贝莱斯芽胞杆菌YL2021高产嗜铁素的摇瓶发酵工艺优化  

Optimization of Shake-flask Fermentation Condition for Siderophore Production of Bacillus velezensis YL2021

作　　者：沈佳慧 乔俊卿[2] 左杨 刘永锋[2] 刘邮洲[1,2] SHEN Jiahui;QIAO Junqing;ZUO Yang;LIU Yongfeng;LIU Youzhou(College of Plant Protection,Nanjing Agricultural University,Nanjing 210095,China;Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]南京农业大学植物保护学院,南京210095 [2]江苏省农业科学院植物保护研究所,南京210014

出　　处：《中国生物防治学报》2023年第4期861-874,共14页Chinese Journal of Biological Control

基　　金：国家自然科学基金项目(32272624);江苏省农业科技自主创新资金项目(CX(21)3096)。

摘　　要：贝莱斯芽胞杆菌Bacillus velezensis YL2021是一株对多种植物病原菌具有良好防治效果的生防菌,其基因组中含有完整的儿茶酚型嗜铁素合成基因簇,在缺铁条件下能产生具有抑菌作用的嗜铁素。为了提高贝莱斯芽胞杆菌YL2021嗜铁素的产量,采用摇瓶培养发酵,通过Plackett-Burman(PB)试验、中心组合试验(central composite design,CCD)和响应曲面法(response surface methodology,RSM)分析,优化菌株YL2021高产嗜铁素的发酵培养基成分和培养条件。结果表明,菌株YL2021高产嗜铁素的最优培养基配方:葡萄糖19.43 g/L、NH_(4)Cl 1.93 g/L、KH_(2)PO_(4)9.02 g/L、谷氨酸钠6.00 g/L、Mg SO_(4)1.75 g/L和Ca Cl20.08 g/L;最优培养条件为:温度30℃、初始p H值7.0、接种量5%、培养时间36.5 h、转速200 r/min和装液量75 m L(250 m L三角瓶中)。菌株YL2021在优化条件下嗜铁素产量达到734.86μmol/L,较优化前提高了10.34倍,优化效果明显。本文结果表明,已成功建立了适用于YL2021高产嗜铁素的摇瓶发酵体系,为嗜铁素的生防应用提供理论基础和技术支撑。Bacillus velezensis YL2021 has extensive antimicrobial activities against phytopathogens,and its genome harbors a catechol-type siderophore biosynthesis gene cluster.Strain YL2021 can produce siderophore in low-iron M9 medium,which demonstrated a wide range of antimicrobial activity in vitro.Using M9 medium as the basal fermentation medium,Plackett-Burman experiment,central composite design and response surface methodology were used to optimize the medium composition and culture conditions of strain YL2021 for high yield of siderophore in this study.The optimal medium composition of strain YL2021 for siderophore production was obtained as follows(g/L):glucose 19.43,NH_(4)Cl 1.93,KH_(2)PO_(4)9.02,sodium glutamate 6.00,MgSO_(4)1.75 and CaCl20.08.The optimal culture condition was as follows:culture temperature 30.0℃,initial pH value 7.0,culture time 36.5 h,rotation rate 200 r/min,75 mL operational volume in 250 mL shake flask with inoculum 5%.Based on the fermentation condition as described above,the value of siderophore production reached 734.86μmol/mL.Compared with that before optimization,siderophore production increased by 10.34 times.Together,our studies established an effective fermentation condition of strain YL2021 for siderophore production,which provide the theoretical basis and technical support for the efficient fermentation and application of siderophore in the future.

关 键 词：贝莱斯芽胞杆菌 嗜铁素 发酵优化 抑菌作用 

分 类 号：S476[农业科学—农业昆虫与害虫防治]