Plant Biotechnology and Plant Breeding, Ferdowsi University of Mashhad, Iran.
Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran.
Microb Pathog. 2018 Sep;122:73-78. doi: 10.1016/j.micpath.2018.06.012. Epub 2018 Jun 8.
Over the last decades, poultry industry faced to the rapid emergence of multidrug-resistant bacteria as a global concern. Antimicrobial peptide (AMPs) known as potential antibiotic alternative and were considered as a new antimicrobial agent. Current methods of production and purification of AMPs have several limitations such as: costly, time-consuming and killing the producing host cells in recombinant form. In the present study, a chimeric peptide derived from camel lactoferrin was produced in Escherichia coli periplasmic space using a pET-based expression system and its antibacterial activity was determined on some avian pathogens in vitro. A carboxy-terminal polyhistidine tag was used for purification by Ni affinity chromatography with an average yield of 0.42 g/L. The His-tagged chimeric peptide showed different range of antimicrobial activity against clinically isolated avian pathogens with low chicken blood hemolysis activity and high serum stability. Overall, the results of this investigation showed the recombinant chimeric peptide was successfully expressed in pET-based expression system and could be considered as a proper alternative for some currently used antibiotics in poultry industry and drugs veterinary medicine.
在过去几十年中，家禽业面临着多药耐药细菌的迅速出现，这是一个全球性的关注问题。抗菌肽 (AMPs) 作为潜在的抗生素替代品而备受关注，并被认为是一种新型抗菌药物。目前 AMPs 的生产和纯化方法存在一些局限性，例如：成本高、耗时且会杀死重组形式的产生宿主细胞。在本研究中，使用基于 pET 的表达系统在大肠杆菌周质空间中生产了源自骆驼乳铁蛋白的嵌合肽，并在体外测定了其对一些禽源病原体的抗菌活性。羧基末端多组氨酸标签用于通过 Ni 亲和层析进行纯化，平均产率为 0.42 g/L。该 His 标记的嵌合肽对临床分离的禽源病原体表现出不同范围的抗菌活性，具有低鸡血液溶血活性和高血清稳定性。总体而言，这项研究的结果表明，重组嵌合肽已成功在基于 pET 的表达系统中表达，可作为家禽业和兽医药中一些目前使用的抗生素的替代品。
