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从烟草根际土壤中分离到1株生防细菌侧孢短芽孢杆菌(Brevibacillus laterosporus)B8,为了将B8应用于植物病害生物防治,对菌株B8产生的抑菌物质粗提物理化性质进行研究,并采用薄层层析、Pharmadex LH-20柱层析和高效液相色谱对抑菌物质进行分离纯化。试验结果表明:侧孢短芽孢杆菌B8抑菌物质粗提物具有较好的热稳定性,可耐受100℃的高温;其p H值适应范围较宽,在p H值4~10时都有较好的活性,在中性条件下活性最强;对紫外光和日光灯照射不敏感,可长时间保存;侧孢短芽孢杆菌B8产生的抑菌物质为蛋白类,经SDS-PAGE电泳确定蛋白分子量约为11.2k Da;质谱分析结果表明纯化蛋白在NCBI蛋白数据库中没有明确的匹配结果,推测此抑菌蛋白可能是在NCBI蛋白数据库中没有登录的1种新物质。
One biocontrol bacterium Brevibacterium laterosporus B8 was isolated from the tobacco rhizosphere soil. In order to apply B8 to the biological control of plant diseases, the physical and chemical properties of the crude extracts of the bacteriostasis strain B8 were studied. The antibacterial substances were isolated and purified by TLC, Pharmadex LH-20 column chromatography and high performance liquid chromatography. The results showed that the crude extract of B.cerevisiae B8 had good thermal stability and could tolerate high temperature of 100 ℃. The p H value was adapted to a wide range of pH values from 4 to 10 Has better activity, and has the strongest activity under neutral conditions; it is insensitive to ultraviolet light and fluorescent lamp and can be stored for a long time; and the bacteriostatic substance produced by B8 of Bacillus pleurotus is protein, and determined by SDS-PAGE electrophoresis The molecular weight of the protein was about 11.2kDa. The results of mass spectrometry showed that there was no clear match between the purified protein and the NCBI protein database. This bacteriostatic protein may be a new substance which is not registered in the NCBI protein database.