ARTP-DES复合诱变结合前体耐受性选育达托霉素高产菌株  

Selective breeding of high daptomycin-producing strain by ARTP-DES complex mutagenesis combined with precursor tolerance

作　　者：褚帅北 胡文婷 惠丰立[1,2] CHU Shuaibei;HU Wenting;HUI Fengli(College of Life Science and Agricultural Engineering,Nanyang Normal University,Nanyang 473061,Henan,China;Research Center of Henan Provincial Agricultural Biomass Resource Engineering and Technology,Nanyang Normal University,Nanyang 473061,Henan,China)

机构地区：[1]南阳师范学院生命科学与农业工程学院,河南南阳473061 [2]南阳师范学院农业生物质资源化河南省高校工程技术中心,河南南阳473061

出　　处：《精细化工》2023年第9期1969-1975,2002,共8页Fine Chemicals

基　　金：国家自然科学基金(31570021)。

摘　　要：为提高玫瑰孢链霉菌AN-126的达托霉素产量,对菌株AN-126进行常压室温等离子体-硫酸二乙酯(ARTP-DES)复合诱变,并结合癸酸钠耐受性筛选,最终获得一株达托霉素高产菌株RR-447,摇瓶产量为(101.41±0.37)mg/L,是出发菌株的2.44倍。对高产菌株RR-447进行平板传代、菌落形态及BOX-PCR多态性分析。结果表明,高产菌株RR-447高产性能稳定遗传,与出发菌株的菌球形态有明显差异,且高产菌株RR-447基因组DNA在1.50~2.00 kb之间存在一条特异性条带。在5 L发酵罐中通过分阶段补加癸酸钠溶液评价了菌株RR-447的发酵性能,达托霉素产量达到512.64mg/L。ARTP-DES复合诱变结合癸酸钠耐受性是选育达托霉素高产菌株的一种有效育种手段,为达托霉素工业微生物育种以及其他菌株改良提供了技术参考。In order to increase daptomycin production of Streptomyces roseosporus AN-126,a high daptomycin producing strain RR-447 was obtained from the complex mutagenesis of strain AN-126 induced by atmospheric and room temperature plasma-diethyl sulfate(ARTP-DES)and sodium caprate tolerance screening.The mutant strain RR-447 with a yield of(101.41±0.37)mg/L was obtained,which was 2.44 times that of the strain AN-126.The high yield strain RR-447 obtained was then subjected to plate passage,colony morphology and BOX-PCR polymorphism analysis.The results showed that the mutant strain RR-447,with a morphology different from the original strain,showed high yield and stable heredity.And there was a specific band found between 1.50 kb and 2.00 kb in the genomic DNA of RR-447 strain.The fermentation performance of strain RR-447 was evaluated by adding sodium caprate solution in batches in a 5 L fermentation tank,and the yield of daptomycin reached 512.64 mg/L.ARTP-DES complex mutagenesis combined with sodium caprate tolerance was an effective breeding method for high-yield daptomycin strains,which provided a technical reference for industrial microbial breeding of daptomycin and other strain improvement.

关 键 词：达托霉素 常压室温等离子体-硫酸二乙酯复合诱变 菌株选育 发酵 癸酸钠 生物工程 

分 类 号：TQ927[轻工技术与工程—发酵工程] Q815[生物学—生物工程]