芽胞杆菌工程菌株B9BD产表面活性素发酵培养基优化  

Optimization of Fermentation Medium for the Production of Surfactin by Engineered Strain B9BD of Bacillus subtilis

作　　者：乔俊卿[1] 孙凯 刘永锋[1] 陈夕军[2] 刘邮洲[1] QIAO Junqing;SUN Kai;LIU Yongfeng;CHEN Xijun;LIU Youzhou(Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;School of Horticulture and Plant Protection,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]江苏省农业科学院植物保护研究所,南京210014 [2]扬州大学园艺与植物保护学院,扬州225009

出　　处：《中国生物防治学报》2022年第2期383-392,共10页Chinese Journal of Biological Control

基　　金：江苏省自然科学基金(BK20201239);江苏省自主创新项目(CX(20)3126);国家重点研发项目(2017YFD0200400)。

摘　　要：生物农药“纹曲宁”的主要活性成分—枯草芽胞杆菌Bacillus subtilis Bs916可产生多种脂肽类抗生素。前期研究发现,degU和bmyA基因的突变株B9ΔdegUΔbmyA(B9BD)可以高效合成表面活性素(Surfactin)。本研究通过对培养基主要成分的单因素筛选并利用Design expert软件优化了工程菌株B9BD高效发酵产生Surfactin的培养基组分。通过Plackett-Burman试验、中心组合试验(CCD)和响应曲面法(RSM)优化了发酵Surfactin的最优培养基配方:葡萄糖24.27 g/L,黄豆粉29.05 g/L,牛肉浸膏3 g/L,NH_(4)NO_(3)4 g/L,L-亮氨酸2.36 g/L,KH_(2)PO_(4)3 g/L,NaCl 3 g/L,FeSO_(4)0.02 g/L。在优化培养基中,B9BD产生Surfactin的量高达到(934.25±32.67)mg/L,是其在LB培养基产量的5.6倍。以上研究结果将为Surfactin的高效发酵和应用提供理论基础和技术支撑。Bacillus subtilis Bs916 is the main component of the biopesticide“Wenquning”,which can produce a variety of lipopeptide antibiotics,such as Surfactin,Fengycin and Iturin.In a previous study,an engineering strain B9ΔdegUΔbmyA(B9BD)was found to be highly efficient in the synthesis of Surfactin.In this study,Plackett-Burman experiment,central xomposite design and response surface methodology were used to optimize the fermentation medium components of strain B9BD for Surfactin production.The optimized fermentation medium for high-yield Surfactin of strain B9BD was:glucose 24.27 g/L,soybean meal 29.05 g/L,beef extract 3 g/L,NH_(4)NO_(3)4 g/L,L-Leucine 2.36 g/L,KH_(2)PO_(4)3 g/L,NaCl 3 g/L and FeSO_(4)0.02 g/L.After optimization,the highest production of Surfactin reached(934.25±32.67)mg/L,which was 5.6 times that of the LB medium.The above research results will provide the theoretical basis and technical support for the efficient fermentation and application of Surfactin.

关 键 词：枯草芽胞杆菌 表面活性素 Plackett-Burman试验 中心组合试验 响应曲面法 

分 类 号：S476[农业科学—农业昆虫与害虫防治]