Page 121 - 《精细化工》2023年第9期

第 40 卷第 9 期 精 细 化 工 Vol.40, No.9
20 23 年 9 月 FINE CHEMICALS Sept. 2023

生物工程
ARTP-DES 复合诱变结合前体耐受性

选育达托霉素高产菌株

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褚帅北 ，胡文婷 ，惠丰立 1,2*
（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）

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 512.64 mg/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。
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
中图分类号：TQ927；Q815 文献标识码：A 文章编号：1003-5214 (2023) 09-1969-07

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

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CHU Shuaibei , HU Wenting , HUI Fengli 1,2*
（1. College of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang 473061, Henan,
China; 2. Research Center of Henan Provincial Agricultural Biomass Resource Engineering and Technology, Nanyang
Normal University, Nanyang 473061, Henan, China）

Abstract: 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.
Key words: daptomycin; atmospheric and room temperature plasma-diethyl sulfate complex mutagenesis;
strain breeding; fermentation; sodium caprate; bioengineering

收稿日期：2023-02-06; 定用日期：2023-04-03; DOI: 10.13550/j.jxhg.20230068
基金项目：国家自然科学基金（31570021）
作者简介：褚帅北（1998—），男，硕士生，E-mail：121975810@qq.com。联系人：惠丰立（1965—），男，教授，E-mail：huifl@
126.com。