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前期研究表明枯草芽孢杆菌HAINUP40能够有效拮抗罗非鱼无乳链球菌,是潜在的益生菌。为了进一步优化其培养条件,并探索其其抑菌机理,本研究通过普通平板扩散法和单因素变量控制法,探究了培养条件的改变对菌株HAINUP40发酵的影响,同时确定发酵上清液抑菌活性成分及其稳定性。结果表明菌株HAINUP40的最宜培养条件为:普通淡水培养基,pH值为7.0～7.5,温度34℃,培养时长24 h,转速为50～150 r/min。HAINUP40发酵上清液中抑菌活性物质可以被80%(NH4)_2SO4溶液析出,对蛋白酶K敏感,不耐高温,pH<4或>8时失去抑菌活性,可以耐受短时间(<180 s)紫外线照射。综上证明枯草芽孢杆菌HAINUP40所分泌的抑菌活性物质为大分子蛋白质,且非小分子抗菌肽。本研究为枯草芽孢杆菌HAINUP40在生物防治方面的大规模应用提供了重要的参考依据。
Abstract：Prior studies revealed that Bacillus subtilis HAINUP40 could effectively inhibit Streptococcus agalactiae pathogen in tilapia and was considered as a promising probiotic candidate. To optimize the culture conditions of HAINUP40, as well as illustrating its antibacterial mechanism, we studied the effects of different culture conditions on the fermentation of HAINUP40 and the antibacterial substances in the fermentation supernatant of HAINUP40 using agarose plate diffusion experiment and the single factor experiment. Results showed that the optimal conditions for HAINUP40 fermentation were: Freshwater medium, pH at 7.0～7.5, temperature at 34℃, culture time for24 h, rotation speed at 50～150 r/min. The antibacterial substances could be precipitated with 80%(NH4)_2SO4, and they were sensitive to proteinase K and high temperature. They lost their antibacterial activities when pH<4 or pH>8.They were tolerant to short-term(<180 s) ultraviolet irradiation. Based on above results, we inferred the active antibacterial substances of Bacillus subtilis HAINUP40 were high molecular weight proteins and not lipopeptide.This study will provide an important reference for the large-scale application of Bacillus subtilis HAINUP40 in the biological control of bacterial pathogens.
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基本信息:
DOI：10.13417/j.gab.039.002638
中图分类号:S476.1
引用信息:
[1]吴越,王世锋,陈鑫,等.枯草芽孢杆菌HAINUP40的培养条件及其抑菌物质分析[J].基因组学与应用生物学,2020,39(06):2638-2645.DOI:10.13417/j.gab.039.002638.
基金信息:
国家自然科学基金(31860739; 31560725)资助
2019-03-16
2019-03-16
2019-03-16