摘要
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 recently identified human β-defensin,while its biological functions remain unclear.With the purpose of providing a low-cost and high-yielding approach to express DEFB113 for the determination of its bioactivities,the sequence encoding the mature peptide of DEFB113 was inserted into pTWIN1 and the recombinant transformed vector into E.coli BL21（DE3） for the recombinant expression.Most of the fusion proteins were expressed in a soluble form.Moreover,the chitin-binding domain（CBD） and the intein in the fusion tag facilitated the affinity chromatography and the auto-cleavage of the fusion tag,respectively.The resulting DEFB113 peptide was produced with a yield of 6.4 mg/L LB.The purified DEFB113 peptide exhibited potent antimicrobial activities against E.coli,S.aureus and C.albicans.In addition,the recombinant DEFB113 peptide caused little if any hemolysis towards human erythrocytes.In this respect,DEFB113 might exist as a promising peptide antibiotic.These attempts have laid the basis for further study on the physiological roles of DEFB113 as well as on the structure-function relationships of other DEFBs.
出处
《药物生物技术》
CAS
CSCD
2012年第1期1-5,共5页
Pharmaceutical Biotechnology
基金
上海市优秀学科带头人计划(A类)(No.09XD1405100)
