棉花黄萎病拮抗细菌B.velezensis6-61拮抗蛋白发酵条件的优化  

Fermentation Conditions Optimization of Antagonistic Bacteria B.velezensis 6-61 Against Verticillium dahliae Kleb

作　　者：李红亚[1] 李术娜[1] 朱宝成[1] 

机构地区：[1]河北农业大学生命科学学院,河北保定071001

出　　处：《中国农学通报》2011年第21期93-99,共7页Chinese Agricultural Science Bulletin

基　　金：河北省自然科学基金项目"棉花黄萎病拮抗细菌的筛选及抗菌蛋白的分离"(398152)

摘　　要：为了提高棉花黄萎病拮抗细菌B.velezensis6-61活性物质的产量,为进一步开展6-61菌株的后续研究奠定基础,对6-61菌株产生的活性物质的性质进行了分析,并对其发酵条件进行了优化研究。采用有机溶剂萃取、硫酸铵沉淀对菌株6-61发酵液进行处理并检测抑菌活性,初步确定了该拮抗物质为蛋白质。采用单因素试验对6-61菌株的发酵培养基所需的碳源、氮源、无机盐进行了筛选,并采用L16(44)正交实验对棉花黄萎病拮抗细菌6-61的发酵培养基组成以及发酵培养条件进行了优化。最终优化后的培养基组成为:麦芽糖3%、大豆蛋白胨3%、NaCl0.01%、KCl0.01%;最佳发酵条件:pH7.0、接种量8%、种龄16h、装瓶量30mL/250mL。在优化后的培养基及培养条件下进行发酵,拮抗细菌6-61的发酵液抑菌圈直径可达到36mm。与其他一些常见培养基的发酵效果相比较,在优化后的发酵条件下6-61菌株拮抗物质的产量明显高于其他常见培养基。In order to improve the yield of antagonistic substance produced by B.velezensis 6-61 which has antagonistic activity against Verticillium dahliae Kleb,the analysis of the properties of antagonistic substance and fermentation conditions optimization of stain 6-61 were studied to provide a basis for further study of the antagonistic mechanism.First,the fermented broth of strain 6-61 was treated by two following methods：extraction with organic solvent and precipitation by ammonium sulfate respectively.The results indicated that the component of strain 6-61 responsible for antagonistic activity was considered as protein.Then,the optimum fermentation medium ingredients,such as carbon sources,nitrogen sources and inorganic salts,were determined by using the single-factor methods.And the optimal medium compositions and other fermentation conditions of antagonistic bacteria 6-61were optimized through L 16（4 4） orthogonal experiment.The final optimization of the medium was obtained as follows：malt sugar 3%,soybean protein peptone 3%,NaCl 0.01%,KCl 0.01%,pH 7.0,inoculation volume 8%,seed culture time 16 h and bottle filling capacity 30 mL/250 mL.In the optimal medium compositions and culture conditions,the inhibition zone diameter of the fermentation liquid could reach 36mm.And the final fermentation medium has more fermentation productions compared with some other common media.

关 键 词：大丽轮枝菌 拮抗菌 发酵条件 优化 

分 类 号：TQ920.6[轻工技术与工程—发酵工程] S482.28[农业科学—农药学]