侧孢短芽孢杆菌BL-21抗菌蛋白的稳定性分析和分离纯化

doi:10.11924/j.issn.1000-6850.casb16110071

中国农学通报 ›› 2017, Vol. 33 ›› Issue (15): 43-48.doi: 10.11924/j.issn.1000-6850.casb16110071

所属专题：生物技术

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）

收稿日期:2016-11-15 修回日期:2017-05-05 接受日期:2017-01-16 出版日期:2017-05-26 发布日期:2017-05-26
通讯作者: 杜春梅
基金资助:
国家自然科学基金“侧孢短芽孢杆菌与枯草芽孢杆菌共培养抗菌活性增强的生态学机理”(31370511)；黑龙江省自然科学基金“侧孢芽孢杆菌与枯草芽孢杆菌共培养代谢产物的研究”(C201203)；黑龙江大学微生物学重点实验室开放项目“侧孢短芽孢杆菌与枯草芽孢杆菌共培养的生态学关系”(2012MOI-1)。

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

Zhang Miao, Zhou Limei, Xu Deyang, Du Chunmei

(Key Laboratory of Microbiology of Colleges of Heilongjiang Province, College of Life Sciences, Heilongjiang University, Harbin 150080)

Received:2016-11-15 Revised:2017-05-05 Accepted:2017-01-16 Online:2017-05-26 Published:2017-05-26

摘要/Abstract

摘要： 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 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: 枣树, 枣树, 枣股, 枣吊, 生长发育规律, 综合指数

Abstract: To clarify the type of antimicrobial substance produced by Brevibacillus laterosporus BL-21, the stability of the fermentation liquor was analyzed, and the active ingredients were isolated and purified based on this. The thermostability, pH stability and protease stability of 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℃, its antifungal activity was the best at pH 7-8, and it was sensitive to protease. Several antimicrobial components were 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.

中图分类号:

Q939.92

张淼,周立梅,徐德阳,杜春梅. 侧孢短芽孢杆菌BL-21抗菌蛋白的稳定性分析和分离纯化[J]. 中国农学通报, 2017, 33(15): 43-48.

Zhang Miao,Zhou Limei,Xu Deyang and Du Chunmei. Stability Analysis, Isolation and Purification of Antimicrobial Protein From Brevibacillus laterosporus BL-21[J]. Chinese Agricultural Science Bulletin, 2017, 33(15): 43-48.

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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

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