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High‐Level Expression of a Soluble Functional Antimicrobial Peptide, Human β‐Defensin 2, in Escherichia coli
Author(s) -
Xu Zhinan,
Peng Li,
Zhong Zhixia,
Fang Xiangming,
Cen Peilin
Publication year - 2006
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp0502680
Subject(s) - escherichia coli , defensin , chemistry , affinity chromatography , fusion protein , recombinant dna , beta defensin , antimicrobial , ultrafiltration (renal) , solubility , chromatography , antimicrobial peptides , peptide , biochemistry , enzyme , organic chemistry , gene
In this work, taking human β‐defensin‐2 (HBD2) as a demonstrative molecule, the strategies for high efficient production of functional human β‐defensins in E. coli were studied. Fusion mature HBD2 (TrxA‐mHBD2) showed high solubility and productivity without the need for lowering the cultivation temperature. The solubility of target fusion protein could attain 81.3% even at 37 °C with a volumetric productivity as high as 235 mg/L in a rich medium MBL at the same temperature and reached 346 mg/L at 28 °C. The His‐Tag in the fusion protein enabled the application of affinity chromatography separation to obtain high purity of the overexpressed recombinant fusion protein. After digestion by enterokinase, purification via cationic exchange chromatography, and desalting by ultrafiltration, mature HBD2 product was obtained with a purity of 95% in an overall recovery of 29.2%. The antimicrobial activity of the recombinant mature HBD2 and the influence factors were tested using E. coli K12D31 as a sensitive strain.