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DEFB113为人β-防御素家族新成员,其生物学功能尚不清楚。为了探索一种可以较低成本和较高产量表达活性DEFB113多肽的方法,将编码DEFB113成熟肽的序列克隆到载体pTWIN1上,并转化至E.coliBL21(DE3)中进行表达。结果表明融合蛋白主要以可溶形式存在;重组载体中融合标签的几丁质结合域(CBD)和内含肽(intein)分别使亲和层析纯化和融合标签自剪切一步完成。最终得到DEFB113多肽的产率为6～7 mg/LLB。经活性鉴定,该重组DEFB113多肽对于S.aureus,E.coli K12D31和C.albicans均表现出较强的抑制作用;另外DEFB113无明显的溶血活性,是一种较理想的多肽抗生素。该研究为进一步研究DEFB113的生理功能以及其它人β-防御素结构与功能奠定了基础。
DEFB113 is a new member of the human β-defensin family and its biological function is unclear. To explore a method to express active DEFB113 polypeptide at lower cost and with higher yield, the sequence encoding the mature peptide of DEFB113 was cloned into vector pTWIN1 and transformed into E. coli BL21 (DE3) for expression. The results showed that the fusion protein mainly exists in soluble form. The fusion tagged chitin binding domain (CBD) and intein in the recombinant vector make the purification and fusion tag purification by affinity chromatography in one step, respectively. The final yield of DEFB113 polypeptide was 6-7 mg / LLB. The activity of the recombinant DEFB113 polypeptide showed a strong inhibitory effect on S.aureus, E. coli K12D31 and C.albicans. In addition, DEFB113 had no obvious hemolysis activity and was an ideal polypeptide antibiotic. This study lays the foundation for further study on the physiological function of DEFB113 and the structure and function of other human β-defensins.