Study on Refolding and Purification of Plectasin MP1106

doi:10.13560/j.cnki.biotech.bull.1985.2017-0287

Abstract

Abstract: This work aims to establish an efficient and simple Escherichia coli expression system for the antimicrobial peptide,plectasin MP1106. The recombinant MP1106 fusion protein was obtained by genetic fusion with the artificially designed bio-surfactant,DAMP4. Then,the recombinant DAMP4-MP1106 was expressed in E. coli and the protein was isolated,moreover the formation of the disulfide bonds within the protein was identified. The results showed that the recombinant DAMP4-MP1106 was over-expressed in inclusion body. The protein was obtained by the Ni2+-NTA chromatography under the denature condition. The total yield of the fusion protein after the first chromatography was 118 mg/L in flask and the purity was 94.7%. The refolding buffer condition was screened by 96-well plate and the soluble recombinant DAMP4-MP1106 was obtained by a refolding procedure. The soluble recombinant DAMP4-MP1106 was cleaved by TEV protease and further purified by another Ni2+-NTA column. The purity of final MP1106 was 99% and the recovery of the protein was 38.4%. The comparison of the behaviour of the plectasin derivative,MP1106 with the standard plectasin on electrophoresis indicated that the disulfide bonds in MP1106 were folded correctly. In conclusion,the established E. coli expression system for the plectasin derivative MP1106 was efficient and successful.

Key words: antimicrobial peptide, plectasin, protein fusion, refolding, protein purification

WANG Peng-ju,TAN Huan-bo,SU Wen-cheng,ZHANG Wen-yu,ZOU Pei-jian. Study on Refolding and Purification of Plectasin MP1106[J]. Biotechnology Bulletin, 2017, 33(9): 94-100.

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