Creation of antimicrobial biopolymers by the use of recombinant DNA technology

[Excerpt] The spread of antimicrobials resistant microorganisms has triggered the search for new ways to treat infections. One of these ways is the creation of antimicrobial devices and surfaces that kill or prevent the spread of microorganisms. In the present work we explored the properties of different antimicrobial peptides (AMPs) for the creation of biopolymers with broad antimicrobial activity. Antimicrobial recombinant protein-based polymers (rPBPs) were designed by cloning the DNA sequence coding for the different AMPs in frame with the N-terminus of the elastin-like recombinamer consisting of 200 repetitions of the pentamer VPAVG, here named A200. [...]This work was supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through the COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI). By the Spanish Minister of Economy and Competitiveness (MAT2012-38043-C02-01) and Junta de Castilla y León-JCyL (VA152A12-2 and VA155A12-2), Spain. AC and RM, acknowledge FCT for SFRH/BD/75882/2011 and SFRH/BPD/86470/2012 grants, respectively

Antibacterial protein-based fibres: combining recombinant DNA technology with electrospinning

[Excerpt] With the increasing healthcare-associated infections and antibiotic-resistant microorganisms there is a demand not only for new antimicrobial compounds but also for antimicrobial materials. Genetically engineered protein polymers functionalized with bioactive domains offer potential as multifunctional versatile materials for biomedical use. The present work describes the fabrication and characterization of antimicrobial fibre mats comprising the antimicrobial elastin-like recombinamer CM4-A200 [1]. [...]This work was supported by the strategic programme UID/BIA/04050/2013 (POCI-01- 0145-FEDER- 007569) through FCT I.P. and by ERDF through COMPETE2020 - POCI. The authors are grateful for funding from FCT (project “FunBioPlas” ERA-IB-2-6/0004/2014) and a fellowship to RM (SFRH-BPD/86470/2012). The authors also thank support from the COST Action MP1206

Electrospun fibres of an elastin-like polymer functionalized with an antimicrobial domain

This work describes the production and characterization of nanofibres of a functionalized elastin-like recombinamer (ELR). The polymer was functionalized with an antimicrobial peptide domain by means of recombinant DNA technology and processed by electrospinning. The electrospun fibres were characterized for their morphology, physical-chemical, antimicrobial and cytotoxicity properties. The electrospun membranes showed no cytotoxicity against skin-related cell lines, suggesting the potential applicability of these materials for skin tissue engineering.This work was supported by FEDER through POFC – COMPETE and by Portuguese funds from FCT through the project PEst-OE/BIA/UI4050/2014. By the Spanish Minister of Economy and Competitiveness (MAT2012-38043-C02-01) and Junta de Castilla y León-JCyL (VA152A12-2 and VA155A12-2), Spain. AC, VS and RM, acknowledge FCT for SFRH/BD/75882/2011, SFRH/BPD/63148/2009 and SFRH-BPD/86470/2012 grants, respectively. The authors also thank support from the COST Action MP1206 “Electrospun Nano-fibres for bio inspired composite materials and innovative industrial applications”.info:eu-repo/semantics/publishedVersio

Electrospun silk-elastin fibres functionalized with silver nanoparticles as antibacterial wound dressings

[Excerpt] Silk-elastin-like proteins (SELPs) are a class of bioinspired, genetically engineered block copolymers, composed of silk and elastin repeating units. As base materials for biomedical purposes, SELP nanofibre mats demonstrate potential to be applied as wound dressing materials [1]. The increasing antimicrobial resistance associated with the excessive and inappropriate use of antibiotics demands the research for new pathogen-free healthcare polymeric materials with enhanced biological performance. [...]This work was supported by the strategic programme UID/BIA/04050/2013 (POCI-01- 0145-FEDER- 007569) through FCT I.P. and by ERDF through COMPETE2020 - POCI. The authors are grateful for funding from FCT (project “FunBioPlas” ERA-IB-2-6/0004/2014) and a fellowship to RM (SFRH-BPD/86470/2012). The authors also thank support from the COST Action MP1206

Synthetic Protein Biotechnology approaches for the creation of antimicrobial biopolymers

[Excerpt] The spread of antimicrobials resistant microorganisms has triggered the search for new ways to treat infections. 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. [...]This work was supported by FEDER through POFC – COMPETE by FCT through the project PEst-OE/BIA/UI4050/2014. By the Spanish Minister of Economy and Competitiveness (MAT2012-38043-C02-01) and Junta de Castilla y León-JCyL (VA152A12-2 and VA155A12-2), Spain. AC and RM, acknowledge FCT for SFRH/BD/75882/2011 and SFRH-BPD/86470/2012 grants, respectively

Antimicrobial Electrospun Membranes of Selp/Ag Composites

Silk-elastin-like proteins (SELPs) are a new class of bioinspired, biologically synthesized block copolymers, composed of silk and elastin repeating units. SELP electrospun fibre mats show potential for application as wound dressings for skin regeneration. In this work, antimicrobial nanofibrous mats were produced by electrospinning SELP solutions containing different concentrations of silver nitrate without addition of reducing agents. The SELP/Ag composite materials demonstrated antimicrobial activity against both Gram– and Gram+ bacteria. Furthermore, the SELP/Ag composite materials showed no cytotoxicity against normal human skin fibroblasts.This work was supported by FCT/MEC through Portuguese funds (PIDDAC) - PEst-OE/BIA/UI4050/2014, PEST-C/FIS/UI607/2011, Matepro - NORTE-07-0124-FEDER-000037. RM, AC and VS, acknowledge FCT for SFRH-BPD/86470/2012, SFRH/BD/75882/2011 and SFRH/BPD/63148/2009 grants, respectively. The authors also thank support from the COST Action MP1206 “Electrospun Nano-fibres for bio inspired composite materials and innovative industrial applications”

Genetically engineered silk-based composite biomaterials functionalized with fibronectin type-II that promote cell adhesion

[Excerpt] Recombinant protein-based polymers (rPBPs) are an emerging class of biopolymers inspired by Nature and produced by synthetic protein biotechnology approaches. Due to their exceptional physical-chemical and biological characteristics, as well as their ability to be customized for specific applications, rPBPs have been explored for the development of advanced biomaterials [1]. Within rPBPs, silk-like polymers (SLP) are being utilized in a range of studies in materials science [2]. [...]This work was supported by FCT Funded Project “Chimera” (PTDC/EBB-EBI/109093/2008), by FCT/MEC through Portuguese funds (PIDDAC) – PEst-OE/BIA/UI4050/2014, by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI). TC is thankful to the FCT for its support through Investigador FCT 2015. ARibeiro thanks FCT for the SFRH/BPD/98388/2013 grant. RMachado and AdaCosta acknowledge FCT for SFRH-BPD/86470/2012 and SFRH/BD/75882/2011 grants, respectively

Genetically engineered Silk-Elastin-Like Proteins as a versatile platform for the development of new biomaterials

[Excerpt] Throughout evolution nature created and refined proteins for a wide range of functions, working as structural components or as molecular motors. In this sense, the natural fibrous proteins represent the utmost case of function specialization and high performance materials. The remarkable mechanical properties of proteins like elastin or silk are founded on conservative blocks of amino acid sequences that propagate through the natural protein. These repetitive amino acid sequences are arranged in a way that creates flexible, rigid or tough domains, which are responsible for the physical and mechanical properties of the natural protein. Indeed, the recognition of the mechanics linking the nano- and micro-scale structure with the macromolecular assembly and organization, enabled molecular biologists to understand nature’s refined ways of creating high performance structural materials. Advances in synthetic protein biotechnology, emerging from the increase of knowledge in structural and molecular biology, combined with the use of recombinant DNA technology and biotechnology processes, made possible the advent of a new class of artificial biomacromolecules, the recombinant Protein-Based Polymers (rPBPs). This new class of protein-based materials, inspired in nature and with precisely controlled amino acid sequences, mimic the properties of their natural counterparts but can also combine in the same polypeptide chain the properties of two or more different proteins, creating copolymers with distinct properties from their native equivalents. Indeed, by recombinant DNA technology, it is possible to design and produce tailored synthetic genes, allowing for the creation of multifunctional complex PBPs with absolute control of its composition, structure and molecular weight. [...]This work is supported by the strategic programme UID/BIA/04050/2013 (POCI-01-0145-FEDER-007569) funded by national funds through the FCT I.P. and by the ERDF through COMPETE2020 – Programa Operacional Competitividade e Internacionalização (POCI) and by the project EcoAgriFood (NORTE-01-0145-FEDER-000009), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). It is also supported by FCT within the ERA-NET IB, project FunBioPlas with grant number ERA-IB-15-089 and FCT reference ERA-IB-2-6/0004/2014. AMPereira acknowledges DP_AEM and FCT for the PD/BD/113811/2015 grant

Advanced silk-based genetic polymers with improved cell adhesion properties

[Excerpt] Recombinant protein-based polymers (rPBPs) are an emerging class of genetic polymers inspired by Nature and produced by synthetic protein biotechnology approaches. Due to their exceptional physical-chemical and biological characteristics, as well as their ability to be customized for specific applications, rPBPs have been explored for the development of advanced biomaterials [1]. Most of the polymers used as biomaterials thus far have been chemically synthesized, originating random copolymers with diverse and uncontrolled distribution of molecular weight (MW) and composition. However, advances in recombinant DNA technology allow the biological synthesis of fine-tuned rPBPs with precise control of their composition, polymer size and structure [2]. Furthermore, with the development of recombinant protein engineering and biotechnology, it is now possible to design new bioactive rPBPs by combining active peptides/domains from different natural proteins in the same fusion protein. [...