains,in the hope of providing some theoretic reference on producing botanic activity compouds by microbiology fermentation.Objectives:Compound mutation to producing-srains,then sudy on the optimizational fermentation conditions of high production strains in order to provide a new method obtaining anti-cancer material——podophyllotoxin and the technical data obtained from research on producing botantic activity compounds by microbiology fermentation. This will be lay the foundation for industrialization. Methods:High producing strain is attained by UV and He-Ne laser .The components were identified by thin layer chromatographic(TLC),Thin layer chromatographic scanning,high performance liquid chromatographic(HPLC) that consist in fermentation of high producing strain;The thesis designed semi-synthesized medium compatibhe to the strain, and optimized composition respectively. Results:Ty4-16-26 was attained after UV mutation with its biomass enhancing 31% and production-biomass enhancing 25%. Laser mutation based on this carries obtained high production Ty4-16-26, with biomass enhancing 17% and production-biomass enhancing 20%. This theis confirm that the mutated strain can produce podophyllotoxin. The optimizational ingredient of the semi-synthesized medium were as follows: 2% soluble srarch, 2 %sucrose, 0.1% peptone, 0.1% yeast extract, 0.1% NH₄(SO4)₂, 0.03% K₂HPO₄, 0.03% Nacl , 0.03% MgSO₄, pH6.5. The biomass increase 20.3% and the production-biomass ratio increase 9.7% by using the optimizational medium at temperature of 28℃for 7d. The scale-up trial in 5L fermentor showed that mycelium grew well and products produced steadily. Conclusions:The method that obtain high-producing with mutation breeding;The technics that obtain podophyllotoxin with fermentation and the tenique for extraction were gained. This research has the potential of industrialization.