摘要

通过生物信息学方法从18575个ORF中分离出潜在的细菌素基因。它被命名为pln1，并被克隆到pET32a中。然后，它在大肠杆菌BL21（DE3）中表达为硫氧还蛋白-Pln1融合蛋白。用Ni-NTA纯化融合蛋白，用肠激酶去除硫氧还蛋白。最后，使用阳离子亲和柱纯化Pln1。融合和切割的Pln1肽的产量分别为100-110mg / l和9-11mg / l。 Pln1在酸性环境中和低于60℃的温度下稳定，但在碱性条件下和蛋白酶处理下易于降解。切割并纯化的Pln1对革兰氏阳性菌例如藤黄微球菌CMCC 63202，表皮葡萄球菌，乳酸乳球菌NZ3900，副干酪乳杆菌CICC 20241和无害李斯特菌CICC 10417显示出强的抗微生物活性。特别地，Pln1具有更好的抗甲氧西林 - 耐药表皮葡萄球菌（MRSE）比乳链菌肽，因此提供了一种有吸引力的方法来抵抗细菌抗生素耐药性。

A potential bacteriocin gene was isolated from 18575 ORFs by bioinformatics methods. It was named pln1, and cloned into pET32a. Then, it was expressed as a thioredoxin-Pln1fusion protein in Escherichia coli BL21 (DE3). The fusion protein was purified by Ni-NTA, and thioredoxin was removed by enterokinase. Finally, Pln1 was purified using a cation affinity column. The yields of fused and cleaved Pln1 peptides were 100–110 mg/l and 9–11 mg/l, respectively. Pln1 was stable in an acidic environment and at temperatures below 60 °C, but was easily degraded under alkaline conditions and by protease treatment. The cleaved and purified Pln1 showed strong antimicrobial activity against gram-positive bacteriasuch as Micrococcus luteus CMCC 63202, Staphylococcus epidermidis, Lactococcus lactisNZ3900, Lactobacillus paracasei CICC 20241, and Listeria innocua CICC 10417. In particular, Pln1 had a better activity against methicillin-resistant S. epidermidis (MRSE) thannisin, thereby offering an attractive approach to counter bacterial antibiotic resistance.

https://www.sciencedirect.com/science/article/pii/S1046592815301005