The number of methodologies for the immobilization of enzymes using polymeric supports is continuously growing due to the developments in the fields of biotechnology, polymer chemistry, and nanotechnology in the last years. Despite being excellent catalysts, enzymes are very sensitive molecules and can undergo denaturation beyond their natural environment. For overcoming this issue, polymer chemistry offers a wealth of opportunities for the successful combination of enzymes with versatile natural or synthetic polymers. The fabrication of functional, stable, and robust biocatalytic hybrid materials (nanoparticles, capsules, hydrogels, or films) has been proven advantageous for several applications such as biomedicine, organic synthesis, biosensing, and bioremediation. In this review, supported with recent examples of enzyme-protein hybrids, we provide an overview of the methods used to combine both macromolecules, as well as the future directions and the main challenges that are currently being tackled in this field.

Bio-hybrid light-emitting diodes (Bio-HLEDs) based on color down-converting filters with fluorescent proteins (FPs) have achieved moderate efficiencies (50 lm/W) and stabilities (300 h) due to both thermal-and photo-degradation. Here, we present a significant enhancement in efficiency (~130 lm/W) and stability (>150 days) using a zero-thermalquenching bio-phosphor design. This is achieved shielding the FP surface with a hydrophilic polymer allowing their homogenous integration into the network of a light-guiding and hydrophobic host polymer. We rationalize how the control of the mechanical and optical features of this bio-phosphor is paramount towards highly stable and efficient Bio-HLEDs, regardless of the operation conditions. This is validated by the relationships between the stiffness of the FP-polymer phosphor and the maximum temperature reached under device operation as well as the transmittance of the filters and device efficiency.

A simple approach for the fabrication of functional nanopatterned protein materials using protein engineering and soft-nanolithography and its implementation in optical devices based on distributed feedback (DFB) laser phenomena.

There is a current need to fabricate new biobased functional materials. Bottom‐up approaches to assemble simple molecular units have shown promise for biomaterial fabrication due to their tunability and versatility for the incorporation of functionalities. Herein, the fabrication of catalytic protein thin films by the entrapment of catalase into protein films composed of a scaffolding protein is demonstrated. Extensive structural and functional characterization of the films provide evidence of the structural integrity, order, stability, catalytic activity, and reusability of the biocatalytic materials. Finally, these functional biomaterials are coupled with piezoelectric disks to fabricate a second generation of bio‐inorganic generators. These devices are capable of producing electricity from renewable fuels through catalase‐driven gas production that mechanically stimulates the piezoelectric material.

This work presents a new family of bio-hybrid light-emitting diodes (Bio-HLEDs) using all-bio color down-converting coatings that combine silk fibroin (SF) as a packaging matrix and fluorescent proteins (FPs) as emitters.

Nevertheless, the utilization of biomolecules, i.e., proteins, to impart functionality to inorganic and/or organic materials and afford highly efficient functional devices presents a number of challenges in the research of functional biomaterials. [4][5][6] The main limitations predominantly arise from intermolecular aggregation, surfaceinduced denaturation, steric hindrance of active sites, and lack of dynamical freedom imposed by solid state. [7][8][9] The deposition of continuous protein thin films seems to be a good strategy that fulfill those needs. [10,11] With particular emphasis on biocatalytic coatings, the fabrication method should guarantee high enzyme loads, low substrate/product flow transport limitations, and improve the lifetime and stability of the biomolecule. [12] Currently, reported methods for the fabrication of functional biofilms are based on the utilization of a relatively limited range of naturally self-assembling proteins, layer-by-layer deposition approaches, and the adsorption of the proteins to amphiphilic copolymers. [13][14][15][16][17] However, these approaches usually require of the covalent crosslinking of the components in order to avoid the disaggregation of the film in water and at broad range of pH. [18] Yet, the uncontrolled covalent crosslinking might be especially damaging in the formation of functional protein films. The protein's amino acids can be altered and severe substrate diffusion issues might be caused within the film, resulting in impaired biomaterials. [19] Therefore, an alternative sequence-independent methodology that allows the fabrication of functional protein films would vastly expand the toolkit for creating biomaterials.In this regard, the bioinspired self-assembly of hierarchically structured peptide or protein films is an attractive approach. [20][21][22][23] In nature, the metal-driven crosslinking of specific peptidic building blocks leads to complex hierarchical structures across many lengths, as it happens in mussel byssus or worm jaws. [24][25][26] Furthermore, metal-directed protein selfassembly (MDPSA) methodology is inspired by the affinity of distinct residue side chains such as histidines, cysteines, lysines, and asparagines toward metallic cations (mainly Ni, Cu, Co, and Zn). MDPSA allocates such key residues on the surface of the protein as anchoring points. [27][28][29] Hence, metal ions are used as inorganic bridges that not only guide the assembly of the proteins into hierarchical architectures, but also might The deposition of protein thin films on (in)organic surfaces is a key approach to incorporate new functionalities into these materials for a broad number of applications. However, most of the current methods used for the controlled assembly of such biomolecules and eventual film formation are limiting since entail either the chemical modification of the proteins, which leads sometimes to impaired materials, or the sequential layer-by-layer deposition of charged macromolecules. In this work, a facile bioinspired method for the versati...

In nature, assembled protein structures offer the most complex functional structures. The understanding of the mechanisms ruling protein-protein interactions opens the door to manipulate protein assemblies in a rational way. Proteins are versatile scaffolds with great potential as tools in nanotechnology and biomedicine because of their chemical, structural, and functional versatility. Currently, bottom-up self-assembly based on biomolecular interactions of small and well-defined components, is an attractive approach to biomolecular engineering and biomaterial design. Specifically, repeat proteins are simplified systems for this purpose. In this work, we provide an overview of fundamental concepts of the design of new protein interfaces. We describe an experimental approach to form higher order architectures by a bottom-up assembly of repeated building blocks. For this purpose, we use designed consensus tetratricopeptide repeat proteins (CTPRs). CTPR arrays contain multiple identical repeats that interact through a single inter-repeat interface to form elongated superhelices. Introducing a novel interface along the CTPR superhelix allows two CTPR molecules to assemble into protein nanotubes. We apply three approaches to form protein nanotubes: electrostatic interactions, hydrophobic interactions, and π-π interactions. We isolate and characterize the stability and shape of the formed dimers and analyze the nanotube formation considering the energy of the interaction and the structure in the three different models. These studies provide insights into the design of novel protein interfaces for the control of the assembly into more complex structures, which will open the door to the rational design of nanostructures and ordered materials for many potential applications in nanotechnology.

Herein, we report a multifunctional chemoenzymatic nanoreactor (NanoNOx) for the glucosecontrolled regeneration of natural and artificial nicotinamide cofactors. NanoNOx are built of glucose oxidasepolymer hybrids that assemble in the presence of an organometallic catalyst: hemin. The design of the hybrid is optimized to increase the effectiveness and the directional channeling at low substrate concentration. Importantly, NanoNOx can be reutilized without affecting the catalytic properties, can show high stability in the presence of organic solvents, and can effectively oxidize assorted natural and artificial enzyme cofactors. Finally, the hybrid was successfully coupled with NADHdependent dehydrogenases in one-pot reactions, using a strategy based on the sequential injection of a fuel, namely, glucose. Hence, this study describes the first example of a hybrid chemoenzymatic nanomaterial able to efficiently mimic NOx enzymes in cooperative onepot cascade reactions.