title: A simple and low-cost platform technology for producing pexiganan antimicrobial peptide in E. coli.
abstract: Antimicrobial peptides, as a new class of antibiotics, have generated tremendous interest as potential alternatives to classical antibiotics. However, the large-scale production of antimicrobial peptides remains a significant challenge. This paper reports a simple and low-cost chromatography-free platform technology for producing antimicrobial peptides in Escherichia coli (E. coli). A fusion protein comprising a variant of the helical biosurfactant protein DAMP4 and the known antimicrobial peptide pexiganan is designed by joining the two polypeptides, at the DNA level, via an acid-sensitive cleavage site. The resulting DAMP4(var)-pexiganan fusion protein expresses at high level and solubility in recombinant E. coli, and a simple heat-purification method was applied to disrupt cells and deliver high-purity DAMP4(var)-pexiganan protein. Simple acid cleavage successfully separated the DAMP4 variant protein and the antimicrobial peptide. Antimicrobial activity tests confirmed that the bio-produced antimicrobial peptide has the same antimicrobial activity as the equivalent product made by conventional chemical peptide synthesis. This simple and low-cost platform technology can be easily adapted to produce other valuable peptide products, and opens a new manufacturing approach for producing antimicrobial peptides at large scale using the tools and approaches of biochemical engineering.
authors.affiliation: The University of Queensland, Australian Institute for Bioengineering and Nanotechnology, St. Lucia, QLD 4072, Australia.
authors.forename: Chun-Xia
authors.initials: CX
authors.lastname: Zhao
authors.forename: Mirjana Dimitrijev
authors.initials: MD
authors.lastname: Dwyer
authors.forename: Alice Lei
authors.initials: AL
authors.lastname: Yu
authors.forename: Yang
authors.initials: Y
authors.lastname: Wu
authors.forename: Sheng
authors.initials: S
authors.lastname: Fang
authors.forename: Anton P J
authors.initials: AP
authors.lastname: Middelberg
mesh_terms: D000595:Amino Acid Sequence; D000890:Anti-Infective Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011232:Chemical Precipitation; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D007218:Industrial Microbiology / Q000191:economics / Q000379:methods; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D011993:Recombinant Fusion Proteins; C093828:pexiganan
keywords: antimicrobial peptide; chromatography-free; expression; precipitation; purification; recombinant