摘要
抗菌肽是目前最有希望的抗生素替代品,但是使用重组技术生产抗菌肽的策略大多步骤烦琐且价格昂贵,不利于抗菌肽的规模化生产。Oxysterlin1是一种新型的天蚕素抗菌肽,主要对革兰氏阴性菌有抗菌活性,具有较低的细胞毒性。文中利用一种简单经济的方法在大肠杆菌中实现Oxysterlin 1的表达和纯化。将Oxysterlin 1基因克隆到含有弹性蛋白样多肽Elastin-like polypeptide(ELP)和蛋白质内含肽(Intein)的载体中,构建重组表达质粒pET-ELP-I-Oxysterlin1。重组蛋白在大肠杆菌中主要以可溶性形式表达,进而通过简单的盐析和pH改变便可对目标小肽进行纯化。最终得到的Oxysterlin 1的产量约为1.2mg/L,抑菌试验显示出预期活性,为抗菌肽的规模化生产及深入研究其抑菌机理奠定基础。
Antimicrobial peptides are the most promising alternatives to antibiotics.However,the strategy of producing antimicrobial peptides by recombinant technology is complicated and expensive,which is not conducive to the large-scale production.Oxysterlin 1 is a novel type of cecropin antimicrobial peptide mainly targeting on Gram-negative bacteria and is of low cytotoxicity.In this study,a simple and cost-effective method was developed to produce Oxysterlin 1 in Escherichia coli.The Oxysterlin 1 gene was cloned into a plasmid containing elastin-like polypeptide(ELP)and protein splicing elements(intein)to construct the recombinant expression plasmid(pET-ELP-I-Oxysterlin 1).The recombinant protein was mainly expressed in soluble form in E.coli,and then the target peptide can be purified with a simple salting out method followed by pH changing.The final yield of Oxysterlin 1 was about 1.2 mg/L,and the subsequent antimicrobial experiment showed the expected antimicrobial activity.This study holds promise for large-scale production of antimicrobial peptides and the in-depth study of its antimicrobial mechanism.
作者
郭丽
刘化鑫
林瑛
Li Guo;Huaxin Liu;Ying Lin(College of Chemistry,Chemical Engineering and Biotechnology,Donghua University,Shanghai 201620,China)
出处
《生物工程学报》
CAS
CSCD
北大核心
2021年第8期2915-2923,共9页
Chinese Journal of Biotechnology
基金
上海市自然科学基金(No.19ZR1471000)
国家自然科学基金(No.31470836)资助。
