多杀菌素生产菌的复合诱变选育及发酵优化
Breeding of Spinosad Producing Strains by Combined Mutation and Optimization of Fermentation
陈润杰 1李云飞 2张妍 2张善飞 3孙付保1
作者信息
- 1. 江南大学生物工程学院,江苏无锡 214122
- 2. 山东胜利生物工程有限公司,山东济宁 272000
- 3. 江南大学生物工程学院,江苏无锡 214122;山东胜利生物工程有限公司,山东济宁 272000
- 折叠
摘要
生物农药多杀菌素的发酵水平限制了其产业化发展.为了提高多杀菌素产量,以刺糖多孢菌 T1(Saccharopolyspora spinose T1)为出发菌,通过常压室温等离子体(atmospheric and room temperature plasma,ARTP)诱变和硫酸二乙酯(diethyl sulfate,DES)诱变,并以链霉素和鼠李糖作为筛选因子结合微孔板培养进行筛选,获得高产菌株S.spinosa F5;发酵后其多杀菌素产量为572.3 mg/L,比出发菌提高35.9％;通过单因素实验和响应面实验确定柠檬酸钠、异亮氨酸的最适添加量分别为0.46、0.10 g/L,豆油的最适添加体积分数为1.77％,此时多杀菌素产量为707.2 mg/L.在5 L发酵罐中进行分批发酵,发酵240 h产量接近800 mg/L;进一步分批补料发酵264 h,多杀菌素产量达1 175.5 mg/L.结果表明,ARTP和DES复合诱变育种及添加外源诱导物可以显著提高多杀菌素产量.
Abstract
The fermentation yield of spinosad,a biological pesticide,limits its industrialization development.To enhance spinosad production,Saccharopolyspora spinose T1 was mutagenized using atmospheric and room temperature plasma(ARTP)and diethyl sulfate(DES).The high-yield strain,S.spinosa F5,was obtained by streptomycin and rhamnose as screening factors combined with microplate culture.After fermentation,the yield of spinosad was 572.3 mg/L,an increase of 35.9％compared to that of the parent strain.Through single factor experiment and response surface test,the optimum addition amounts of sodium citrate and isoleucine were determined to be 0.46 g/L and 0.10 g/L,while the optimum addition volume fraction of soybean oil was 1.77％.Under this condition,the spinosad production reached 707.2 mg/L,and batch fermentation in a 5 L fermenter showed a yield close to 800 mg/L after 240 h fermentation.Further batch-fed fermentation resulted in a production of 1 175.5 mg/L after 264 h fermentation.The results showed that the compound mutation combining ARTP and DES breeding,along with the addition of exogenous inducers,could significantly increase the yield of spinosad.
关键词
多杀菌素/刺糖多孢菌/复合诱变育种/抗性筛选/培养基优化Key words
spinosad/Saccharopolyspora spinose/compound mutation breeding/resistance screening/medium optimization引用本文复制引用
出版年
2024
NETL
NSTL
万方数据