利巴韦林发酵工艺优化及菌株构建-生物工程专业毕业论文.docxVIP

ABSTRACTRibavirin ABSTRACT Ribavirin is an analog of purine nucleoside which plays important roles in the treatment of various kinds of DNA and RNA viruses．Where 1,2，4一Triazole一3-carboxamide (TCA)are added to the culture of nucleoside producing strmns of purine nucleoside，and ribavirin is synthesized from endogenous nucleoside catalyzed by the natural purine nucleoside phosphorylase(PNPase，encoded by pupG)of the strain．This fermentative method is named fermentation synthesis of ribavirin． The study firstly optimizate of fermentation condition for production ribavirin by our 1ab stock strain．Bacillus amyloliquefaciens TA208．LM iS the mutative strain of auxotroph of adenine．The adenine concentration in the fermentation broth iS critical as such：too little adenine affects the growth of bacteria,while too much restrains the synthesis of key enzymes．Firstly we optimized the type of adenine donor and its appropriate content．The results indicated that Springer 2506 was the best adenine donor，and 1 5 g／L was optimal for ribavirin accumulation．However,at the end of fermentation the adenine concentration released from this dose of Springer 2506 was too high．Therefore，the initial quantity of Springer 2506 Was reduced and extra adenine was added，the results indicated that the optimal initial adenine concentration Was 47．95 mg／L and accordingly the adenine in the fermentation liquid maintained 40-50 mg／L，which produced the highest PI冲P amidotransferase activity andthe ribavirin titer．This data indicated that controlling the initial addition of yeast extract and further feeding appropriate quantity of yeast extract Was likely a feasible way to control adenine concentration in the fermentation．We studied Springer 2506 fed．batch fermentation in 7．5 L fermentation tank．The results indicated that when 7．5 g／L Springer 2506 was used as substrate and another 7．5 egL fed，the ribavirin production increased by 1 1．29％compared to that when 1 5 g／L adding was added at a time． Plasmid pBSA43