北京化工大学学报（自然科学版）
北京化工大學學報（自然科學版）
북경화공대학학보（자연과학판）
JOURNAL OF BEIJING UNIVERSITY OF CHEMICAL TECHNOLOGY(NATURAL SCIENCE EDITION)
2010年
2期
104-108
,共5页
Farzana RASHID%袁其朋%孙新晓%李飞
Farzana RASHID%袁其朋%孫新曉%李飛
Farzana RASHID%원기붕%손신효%리비
抗菌肽%Hepc 20%铁%毕赤酵母
抗菌肽%Hepc 20%鐵%畢赤酵母
항균태%Hepc 20%철%필적효모
antimicrobial peptide%hepcidin 20%iron%Pichia pastoris
构建了Hepc 20的毕赤酵母表达载体,在毕赤酵母中能成功表达有活性的Hepc 20.优化了菌株的培养诱导条件,结果表明BMMY培养基是Hepc 20表达和重组菌株生长的最佳培养基,甲醇诱导终体积分数0.5%,在此条件下Hepc 20在重组菌株中的表达量约为3.6mr/L.Western blot检测显示的22000处的条带为重组Hepc-20条带;ELISA验证表明重组Hepc 20可以跟抗体特异结合,琼脂扩散法抗菌实验表明重组Hepc 20对金黄色葡萄球菌和枯草芽孢杆菌表现出抗菌活性.
構建瞭Hepc 20的畢赤酵母錶達載體,在畢赤酵母中能成功錶達有活性的Hepc 20.優化瞭菌株的培養誘導條件,結果錶明BMMY培養基是Hepc 20錶達和重組菌株生長的最佳培養基,甲醇誘導終體積分數0.5%,在此條件下Hepc 20在重組菌株中的錶達量約為3.6mr/L.Western blot檢測顯示的22000處的條帶為重組Hepc-20條帶;ELISA驗證錶明重組Hepc 20可以跟抗體特異結閤,瓊脂擴散法抗菌實驗錶明重組Hepc 20對金黃色葡萄毬菌和枯草芽孢桿菌錶現齣抗菌活性.
구건료Hepc 20적필적효모표체재체,재필적효모중능성공표체유활성적Hepc 20.우화료균주적배양유도조건,결과표명BMMY배양기시Hepc 20표체화중조균주생장적최가배양기,갑순유도종체적분수0.5%,재차조건하Hepc 20재중조균주중적표체량약위3.6mr/L.Western blot검측현시적22000처적조대위중조Hepc-20조대;ELISA험증표명중조Hepc 20가이근항체특이결합,경지확산법항균실험표명중조Hepc 20대금황색포도구균화고초아포간균표현출항균활성.
A recombinant Pichia pastoris strain for Hepc 20 production has been constructed and an active hepcidin peptide was successfully expressed in methylotrophic yeast.A buffered methanol complex(BMMY)medium was found to be optimal for the hepcidin protein expression and growth of the recombinant strains,and when the final volume fraction of methanol was 0.5%,the expression level of hepcidin protein reached the maximum value of 3.6 mg/L.Western blotting showed the 22000 band of the recombinant hepcidin protein whilst enzyme-linked immu-nosorbent(ELISA)assay showed that the recombinant Hepc 20 peptide can combine with specific antibodies,and the agar diffusion technique showed that Hepc 20 exhibited antibacterial activity against S.aureus and B.subtilis.
