侧孢短芽孢杆菌BL-21抗菌蛋白的稳定性分析和分离纯化-中国农学通报.PDFVIP

中国农学通报 2017,33(15):43-48 Chinese Agricultural Science Bulletin 侧孢短芽孢杆菌BL-21抗菌蛋白的稳定性分析和分离纯化 张 淼，周立梅，徐德阳，杜春梅 （黑龙江大学生命科学学院，黑龙江省高校微生物重点实验室，哈尔滨150080） 摘 要：为明确侧孢短芽孢杆菌BL-21(Brevibacillus laterosp orus)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 kDa 。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。 关键词：侧孢短芽孢杆菌；抗菌物质；分离纯化 中图分类号：Q939.92 文献标志码：A 论文编号：cas StabilityAnalysis,IsolationandPurificationofAntimicrobialProteinFromBrevibacilluslaterosporus BL-21 ZhangMiao,ZhouLimei,XuDeyang,DuChunmei (KeyLaboratoryofMicrobiologyofCollegesofHeilongjiangProvince,CollegeofLifeSciences, HeilongjiangUniversity,Harbin150080) Abstract: Toclarifythetypeofantimicrobialsubstanceproducedby Brevibacillus laterosporus BL-21,the stabilityofthefermentationliquorwasanalyzed,andtheactiveingredientswereisolatedandpurifiedbasedon this.Thethermostability,pHstabilityandproteasestabilityofantimicrobialmetabolitesfrom B laterosporus BL-21 weredetectedbycylinderplatemethod.Theantimicrobialsubstanceswereisolatedandpurifiedby ammonium sulfate precipitation, CM-Sepharose Fast Flow ion exchange chromatography, gel filtration chromatography and SDS. The results showed that the antimicrobial substance produced by B laterosporusBL-21wasstablebelow50℃,itsantifungalactivitywasthebestatpH7-8,anditwassensitiveto protease.Severalantimicrobialcomponentswereisolatedfromthefermentationliquor,themolecularweightof oneofthecomponentswas