侧孢短芽孢杆菌BL-21抗菌蛋白的稳定性分析和分离纯化

Stability Analysis, Isolation and Purification of Antimicrobial Protein From Brevibacillus laterosporus BL-21

为明确侧孢短芽孢杆菌BL-21(Brevibacillus laterosporus)产生的抗菌物质的类型,对其发酵液抗菌活性的稳定性进行分析,并在此基础上对有效成分进行分离纯化。采用杯碟法检测了B.laterosporus BL-21代谢产物的热稳定性、酸碱稳定性和蛋白酶稳定性;通过硫酸铵盐析、CM-Sepharose Fast Flow离子交换层析、凝胶过滤层析和SDS-PAGE进行了分离纯化。研究结果表明:B.laterosporus BL-21产生的抗菌物质在50℃以下稳定,在pH 7~8时抑菌效果最好,对蛋白酶敏感;发酵液中含有多组分抗菌物质,其中一个组分的分子量为20.1 k Da。B.laterosporus BL-21的发酵液能抑制烟草赤星病菌(Alternaria solani)和蜡样芽孢杆菌(Bacillus cereus)的生长,具有一定的生防潜力和研究价值。

To clarify the type of antimicrobial substance produced by Brevibacillus laterosporus BL-21, the stability of the fermentation liquor was analyzed, and the active i this. The thermostability, pH stability and protease stability of ngredients were isolated and purified based on antimicrobial metabolites from B laterosporus BL-21 were detected by cylinder plate method. The antimicrobial substances were isolated and purified by ammonium sulfate precipitation, CM-Sepharose Fast Flow ion exchange chromatography, gel filtration chromatography and SDS-PAGE. The results showed that the antimicrobial substance produced by B laterosporus BL-21 was stable below 50~C, its an protease. Several antimicrobial components were tifungal activity was the best at pH 7-8, and it was sensitive to isolated from the fermentation liquor, the molecular weight of one of the components was about 20.1 kDa. Fermentation liquor of B. laterosporus BL-21 could inhibit the growth of Alternaria solani and Bacillus cereus, so it was valuable for the research on its application potentiality in the field of biological control.