The safe enterocin DD14 is a leaderless two-peptide bacteriocin with anti-Clostridium perfringens activity
The safe enterocin DD14 is a leaderless two-peptide bacteriocin with anti-Clostridium perfringens activity
复制标题
DOI：
10.1016/j.ijantimicag.2016.11.016
复制
发表时间：
2017-03-01
影响因子：
10.8
通讯作者：
Drider, Djamel
中科院分区：
文献类型：
--
作者：
Caly, Delphine L.;Chevalier, Mickael;Drider, Djamel
粪肠球菌 14 是一种先前从胎便中分离出来的菌株，当在琼脂平板上共培养时，显示出针对四种产气荚膜梭菌分离株的活性。抗梭菌活性归因于肠菌素 DD14 的产生，随后对其进行了纯化。确定肠菌素DD14对一种保藏菌株和一种临床产气荚膜梭菌的最低抑菌浓度(MIC)为50μg/mL。此外，使用肠上皮细胞系IPEC-1，显示粪肠球菌14在接触24小时后不具有细胞毒性，并且当IPEC-1细胞与纯肠菌素DD14孵育4小时时未观察到细胞毒性。使用质谱法对 Enterocin DD14 进行了表征，结果显示它由两种大小分别为 5200.74 Da 和 5206.41 Da 的小蛋白组成。两种肽（DD14A和DD14B）具有高度相似的氨基酸序列并且没有信号肽，这将肠菌素DD14归类为IIb类无前导序列的二肽细菌素。对编码 DD14A 和 DD14B 的基因进行了测序，结果显示与之前描述的其他肠菌素 MR10A 和 MR10B 100% 相同，而生产菌株则不同。因此，目前的体外研究支持该粪肠球菌 14 菌株和/或其纯化的肠菌素 DD14 作为推定的抗梭菌的潜力。鸡体内的产气荚膜梭菌化合物。 (C) 2017 年由 Elsevier B.V. 出版
Enterococcus faecalis 14, a strain previously isolated from meconium, displayed activity against four Clostridium perfringens isolates when co-cultured on agar plates. The anti-Clostridium activity was ascribed to the production of enterocin DD14, which was subsequently purified. The minimum inhibitory concentration (MIC) of enterocin DD14 against one collection strain and one clinical C. perfringens strain was determined at 50 mu g/mL. Furthermore, using the intestinal epithelial cell line IPEC-1, it was shown that E. faecalis 14 was not cytotoxic after 24 h of contact, and no cytotoxicity was observed when IPEC-1 cells were incubated with pure enterocin DD14 for 4 h. Enterocin DD14 was characterised using mass spectrometry and was shown to consist of two small proteins of 5200.74 Da and 5206.41 Da, respectively. The two peptides (DD14A and DD14B) have highly similar amino acid sequences and no signal peptide, which classifies enterocin DD14 as a class IIb leaderless two-peptide bacteriocin. The genes encoding DD14A and DD14B were sequenced and were shown to be 100% identical to other previously described enterocins MR10A and MR10B, in contrast to the producing strains, which are different. Consequently, the present in vitro study supports the potential of this E. faecalis 14 strain and/or its purified enterocin DD14 as putative anti-C. perfringens compounds in chickens. (C) 2017 Published by Elsevier B.V.