title: Development of elastin-like recombinamer films with antimicrobial activity.
abstract: In the present work we explored the ABP-CM4 peptide properties from Bombyx mori for the creation of biopolymers with broad antimicrobial activity. An antimicrobial recombinant protein-based polymer (rPBP) was designed by cloning the DNA sequence coding for ABP-CM4 in frame with the N-terminus of the elastin-like recombinamer consisting of 200 repetitions of the pentamer VPAVG, here named A200. The new rPBP, named CM4-A200, was purified via a simplified nonchromatographic method, making use of the thermoresponsive behavior of the A200 polymer. ABP-CM4 peptide was also purified through the incorporation of a formic acid cleavage site between the peptide and the A200 sequence. In soluble state the antimicrobial activity of both CM4-A200 polymer and ABP-CM4 peptide was poorly effective. However, when the CM4-A200 polymer was processed into free-standing films high antimicrobial activity against Gram-positive and Gram-negative bacteria, yeasts and filamentous fungi was observed. The antimicrobial activity of CM4-A200 was dependent on the physical contact of cells with the film surface. Furthermore, CM4-A200 films did not reveal a cytotoxic effect against both normal human skin fibroblasts and human keratinocytes. Finally, we have developed an optimized ex vivo assay with pig skin demonstrating the antimicrobial properties of the CM4-A200 cast films for skin applications.
authors.affiliation: CBMA (Centre of Molecular and Environmental Biology), Department of Biology, University of Minho , Campus de Gualtar, 4710-057 Braga, Portugal.
authors.forename: André
authors.initials: A
authors.lastname: da Costa
authors.forename: Raul
authors.initials: R
authors.lastname: Machado
authors.forename: Artur
authors.initials: A
authors.lastname: Ribeiro
authors.forename: Tony
authors.initials: T
authors.lastname: Collins
authors.forename: Viruthachalam
authors.initials: V
authors.lastname: Thiagarajan
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Neves-Petersen
authors.forename: José Carlos
authors.initials: JC
authors.lastname: Rodríguez-Cabello
authors.forename: Andreia C
authors.initials: AC
authors.lastname: Gomes
authors.forename: Margarida
authors.initials: M
authors.lastname: Casal
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000737:chemistry* / Q000494:pharmacology*; D012831:Bombyx; D002460:Cell Line; D004337:Drug Carriers / Q000737:chemistry / Q000494:pharmacology; D004549:Elastin / Q000737:chemistry* / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects / Q000502:physiology; D006801:Humans; D012869:Skin Absorption / Q000187:drug effects / Q000502:physiology; D013552:Swine
chemical_list: D000890:Anti-Infective Agents; D004337:Drug Carriers; D004549:Elastin