等离子体作用结合氧限制模型选育利福霉素SV高产菌株  

Breeding of High-yield Rifamycin SV Strain by Plasma Action Combined with Oxygen Restriction Model

作　　者：栗波 王泽建[3] 梁剑光 刘爱军 李海东 LI Bo;WANG Ze-jian;LIANG Jian-guang;LIU Ai-jun;LI Hai-dong(School of Biological and Food Engineering,Changshu Institute of Technology,Changshu 215500,China;College of Pharmaceutical Science,Soochow University,Suzhou 215123,China;State Key Laboratory of Bioreactor Engineering,East China University of Science and Technology,Shanghai 200237,China;College of Pharmaceutical and Life Sciences,Changzhou University,Changzhou 213164,China;Hebei Xingang Pharmaceutical Co.,Ltd,Hebei 051530,China)

机构地区：[1]常熟理工学院生物与食品工程学院,常熟215500 [2]苏州大学医学部药学院,苏州215500 [3]华东理工大学国家生物反应器工程重点实验室,上海200237 [4]常州大学制药与生命科学学院,常州213164 [5]河北欣港药业有限公司,石家庄051530

出　　处：《中国生物工程杂志》2021年第2期38-44,共7页China Biotechnology

基　　金：国家重点研发计划(2017ZX07402003)资助项目。

摘　　要：利福霉素SV毒性低、疗效高、抗菌谱广,主要由地中海拟无枝酸菌发酵生产,其发酵过程属于耗氧发酵,供氧直接影响产物形成。为减少发酵过程氧限制影响,进一步提高利福霉素发酵产量,通过构建定向氧限制模型,将常温常压等离子体诱变和无水亚硫酸钠氧限制筛选模型相结合,建立了利福霉素生产菌株24孔板快速培养的高通量筛选方法,高效选育出能够耐受低氧环境的利福霉素SV高产菌株NSMXG-M126,发酵代谢状态参数变化显示,该高产菌株具有更好的氧亲和力。同样的供氧条件下,与对照相比表现出较快的菌体生长速率和利福霉素SV的快速合成能力。在低供氧情况下发酵单位达到7839mg/L,较出发菌株提高48%,表明耐受低氧的突变菌株具有更高的利福霉素SV生产效率。Rifamycin SV has low toxicity,high curative effect,and broad antibacterial spectrum.It is mainly produced through the aerobic fermentation by Amycobacterium marinum,and oxygen supply limitation was always the critical factor that affects the rifamycin SV biosynthesis.In order to reduce the influence of oxygen limitation in the fermentation process and further increase the fermentation yield of rifamycin,an oxygen limitation directional model was established with anhydrous sodium sulfite,combining normal temperature and atmospheric pressure plasma mutagenesis(ARTP);a high-throughput screening method of rapid culture of the vinegarproducing strains in 24-well plates was proposed;the high-yielding rifamycin SV strain NSMXG-M126 that can tolerate hypoxia has been efficiently selected.The changes in the parameters of the fermentation metabolic state show that the high-yielding strain has better oxygen affinity.Under the same oxygen supply conditions,it showed a faster bacterial growth rate and rapid synthesis of rifamycin SV compared with the control.In the case of low oxygen supply,the fermentation unit reached 7839 mg/m L,which was 48%higher than the original strain.The results demonstrated that the mutant strains that tolerate hypoxia have higher production efficiency of rifamycin SV.

关 键 词：利福霉素SV 地中海拟无枝酸菌 高通量筛选 ARTP 氧限制模型 

分 类 号：Q815[生物学—生物工程]