万古霉素高产菌株选育及发酵配方优化  

Breeding of high vancomycin producing strains and optimization of fermentation formula

作　　者：王会会 程启东 戴梦[1,2,3] 赵建强 任风芝 葛鹍鹏[1,2,3] 张志江 WANG Huihui;CHENG Qidong;DAI Meng;ZHAO Jianqiang;REN Fengzhi;GE Kunpeng;ZHANG Zhijiang(New Drug Research & Development Company,NCPC,Shijiazhuang,Hebei 052165,China;National Engineering Research Center of Microbial Medicine,Shijiazhuang,Hebei 052165,China;Hebei Industry Microbial Metabolic Engineering & Technology Research Center,Shijiazhuang,Hebei 052165,China;Huasheng Company,NCPC,Shijiazhuang,Hebei 052165,China)

机构地区：[1]华北制药集团新药研究开发有限责任公司,河北石家庄052165 [2]微生物药物国家工程研究中心,河北石家庄052165 [3]河北省工业微生物代谢工程技术研究中心,河北石家庄052165 [4]华北制药华胜有限公司,河北石家庄052165

出　　处：《河北工业科技》2022年第3期253-260,共8页Hebei Journal of Industrial Science and Technology

基　　金：石家庄市微生物药物技术创新中心绩效后补助项目(217790377B)。

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1.0%、酵母粉4.0%、氯化钠1.0%、磷酸二氢钾0.1%;小试结果证明,优化配方可使50 L罐发酵单位平均达到14110μg/mL,比原始配方提高了28.9%。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。In order to increase the yield of vancomycin by Amycolatopsis orientalis and reduce the cost,the original strain Amycolatopsis orientalis V19-02 was treated by UV and atmospheric and room temperature plasmas(ARTP).High yield strains were selected by screening streptomycin resistance treatment.Meanwhile,the carbon sources and nitrogen sources of fermentation medium were optimized and four factors including glucose,starch,low-temperature soybean powder and yeast powder were selected for uniform design experiment.The results show that the yield of vancomycin fermentation of the high-yield mutant strain Amycolatopsis orientalis V19-1-07 is increased by 33.7%in flask culture.In addition,the strain has an excellent propagating stability.The optimal fermentation medium is as follows:glucose 8.0%,starch 1.0%,low-temperature soybean powder 1.0%,yeast powder 4.0%,NaCl 1.0%,KH_(2)PO_(4) 0.1%.The experimental results show that the production of vancomycin reachs 14110μg/mL under the optimal conditions by the strain V19-1-07 in 50 L fermentor,which is 28.9%higher than the original formula.After applied to large-scale production,it is conductive to reduce the cost and improve the competitiveness of enterprises further.

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 

分 类 号：TQ465.9[化学工程—制药化工]