6×His-tag在抗菌肽表达纯化中的应用及其对抗菌肽活性的影响

Application of 6×His-tag in the Expression of Antibacterial Peptide and its Effect on the Antibacterial Activity

为方便抗菌肽的体外表达和纯化,以蝎子防御素为对象,在其编码基因末端加入6×His-tag,在酵母表达系统中表达,获得有活性的防御素Sd和融合蛋白Sd-His。进一步纯化、体外抑菌活性及热酸稳定性等试验发现,6×His-tag不但没有降低防御素的抑菌活性,反而有稍微的增强趋势。结果表明,6×His-tag可用于抗菌肽的融合表达且不影响其活性,为抗菌肽的表达纯化提供了一个更简便有效的方法。

For the convenience of expression and purification of antimicrobial peptides in vitro, the scorpion defensin gene（sd） and the gene fused with 6 × His-tag （sd-His） were reconstructed and expressed in P. pastoris. The two peptides were purified and then the antibacterial activity, thermal and acid stability were detected in Vitro. Surprisingly,the antibacterial activity of Sd-His had not been reduced but slightly enhanced. The results indicate that 6 × His-tag can be used for the fusion expression of antimicrobial peptide without affecting its activity. This study provides a simple and efficient method for the expression and purification of antimicrobial peptide.