Abriouel Hikmate, Valdivia Eva, Martínez-Bueno Manuel, Maqueda Mercedes, Gálvez Antonio
Departamento de Microbiología, Facultad de Ciencias, Universidad de Granada, Granada, Spain.
J Microbiol Methods. 2003 Dec;55(3):599-605. doi: 10.1016/s0167-7012(03)00202-1.
Production of enterocin AS-48 by Enterococcus faecalis A-48-32 was compared between standard and high-cell density batch fermentations. In high-cell density cultures, bacteriocin production was 2.47-fold higher, provided that the pH was controlled during the fermentation. A two-step procedure for recovery of milligram quantities of purified bacteriocin was developed, based on adsorption of the bacteriocin on Carboxymethyl Sephadex CM-25 followed by reversed-phase chromatography on a semi-preparative column. The purified bacteriocin was active on all the Gram-positive bacteria tested (for example, species of Bacillus, Paenibacillus, Staphylococcus, and Listeria). Strains E. coli U-9, E. coli CECT 102, E. coli CECT 104, E. coli CECT 432, E. coli CECT 543, E. coli CECT 877 and Shigella sonnei CECT 542 were sensitive, while seven other E. coli strains as well as Salmonella choleraesuis CECT 722, S. choleraesuis CECT 916, Enterobacter cloacae CECT 194 and Aeromonas hydrophila CECT 398 were resistant.
比较了粪肠球菌A-48-32在标准分批发酵和高细胞密度分批发酵中肠球菌素AS-48的产生情况。在高细胞密度培养中,只要在发酵过程中控制pH值,细菌素的产量就会高出2.47倍。基于细菌素在羧甲基葡聚糖凝胶CM-25上的吸附以及随后在半制备柱上的反相色谱法,开发了一种两步法来回收毫克量的纯化细菌素。纯化的细菌素对所有测试的革兰氏阳性菌(例如芽孢杆菌属、类芽孢杆菌属、葡萄球菌属和李斯特菌属的菌种)都有活性。大肠杆菌U-9、大肠杆菌CECT 102、大肠杆菌CECT 104、大肠杆菌CECT 432、大肠杆菌CECT 543、大肠杆菌CECT 877和宋内志贺氏菌CECT 542菌株敏感,而其他七种大肠杆菌菌株以及猪霍乱沙门氏菌CECT 722、猪霍乱沙门氏菌CECT 916、阴沟肠杆菌CECT 194和嗜水气单胞菌CECT 398具有抗性。