RESUMEN
The immobilization of fluorescent proteins is a key technology enabling to fabricate a new generation of photoactive materials with potential technological applications. Herein we have exploited superfolder green (sGFP) and red (RFP) fluorescent proteins expressed with different polypeptide tags. We fused these fluorescent proteins to His-tags to immobilize them on graphene 3D hydrogels, and Cys-tags to immobilize them on porous microparticles activated with either epoxy or disulfide groups and with Lys-tags to immobilize them on upconverting nanoparticles functionalized with carboxylic groups. Genetically programming sGFP and RFP with Cys-tag and His-tag, respectively, allowed tuning the protein spatial organization either across the porous structure of two microbeads with different functional groups (agarose-based materials activated with metal chelates and epoxy-methacrylate materials) or across the surface of a single microbead functionalized with both metal-chelates and disulfide groups. By using different polypeptide tags, we can control the attachment chemistry but also the localization of the fluorescent proteins across the material surfaces. The resulting photoactive material formed by His-RFP immobilized on graphene hydrogels has been tested as pH indicator to measure pH changes in the alkaline region, although the immobilized fluorescent protein exhibited a narrower dynamic range to measure pH than the soluble fluorescent protein. Likewise, the immobilization of Lys-sGFP on alginate-coated upconverting nanoparticles enabled the infrared excitation of the fluorescent protein to be used as a green light emitter. These novel photoactive biomaterials open new avenues for innovative technological developments towards the fabrication of biosensors and photonic devices.
Asunto(s)
Grafito/química , Proteínas Fluorescentes Verdes/química , Hidrogeles/química , Proteínas Inmovilizadas/química , Proteínas Luminiscentes/química , Proteínas Recombinantes de Fusión/química , Alginatos/química , Técnicas Biosensibles , Expresión Génica , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Histidina/química , Histidina/genética , Histidina/metabolismo , Concentración de Iones de Hidrógeno , Proteínas Inmovilizadas/genética , Proteínas Inmovilizadas/metabolismo , Luz , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Metacrilatos/química , Nanopartículas/química , Oligopéptidos/química , Oligopéptidos/genética , Oligopéptidos/metabolismo , Procesos Fotoquímicos , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Sefarosa/química , Proteína Fluorescente RojaRESUMEN
Encapsulation of active agents, such as vitamins and antioxidants, is one of the possibilities that allow their incorporation in beverages, food, or in pharmaceutical products. Simultaneously, encapsulation protects these active agents from oxidation, producing more stable active compounds. Formation of nanodroplets by spontaneously formed microemulsion (ME) offers, on one hand, a low-energy technology of encapsulation and, on the other hand, because of a small size of the droplets, it assures long-term stability even in harsher environments. In this study, oil-in-water MEs allowed the low-energy encapsulation of α-tocopherol (αToc) into an aqueous medium with the aid of fully food-grade ingredients, using isoamyl acetate as the dispersed oil phase, which was selected between three different types of oils. Both cosurfactant-free and cosurfactant-holder ME systems were formulated, in which Tween 20 and glycerol were employed as the surfactant and the cosurfactant, respectively. The ME monophasic area was determined through the construction of pseudoternary phase diagrams. The encapsulated αToc within 10-20 nm nanocapsules showed radical scavenging activity dependent on the encapsulated amount of αToc, as it was demonstrated by electron paramagnetic resonance spectroscopy. The radical scavenging activity slightly increased within the time investigated, indicating a slow release of the active compound from the nanodroplets, which is a promising result for their application, especially in pharmaceuticals.
RESUMEN
A waterborne pressure-sensitive adhesive (PSA) that shows high adhesive performance and easy debondability on demand without leaving residues on the substrate (adhesive failure) has been developed. A key component of the PSA is a semicrystalline phase that is beneficial for the adhesive properties and that becomes fluid when heated above the melting temperature. Migration of this liquid-like polymer to the substrate-adhesive interface and hardening upon cooling results in a hard non-tacky interface that facilitates debonding. The effect of the particle morphology on the debonding ability is discussed.
