人类抗菌肽hepcidin在毕赤酵母中的分泌表达和纯化  

Secretory Expression and Purification of a Human Hepcidin in P. pastoris

作　　者：Farzana Rashid Ijaz Ali 袁其朋[1] 孙新晓[1] 李文进[1] 李飞[1] 

机构地区：[1]北京化工大学生命科学与技术学院,北京100029

出　　处：《生物技术通报》2009年第9期121-124,共4页Biotechnology Bulletin

基　　金：国家自然科学基金(20576010)

摘　　要：报道了一种低分子量的人类抗菌肽hepcidin基因的克隆、表达以及纯化。试验证明在毕赤酵母中能成功分泌有活性的hepcidin,hepcidin在重组菌株中的表达量约为3 mg/L,Western blot显示2.2 kD的重组hepcidin条带,重组hepcidin通过凝胶过滤及反向高效液相色谱纯化,质谱验证,重组蛋白对金黄色葡萄球菌和枯草芽孢杆菌表现出抗菌活性。Expression and purification of small peptides have always been problematic due to enzymatic degradation and many other technical problems. We reported cloning and expression of a low molecular weight human antimicrobial peptide hepcidin（ Hepc, 20 amino acids） into pPIC9K transformed into P. pastoris GSllS. The study revealed that active hepcidin peptide can be successfully expressed in this methylotrophic yeast. Hepcidin protein expressed in recombinant strains was about 3 mg/L. Peptide expression was detected by immuno blotting. Recombinant hepc 20 was purified through reverse - phase HPLC column and characterized by mass spectrometry and amino acid sequencing. It also exhibited antibacterial activity against S. aureus and Bacillus subtilis. Such a research work is being presented for the first time and it is hoped that this will provide a basis for the mass production of Hepc 20.

关 键 词：抗菌肽 HEPCIDIN 20 铁 毕赤酵母 分泌表达 

分 类 号：Q78[生物学—分子生物学]