万古霉素高产菌株选育及发酵配方优化.docx
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1、万古霉素高产菌株选育及发酵配方优化王会会程启东戴梦赵建强任风芝葛鹃鹏张志江摘要:为了提高东方拟无枝酸菌产万古霉素的发酵单位,降低生产成本,采用紫外诱变、常压室温等离子体诱变对东方拟无枝酸菌出发菌株V19-02进行诱变处理,通过筛选链霉素抗性突变菌株,选育高产菌株;通过对发酵培养基中碳源和氮源进行筛选,选取葡萄糖、淀粉、低温豆粉、酵母粉4个因素进行均匀设计。结果表明,获得的高产菌株V19-1-07摇瓶发酵单位提高33. 7%,且具有良好的传代稳定性;优化后的培养基配比质量分数为葡萄糖8. 0%.淀粉1.0%、低温豆粉L0%、酵母粉4.0%、氯化钠1.0%,磷酸二氢钾0.1%;小试结果证明,优化配
2、方可使50L罐发酵单位平均达到14110 gmL,比原始配方提高了 28. 9%o因此,研究获得的高产菌株V19-1-07可应用于万古霉素工业化发酵生产,以帮助企业降低成本,进一步提高竞争力。关键词:发酵工程;万古霉素;复合诱变;菌种选育;均匀设计;东方拟无枝酸菌中图分类号:TQ465. 9文献标识码:ADOI: 10. 7535/hbgykj.2022yx03011BreedingofhighvancomycinproducingstrainsandoptimizationoffermentationformulaWANGHuihuil, 2, 3, CHENGQidong4, DAIMen
3、gl, 2, 3,ZHA0Jianqiang4, RENFengzhil, 2, 3,GEKunpengl, 2, 3, ZHANGZhijiang4( 1. NewDrugResearchDevelopmentCompany , NCPC ,Shijiazhuang,Hebei052165,China;2. NationalEngineeringResearchCenterofMicrobiaIMedicine , Shijiazhuang ,Hebei052165,China;3. Hebei IndustryMicrobialMetabolicEngineeringTechnologyR
4、esearchCenter , Shijiazhuang, Hebei052165 ,China; 4. HuashengCompany , NCPC , Shijiazhuang ,Hebei052165, China)Abstract:InordertoincreasetheyieldofvancomycinbyAmycolatopsisorientalisandreducethecost,theoriginalstrainAycolatopsisorientalisV19-02wastreatedbyUVandatmosphericandroomteniperatureplasnias(
5、ARTP ) . Highyieldstrainswereselectedbyscreeningstreptomycinresistancetreatment. Meanwhile,thecarbonsourcesandnitrogensourcesoffermentationmediumwereoptimizedandfourfactorsincludingglucose,starch,low-temperaturesoybeanpowderandyeastpowderwereselectedforuniformdesignexperiment. Theresultsshowthatthey
6、ieldofvancomycinfermentationofthehigh-yieldmutantstrainAmycolatopsisorientalisV19-l-07isincreasedby33. 7%inflaskculture. Inaddition ,thestrainhasanexcellentpropagatingstability. Theoptimalfermentationmediumisasfollows : glucose8.0% ,starchi. 0% , low-temperaturesoybeanpowderl. 0% ,yeastpowder4.0% ,
7、NaCll. 0% ,KH2P040. 1%. Theexperimentalresultsshowthattheproductionofvancomycinreachsl4110 g/mLundertheoptimalconditionsbythestrainV19-l-07in50Lfermentor ,whichis28. 9%higherthantheoriginalformula. Afterappliedtolarge-scaleproduction ,itisconductivetoreducethecostandimprovethecompetitivenessofenterp
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- 万古霉素 高产 菌株 选育 发酵 配方 优化