贝莱斯芽胞杆菌Jt84高产伊枯草菌素的发酵优化  

Optimization of Fermentation for Iturin High Production of Bacillus velezensis Jt84

作　　者：张荣胜[1] 黄如宇 乔俊卿[1] 于俊杰[1] 齐中强[1] 杜艳[1] 俞咪娜[1] 宋天巧[1] 曹慧娟 潘夏艳 刘邮洲[1] 刘永锋[1] ZHANG Rongsheng;HUANG Ruyu;QIAO Junqing;YU Junjie;QI Zhongqiang;DU Yan;YU Mina;SONG Tianqiao;CAO Huijuan;PAN Xiayan;LIU Youzhou;LIU Yongfeng(Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]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》2025年第1期80-90,共11页Chinese Journal of Biological Control

基　　金：国家重点研发计划(2023YFD1400200);江苏省自主创新资金项目[CX(21)3083]。

摘　　要：贝莱斯芽胞杆菌Jt84对水稻稻曲病和稻瘟病具有良好防治效果,前期研究显示其产生的伊枯草菌素(iturin)是抑制稻曲病菌和稻瘟病菌生长和孢子萌发的主要活性物质。为了进一步提高抗真菌物质iturin含量,本文在贝莱斯芽胞杆菌Jt84发酵初期,通过外源添加合成iturin所需不同种类的前体氨基酸,利用Plackett-Burman单因素试验设计筛选获得3个影响iturin产量的主效前体氨基酸,分别为谷氨酸(glutamicacid,Glu)、丝氨酸(serine,Ser)和谷氨酰胺(glutamine,Gln)。进一步对主效前体氨基酸进行响应曲面优化,结果显示最优主效前体氨基酸添加浓度分别为Glu 0.08 g/L、Gln 0.17 g/L、Ser 0.19 g/L;其他氨基酸浓度为天冬酰胺(asparagine,Asn)0.06 g/L、脯氨酸(proline,Pro)0.06 g/L、酪氨酸(tyrosine,Tyr)0.006g/L。在此条件下,优化后发酵液的菌体含量为3.83×10^(9) cfu/mL,iturin产量为0.242g/L,iturin产量较优化前提高了26.04%;平板抑菌试验显示无菌滤液对稻曲病菌抑制带宽较初始培养基提高了23.66%。Bacillus velezensis strain Jt84 demonstrates remarkable control efficacy against rice false smut and blasts.Previous studies have established that iturin produced by Jt84 strain is the primary bioactive compound inhibiting rice fungal pathogens'growth and spore germination.To further augment the production of the antifungal agent iturin,this study investigated the early fermentation phase of B.velezensis Jt84 by supplementing exogenous precursors,precisely various amino acids required for iturin biosynthesis.Utilizing a Plackett-Burman single-factor experimental design,we identified three principal precursor amino acids that significantly influence iturin yield:glutamic acid(Glu),serine(Ser),and glutamine(Gln).Subsequent optimization via response surface methodology revealed the optimal concentrations for these essential precursors to be Glu at 0.08 g/L,Gln at 0.17 g/L,and Ser at 0.19 g/L,with additional amino acids present at concentrations of asparagine(Asn)at 0.06 g/L,proline(Pro)at 0.06 g/L,and tyrosine(Tyr)at 0.006 g/L.Under these optimized conditions,the fermentation broth achieved a bacterial content of 3.83×10^(9)cfu/mL and the iturin yield was 0.242 g/L,representing a 26.04%increase in iturin production compared to pre-optimization levels.Furthermore,the plate inhibition assay indicated that the inhibition width against Ustilaginoidea virens increased by 23.66%compared to the initial culture medium.

关 键 词：贝莱斯芽胞杆菌 伊枯草菌素 发酵 响应曲面法 

分 类 号：S476[农业科学—农业昆虫与害虫防治]