Development of Elastin-Like Recombinamer Films with Antimicrobial Activity

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• 作者：Andr茅 da Costa ; Raul Machado ; Artur Ribeiro ; Tony Collins ; Viruthachalam Thiagarajan ; Maria Teresa Neves-Petersen ; Jos茅 Carlos Rodr铆guez-Cabello ; Andreia C. Gomes ; Margarida Casal
• 刊名：Biomacromolecules
• 出版年：2015
• 出版时间：February 9, 2015
• 年：2015
• 卷：16
• 期：2
• 页码：625-635
• 全文大小：597K
• ISSN：1526-4602

文摘

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.