春雷霉素高产菌株选育及其工业生产应用
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摘要
为了获得适用于工业化生产的春雷霉素高产菌株,从陕西渭南某生物农药厂发酵生产车间不同发酵罐批中分离筛选到5株春雷霉素生产性能优良的小金色链霉菌,以这些菌株为出发菌株,利用递归原生质体融合法,获得了3株遗传稳定的高产春雷霉素重组菌株,其中1株重组菌株K2018S F5-06的春雷霉素产量达到了19.6 g/L.经30吨罐生产验证,平均发酵效价比原始出发菌株K2010 S 17-182春雷霉素产量提高了37.1%.采用自然分离筛选结合递归原生质体融合育种技术能有效提高目标代谢产物的产量,产生巨大经济效益.
In order to obtain the high-yield strain suitable for industrial production of Kasugamycin,five strains of Streptomyces fuliginea with excellent performance for producing Kasugamycin were isolated from different fermenter batches in a fermentation company producing agricultural antibiotics in Shaanxi Province.Three genetically stable recombinant strains for high-yield of Kasugamycin were successfully selected by recursive protoplast fusion.The yield of one of the recombinant strains K2018 S F5-06 reached 19.6 g/L.After 30 tons of pot production verification,the average fermentation potency increased by 37.1% compared with the original starting strain K2010 S 17-182.It indicates that the natural separation and screening combined with recursive protoplast fusion technology can effectively increase the yield of target metabolites and obtain huge economic benefits.
In order to obtain the high-yield strain suitable for industrial production of Kasugamycin,five strains of Streptomyces fuliginea with excellent performance for producing Kasugamycin were isolated from different fermenter batches in a fermentation company producing agricultural antibiotics in Shaanxi Province.Three genetically stable recombinant strains for high-yield of Kasugamycin were successfully selected by recursive protoplast fusion.The yield of one of the recombinant strains K2018 S F5-06 reached 19.6 g/L.After 30 tons of pot production verification,the average fermentation potency increased by 37.1% compared with the original starting strain K2010 S 17-182.It indicates that the natural separation and screening combined with recursive protoplast fusion technology can effectively increase the yield of target metabolites and obtain huge economic benefits.
引文
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[11] Zeng W,Chen G,Wu H,et al.Improvement of bacillus subtilis for poly-γ-glutamic acid production by genome shuffling[J].Microbial Biotechnology,2016,9(6):824-833.
[2] 崔松华,刘丽萍.2%春雷霉素WP防治白菜软腐病药效试验[J].农药,2007,46(6):416-417.
[3] 袁超,袁丹丹,赵建刚,等.硫酸二乙酯与紫外线复合诱变选育糖化酶高产菌株的研究[J].食品与药品,2018,20(3):219-223.
[4] Zhang Y X ,Perry K ,Vinci V A ,et al.Genome shuffling leads to rapid phenotypic improvement in bacteria[J].Nature,2002,415(6 872):644-646.
[5] 龚国利,陈松,李慧,等.基因组重组技术选育埃博霉素B高产菌株[J].中国抗生素杂志,2013,38(2):106-110.
[6] 龚国利.粘细菌的Genome shuffling育种技术及其埃博霉素的高产改良[D].济南:山东大学,2007.
[7] Kentaro I,Ryo I,Takahiro B,et al.Improvement of xylose fermentation ability under heat and acid co-stress in saccharomyces cerevisiae using genome shuffling technique[J].Frontiers in Bioengineering and Biotechnology,2017,5:81.
[8] 公维亮,薛正莲,周扬,等.原生质体诱变及融合选育那西肽高产菌株[J].中国抗生素杂志,2016,41(5):335-339.
[9] Huang Q G,Zeng B D,Liang L,et al.Genome shuffling and high-throughput screening of brevibacterium flavum MDV1 for enhanced l-valine production[J].World Journal of Microbiology & Biotechnology,2018,34(8):121-124.
[10] 王敏,张若兰,崔志峰,等.基因组重排技术在酵母菌育种中的应用[J].工业微生物,2018,48(1):63-68.
[11] Zeng W,Chen G,Wu H,et al.Improvement of bacillus subtilis for poly-γ-glutamic acid production by genome shuffling[J].Microbial Biotechnology,2016,9(6):824-833.