Novel Bovine Plasma Protein Film Reinforced with Nanofibrillated Cellulose Fiber as Edible Food Packaging Material

Proteins, such as those in blood from slaughterhouses, are a good option for developing edible films. However, films made exclusively from proteins have low strength and high water solubility, which makes them difficult to use in the food industry. The use of cellulosic material, such as nanofibrillated cellulose (NFC), can improve the properties of these films. In the present work, bovine plasma was acidified and treated with ethanol to precipitate its proteins, and these proteins were used to prepare films reinforced with several concentrations of NFC. In addition, control films prepared with untreated bovine plasma and reinforced with NFC were prepared as well. These new edible films were characterized according to their mechanical properties, water vapor permeability, light transmittance, and microstructure. Furthermore, the film with the best properties was selected to be additivated with nisin to test its antimicrobial properties by wrapping meat previously contaminated with Staphylococcus aureus. In this sense, films prepared with the extracted proteins showed better properties than the films prepared with untreated plasma. In addition, the results showed that the reinforcement of the films with a 10% (w/w) of NFC decreased their water solubility and improved their puncture strength and water vapor barrier properties. Finally, the addition of nisin to the films prepared with extracted protein from bovine plasma and NFC gave them antimicrobial properties against S. aureus.

Fabrication of flexible conductive silk fibroin/polythiophene membrane and its properties

Silk fibroin (SF) film is an insulating material, which can be combined with polythiophene derivatives with electrical conductivity to obtain a flexible conductive material. In this work, poly(3,4-ethylenedioxythiophene) (PEDOT) was used to graft a silk protein film. The hydroxyl radical is formed by activation and oxidation of the silk protein film polymerized with the PEDOT radical formed by oxidation of 3,4-ethylenedioxythiophene to obtain a conductive silk film. The SF/PEDOT film, when tested, showed excellent electrical conductivity with resistance up to 63 Ω·cm−2, good flexibility, mechanical properties, fastness, and biocompatibility.

Surface-enhanced Raman scattering of CoV-SARS-2 viral proteins in a strong coupling regime

Fast, sensitive, high-throughput detection of coronavirus antigens at physiologically relevant levels is essential for population screening that could prevent epidemics such as the current COVID-19 global pandemic. Optical methods based on surface-enhanced Raman scattering (SERS) spectroscopy are promising for this purpose because they ensure quick detection of even single biological molecules in a sample. For achieving such a high sensitivity, it is crucial to design SERS-active systems concentrating incident radiation into small sample volumes. Here, metal-dielectric cavities have been obtained through interaction of protein sulfhydryl groups with a SERS-active silver surface. The concentration of light in these cavities allows the differential detection of spike glycoprotein and nucleocapsid protein of SARS-COV-2, which are its key antigens, at physiologically relevant concentrations. The cavity Q-factor can be increased by additionally covering the dielectric protein film with a silver shell to form an ultrathin cavity, which provides an at least tenfold enhancement of the detection signal. The results could be used to design high-throughput systems for specific and sensitive detection of viral antigens and quick diagnosis of viral infections.

SIMPLE ASSAY FOR ADSORPTION OF PROTEINS TO THE AIR-WATER INTERFACE

A rapid assay is described, based upon the Marangoni effect, which detects the formation of a denatured-protein film at the air-water interface (AWI) of aqueous samples. This assay requires no more than a 20 microliter aliquot of sample, at a protein concentration of no more than 1 mg/ml, and it can be performed with any buffer that is used to prepare grids for electron cryo-microscopy (cryo-EM). In addition, this assay provides an easy way to estimate the rate at which a given protein forms such a film at the AWI. Use of this assay is suggested as a way to pre-screen the effect of various additives and chemical modifications that one might use to optimize the preparation of grids, although the final proof of optimization still requires further screening of grids in the electron microscope. In those cases when the assay establishes that a given protein does form a sacrificial, denatured-protein monolayer, it is suggested that subsequent optimization strategies might focus on discovering how to improve the adsorption of native proteins onto that monolayer, rather than to prevent its formation. A second alternative might be to bind such proteins to the surface of rationally designed affinity grids, in order to prevent their diffusion to, and unwanted interaction with, the AWI.

The Inhibition Study of Cytochrome bd Oxidase Using the Enzyme-Based Electrochemical Sensor

Membrane proteins that participate in multiple vital functions of every living organism such as transport, signaling and respiration, provide 80 to 90% of the relevant targets for the pharmaceutical industries. The family of cytochrome bd oxidase enzymes is of great interest for the development of future antibiotics as they are found only in the respiratory chain of the prokaryotes and they are believed to be involved in bacterial adaptability mechanisms. They catalyze the reduction of molecular oxygen in water and oxidation of quinols and contribute to the proton motive force required for ATP synthesis. Due to their hydrophobic nature, membrane proteins are more difficult to handle than soluble proteins. Protein film voltammetry is a very convenient technique, because it allows for working at a very low concentration and for optimizing the electrode surface to the nature of the enzyme. Here, we have developed a biosensor for the study of terminal oxidases based on their immobilization on gold nanoparticles modified with a self-assembled monolayer of thiols. The stability of the protein films can be optimized by varying the nature of thiols and amount of lipids. This enzyme-based electrochemical sensor was successfully used for the inhibition screening of a target-focused library of 34 compounds which belong to the families of quinones, naphthoquinones, phenols, quinolones, coumarins and flavonoids against cytochrome bd oxidase. Moreover, the developed device was applied for the study of the catalytic reaction of the enzyme with small gaseous signaling molecules.

Development and Characterization of Novel Composite Films Based on Soy Protein Isolate and Oilseed Flours

The possibility of using oilseed flours as a waste source for film-forming materials with a combination of soy protein isolate in preparation of edible films was evaluated. Physical, mechanical and barrier properties were determined as a function of the oilseed type: hemp, evening primrose, flax, pumpkin, sesame and sunflower. It was observed that the addition of oilseed flours increased the refraction and thus the opacity of the obtained films from 1.27 to 9.57 A mm−1. Depending on the type of flours used, the edible films took on various colors. Lightness (L*) was lowest for the evening primrose film (L* = 34.91) and highest for the soy protein film (L* = 91.84). Parameter a* was lowest for the sunflower film (a* = −5.13) and highest for the flax film (a* = 13.62). Edible films made of pumpkin seed flour had the highest value of the b* color parameter (b* = 34.40), while films made of evening primrose flour had the lowest value (b* = 1.35). All analyzed films had relatively low mechanical resistance, with tensile strength from 0.60 to 3.09 MPa. Films made of flour containing the highest amount of protein, pumpkin and sesame, had the highest water vapor permeability, 2.41 and 2.70 × 10−9 g·m−1 s−1 Pa−1, respectively. All the edible films obtained had high water swelling values from 131.10 to 362.16%, and the microstructure of the films changed after adding the flour, from homogeneous and smooth to rough. All blended soy protein isolate–oilseed flour films showed lower thermal stability which was better observed at the first and second stages of thermogravimetric analysis when degradation occurred at lower temperatures. The oilseed flours blended with soy protein isolate show the possibility of using them in the development of biodegradable films which can find practical application in the food industry.

A Multiple-Technique Approach to Inferring Bio-electrochemical Reaction Parameters

Uncovering the secrets of the biological Faradaic reactions, essential to the understanding of complex metalloenzymes, requires an information recovery process that is robust, rapid and replicable. This paper is a description of the workflow we have developed over the course of inferring chemical reaction parameters for a simple protein system, a bacterial cytochrome domain from \textit{Cellvibrio japonicus}. This was a challenging task, as the signal-to-noise ratio in such protein-film voltammetry experiments is significantly lowered relative to the voltammetric data generated by simple chemicals. We have overcome these challenges by using a multiple-technique approach, which compensates for the difficulties inherent to analysis of the individual voltammetry experiments. We have shown that the parameters inferred are robust across multiple experiments performed for different preperations of the protein. This is an important proof-of-concept result for analysis of more complex metalloenzymes, which incorporate catalytic processes and multiple internal electron-transfer sites. <br>