摘要
试验旨在探究地衣芽孢杆菌38(Bacillus licheniformis 38,B38)对产气荚膜梭菌(Clostridium perfringens,CP)的抑菌性能,以期为新型替抗产品的开发提供基础。采用抑菌圈法对48株芽孢杆菌经过两次筛选,获得一株对CP抑菌效果最好的芽孢杆菌,测定其最小抑菌效价、抑菌产物生成曲线、抑菌谱和代谢产物的稳定性,并通过硫酸铵沉淀法对抗菌物质进行粗提取。结果表明:B38对CP的抑菌圈直径最大可达31.14 mm,其发酵上清液的最小抑菌效价为64 AU/mL,主要对革兰氏阳性菌发挥抑菌作用;B38代谢产物的抑菌活性在6~<12 h呈指数增长,在12~<24 h保持稳定;B38代谢产物经胃蛋白酶、胰蛋白酶、蛋白酶K和过氧化氢酶处理后,抑菌活性均大于87.78%,对温度和pH具有良好的耐受性;B38代谢产物中的抑菌物质可以被80%硫酸铵完全沉淀,通过该方法获得的抗菌粗品的MIC为6.25 mg/mL。试验通过抑菌圈法筛选得到了一株对CP具有优良抑菌性能的菌株B38,其发酵产物对蛋白酶、温度和pH具有优良的耐受能力。
The experiment aims to explore the bacteriostatic properties of Bacillus licheniformis B38(B38)against Clostridium perfringens(CP),so as to provide a basis for the development of new alternative antibiotic products.A total of 48 strains of Bacillus were screened twice with the method of bacteriostasis circle,and one strain of Bacillus with the best bacteriostasis effect against CP was obtained.Its minimum bacteriostasis potency,bacteriostasis product generation curve,bacteriostasis spectrum and stability of metabolites were determined,and the antibacterial substances were roughly extracted by ammonium sulfate precipitation method.The results showed that the maximum diameter of the bacteriostatic zone of B38 against CP was 31.14 mm,and the minimum bacteriostatic potency of its fermentation supernatant was 64 AU/mL,mainly playing a bacteriostatic role against gram-positive bacteria;The antibacterial activity of B38 metabolites increased exponentially from 6 to<12 hours and remained stable from 12 to<24 hours;after being treated with pepsin,trypsin,proteinase K and catalase,B38 metabolites have antibacterial activities greater than 87.78%,and have good tolerance to temperature and pH;the antibacterial substances in B38 metabolites can be completely precipitated by 80%ammonium sulfate,and the MIC of the crude antibacterial product obtained by this method is 6.25 mg/mL.In this paper,a strain B38 with excellent bacteriostasis against CP was screened by the method of bacteriostasis ring.Its fermentation product has excellent tolerance to protease,temperature and pH.
作者
杨梦瀚
苗华彪
宋婧楠
赵香帅
陈紫娴
吴倩
黄遵锡
YANG Menghan;MIAO Huabiao;SONG Jingnan;ZHAO Xiangshuai;CHEN Zixian;WU Qian;HUANG Zunxi(College of Life Sciences,Yunnan Normal University,Yunnan Kunming 650500,China;Engineering Research Center for Sustainable Development and Utilization of Bioenergy,Ministry of Education,Yunnan Kunming 650500,China)
出处
《饲料工业》
北大核心
2023年第15期92-99,共8页
Feed Industry
基金
国家自然科学基金项目[31960131]
中央引导地方科技发展资金项目[202107AC110002]
大学生创新创业训练计划项目[202110681012、S202110681075]。
关键词
芽孢杆菌
地衣芽孢杆菌
产气荚膜梭菌
耐受
抑菌性能
Bacillus
Bacillus licheniformis
Clostridium perfringens
tolerance
antibacterial activity
