基于短链抗菌肽Cn-AMP1的拼接肽合成及抗菌活性研究

Study on synthesis and antibacterial activity of splicing peptides based on short chain antibacterial peptide Cn-AMP1

目的以短链抗菌肽Cn-AMP1为基础拼接得到的抗菌肽C2和C4的抗菌活性和抗菌机制研究。方法应用生物信息学方法分析多肽的理化性质参数;使用圆二色光谱法测定其二级结构;使用CCK-8法测定其对BEAS-2B细胞毒性;通过抑菌实验和细菌细胞外膜实验,研究Cn-AMP1、C2、C4肽靶向细菌细胞膜的作用机制。结果圆二色光谱测定结果表明,在模拟膜环境中设计合成的C2和C4为α-螺旋结构;抗菌肽C2和C4相较于母肽Cn-AMP1具有良好的抗菌活性以及较低的细胞毒性;此外抗菌肽C2和C4与母肽Cn-AMP1相比都具有个更强的破坏细胞膜能力。结论C2和C4肽具有比Cn-AMP1肽更好的抗菌活性。

Objective To study the antibacterial activity and corresponding mechanism of antibacterial peptide C2 and C4 which synthesized based on the short sequence antibacterial peptide Cn-AMP1.Methods Bioinformatics analysis were applied to analyze the physicochemical properties and parameters of peptides,the secondary structure was determined by circular dichroism spectroscopy.The toxicity to BEAS-2B cells was determined by CCK-8 method,the mechanism of Cn-AMP1,C2,C4 peptides targeting bacterial cell membrane was studied through antimicrobial activity and bacterial outer membrane experiments.Results The results of circular dichroism spectrum indicate that the designed and synthesized C2 and C4 wereα-helix structure in the simulated membrane environment.The C2 and C4 peptides had stronger antimicrobial activity and lower cytotoxicity than the parent peptide.In addition,C2 and C4 peptides had good membrane breaking effects compared to the parent peptide Cn-AMP1.Conclusions C2 and C4 peptides have better antibacterial activity than Cn-AMP1 peptide.