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Projeto de investigação
Protein materials for gas sensing
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A scalable method to purify reflectins from inclusion bodies
Publication . Lychko, Iana; Soares, Cátia Lopes; Dias, Ana Margarida Gonçalves Carvalho; Roque, Ana Cecília Afonso; UCIBIO - Applied Molecular Biosciences Unit; DQ - Departamento de Química; Elsevier
Structural proteins are an attractive inspiration for functional biobased materials. In nature, cephalopods skin colour modulation is related to the dynamic self-assembly of a family of structural proteins known as reflectins. To fully reach their potential as engineered bio-based materials, reflectins need to be produced by biotechnological means. One of the challenges is associated with establishing and optimizing reflectin purification processes to achieve the highest yield and productivity. Here, we studied purification strategies for two reflectin sequences from different organisms which were recombinantly expressed in a bacterial host at laboratory scale. Reflectins purification was then assessed by two chromatographic and one non-chromatographic methods. Methods were compared considering final purity and yield, productivity, cost and sustainability. The non-chromatographic method based on inclusion bodies washing presented the most promising results (protein purity > 90% and purification yields up to 88%). Our results contribute to define bioprocessing strategies to address the vision of biodegradable and sustainable protein-based materials.
Optimizing the production and processing of Reflectin proteins and derived materials
Publication . Lychko, Iana; Roque, Ana; Dias, Ana
Reflectins are structural proteins that play a vital role in cephalopods' camouflage being found in specialized reflecting cells. They have inspired the development of several bio-photonic prototype systems with potential applications in camouflage, sensing, and advanced optical technologies. However, to reach their full potential, the biotechnological production of reflectins must be optimized and a deep understanding of theoretical and experimental aspects related to reflectins condensation, assembly and processing requires attention. This thesis aimed to address this challenge, by presenting a comprehensive investigation of reflectins purification to maximize production yields; followed by the in vitro study of conditions that affect reflectins self-assembly and condensation, and finally assessment of distinct reflectins processing methods into materials.
Solvent modulation in peptide sub-microfibers obtained by solution blow spinning
Publication . Dias, Ana Margarida Gonçalves Carvalho; Cena, Cícero; Lutz-Bueno, Viviane; Mezzenga, Raffaele; Marques, Ana; Ferreira, Isabel; Roque, Ana Cecília Afonso; DQ - Departamento de Química; UCIBIO - Applied Molecular Biosciences Unit; CENIMAT-i3N - Centro de Investigação de Materiais (Lab. Associado I3N); DCM - Departamento de Ciência dos Materiais; Frontiers Media
Peptides possess high chemical diversity at the amino acid sequence level, which further translates into versatile functions. Peptides with self-assembling properties can be processed into diverse formats giving rise to bio-based materials. Peptide-based spun fibers are an interesting format due to high surface-area and versatility, though the field is still in its infancy due to the challenges in applying the synthetic polymer spinning processes to protein fibers to peptides. In this work we show the use of solution blow-spinning to produce peptide fibers. Peptide fiber formation was assisted by the polymer poly (vinyl pyrrolidone) (PVP) in two solvent conditions. Peptide miscibility and further self-assembling propensity in the solvents played a major role in fiber formation. When employing acetic acid as solvent, peptide fibers (0.5 μm) are formed around PVP fibers (0.75 μm), whereas in isopropanol only one type of fibers are formed, consisting of mixed peptide and PVP (1 μm). This report highlights solvent modulation as a mean to obtain different peptide sub-microfibers via a single injection nozzle in solution blow spinning. We anticipate this strategy to be applied to other small peptides with self-assembly propensity to obtain multi-functional proteinaceous fibers.
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Entidade financiadora
Fundação para a Ciência e a Tecnologia
Programa de financiamento
OE
Número da atribuição
SFRH/BD/147388/2019