Yi Lanhua, Dang Ying, Wu Jingli, Zhang Lihui, Liu Xiaojiao, Liu Bianfang, Zhou Yuan, Lu Xin
College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi Province 712100, China.
College of Food Science and Engineering, Northwest A&F University, Yangling, Shaanxi Province 712100, China.
J Dairy Sci. 2016 Sep;99(9):7002-7015. doi: 10.3168/jds.2016-11166. Epub 2016 Aug 8.
The growing emergence of antibiotic-resistant bacteria in the food industry needs to be controlled with effective antimicrobials. In this study, bacteriocin MN047 A (BMA) was found to have antibacterial activity against multidrug-resistant bacteria. It was produced by Lactobacillus crustorum MN047, which was first isolated from koumiss, a traditional fermented dairy product from Xinjiang Autonomous Region, China. It was purified by ammonium sulfate precipitation, ion-exchange chromatography, and reversed-phase chromatography. It had a low molecular mass of 1,770.89 Da according to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and the sequence was identified as QLPWQILGIVAGMFQA by liquid chromatography-tandem mass spectrometry analysis and MASCOT searching. It was proteinaceous in nature: the bacteriocin was digested by protease but not by α-amylase or lipase. It showed broad pH toleration (pH 2-11), good thermostability, and good storage stability. It had a broad inhibitory spectrum, including both gram-positive and gram-negative bacteria. Growth curve and time-kill kinetics indicated that it was bactericidal to the indicator strains, and this finding was verified by scanning electron microscope and transmission electron microscope after treatment with BMA. As well, BMA halted the growth of Staphylococcus aureus and Escherichia coli in the G1 and G2/M phases according to cell-cycle analysis by flow cytometry, indicating that BMA had comprehensive inhibitory effects against foodborne pathogens.
食品工业中抗生素耐药细菌的不断出现需要用有效的抗菌剂加以控制。在本研究中,发现细菌素MN047 A(BMA)对多重耐药细菌具有抗菌活性。它由酪样乳杆菌MN047产生,该菌株最初从中国新疆自治区的传统发酵乳制品马奶酒中分离得到。通过硫酸铵沉淀、离子交换色谱和反相色谱对其进行纯化。根据基质辅助激光解吸/电离飞行时间质谱分析,其分子量较低,为1770.89 Da,通过液相色谱-串联质谱分析和MASCOT搜索,其序列被鉴定为QLPWQILGIVAGMFQA。它本质上是蛋白质类:该细菌素可被蛋白酶消化,但不能被α-淀粉酶或脂肪酶消化。它表现出广泛的pH耐受性(pH 2 - 11)、良好的热稳定性和良好的储存稳定性。它具有广泛的抑制谱,包括革兰氏阳性菌和革兰氏阴性菌。生长曲线和时间杀灭动力学表明它对指示菌株具有杀菌作用,在用BMA处理后通过扫描电子显微镜和透射电子显微镜验证了这一发现。同样,根据流式细胞术的细胞周期分析,BMA使金黄色葡萄球菌和大肠杆菌的生长在G1期和G2/M期停滞,表明BMA对食源性病原体具有全面的抑制作用。