Functional protein films as interfacial coadjuvants: A synergistic strategy to enhance antibiotic efficacy and suppress biofilms

Increasing market demand for sustainable, environmentally friendly edible film materials has called for the development of new customizable production methods utilizing emerging technologies such as 3D printing. We hereby report a new method to generate functional edible soy protein isolate films prepared from three types of soybeans (AR-R11-7999, MO-S17-17168, and MO-S17-19874R) using an innovative 3D printing technology. The protein contents in AR-R11-7999, MO-S17-17168, and MO-S17-19874R soybean meals and their corresponding protein isolates were 40.0, 39.1, and 39.9; and 84.5, 84.7, and 87.3 % (w/w, dry basis), respectively. Response surface methodology was used to maximize the tensile and puncture strength and minimize the thickness of the 3D-printed edible films using protein concentration, plasticizer concentration (glycerol), and drying time as the independent variables. The optimized film production conditions were determined as soy protein concentration: 8.91%, plasticizer concentration: 3.00%, and drying time: 3.98h with a desirability value of 0.7428. The optimized conditions were then successfully verified with the original soybean lot with a nonsignificant difference in physical properties. At the optimized conditions, the 3D-printed edible films using three soybean lots revealed: 0.108-0.114mm thickness; 14.79-16.07MPa tensile strength; 6.97-8.20 N puncture strength; 90.81-91.53, -1.89 to -1.31, and 14.85-17.25 were color parameters L*, a*, and b*, respectively; 1.22-1.36g/cm3 density; and 104.4-105.7% elongation at break ratio (%). PRACTICAL APPLICATION: Edible soy protein films produced by an extrusion-based 3D printing approach are highly customizable and precise, and could be produced at an industrial scale. This newly produced environment-friendly soy protein-based edible film can serve as an alternate packaging to synthetic plastics and reduce the environmental landfill problem while adding value to soybean produced in the mid-south United States.

The ability to add bioactivities, such as cell signaling or ligand recognition, to biomaterials has generated the potential to include multiple bioactivities into a single material. In some cases, it is desirable to localize these activities to different areas of the biomaterial, creating functional patterns. While photolithography and 3D printing have been effective techniques for patterning functions in many materials, patterning remains a challenge in materials composed of protein, in part due to how these materials are artificially assembled. Protein fibers are often produced from protein films that co-acervate at the air-water interface. This chapter describes methods to leverage this coacervation process to pattern materials, using the Drosophila melanogaster Hox protein Ultrabithorax (Ubx) as a model self-assembling protein. Through gene fusion, Ubx and a functional protein are produced as a single polypeptide, capable of both forming materials and performing the activity of interest. This functionality is retained in the final materials. In this chapter, we describe how to use multiple Ubx fusion proteins to not only imbue the final materials with multiple functions, but also to create macroscale patterns of the appended proteins in fibrous protein-based materials. These patterned materials include striped fibers, bifunctional-faced fibers, gradient fibers, and core-shell fibers.

Probing fundamental film parameters of immobilized enzymes—Towards enhanced biosensor performance. Part II—Electroanalytical estimation of immobilized enzyme performance

Antimicrobial effect of functional protein films incorporating garlic oil (GO), potassium sorbate (PS) and nisin (N) at various concentrations were discussed. This activity was tested against food pathogenic bacteria namely Escherichia coli (E.coli) and Staphylococcus aureus (S.aureus).Mechanical and physical properties were characterized. In the range of antimicrobial agents concentration studied, tensile strength (TS) and elongation at break (E) of functional protein films were changed by incorporating GO,PS and N. And the WVP value of functional protein films decreased as antimicrobial agents added. GO incorporated into protein films had no effect on E.coli, but incorporation of GO at 300μl had antimicrobial activity against S.aureus. Protein films incorporated with PS showed antimicrobial activity against S.aureus, but there was no effect on E.coli. Incorporation of N at the lowest level of 25,000 IU had antimicrobial activity against both E.coli and S.aureus.

Heat-denatured and alcalase-hydrolyzed protein films/coatings containing marjoram essential oil and thyme extract

A superhydrophobic and self-cleaning photoluminescent protein film with high weatherability

Biosynthesized gold nanoparticles from Proteus mirabilis: A synergistic strategy against multidrug-resistant Gram-negative bacteria.

Sustainable and functional tandem repeat protein fibers and films

Sustainable and functional tandem repeat protein fibers and films

Sustainable and functional tandem repeat protein fibers and films

The impact of protein and glycerol concentration on the optical, mechanical, and moisture sorption properties of whey protein isolate edible films was determined. The increasing of protein concentration leads to decreasing opacity and moisture adsorption of films but improving its tensile strength and elongation at break. Color parameters for all films remain constant and were not dependent on the film composition. Films with increasing glycerol concentration showed weakened mechanical resistance and higher moisture adsorption rates. Moisture sorption isotherms data were well described by GAB, Peleg, and Lewicki equations. These findings can be used to better design applications of whey protein films containing plasticizers, in order to optimize film formulation in a rational manner considering their practical use as protective coatings or edible packaging.Practical applicationsThe increased interest in the development of edible films for food packaging applications has been raised to minimize the use of synthetic petroleum‐based polymers. Moisture sensitivity of films showed the capacity of such materials to modify their physical properties which are influenced by different relative humidity or temperature conditions. The results presented in this study, concerning optical, mechanical, and sorption properties of whey protein‐based edible films, would provide the scientific basis for developing functional protein films and their application for food products including nuts, seeds, fruits, or vegetables.

Generating Novel Materials Using the Intrinsically Disordered Protein Ubx.

RuBisCO-based protein and film from Rumex patience L.: Effects of green extraction and NADES plasticization on structural and functional properties.

Immobilization of azurin with retention of its native electrochemical properties at alkylsilane self-assembled monolayer modified indium tin oxide

ABSTRACTAccurate and timely functional genome annotation is essential for translating basic pathogen research into clinically impactful advances. Here, through literature curation and structure-function inference, we systematically update the functional genome annotation of Mycobacterium tuberculosis virulent type strain H37Rv. First, we systematically curated annotations for 589 genes from 662 publications, including 282 gene products absent from leading databases. Second, we modeled 1,711 underannotated proteins and developed a semiautomated pipeline that captured shared function between 400 protein models and structural matches of known function on Protein Data Bank, including drug efflux proteins, metabolic enzymes, and virulence factors. In aggregate, these structure- and literature-derived annotations update 940/1,725 underannotated H37Rv genes and generate hundreds of functional hypotheses. Retrospectively applying the annotation to a recent whole-genome transposon mutant screen provided missing function for 48% (13/27) of underannotated genes altering antibiotic efficacy and 33% (23/69) required for persistence during mouse tuberculosis (TB) infection. Prospective application of the protein models enabled us to functionally interpret novel laboratory generated pyrazinamide (PZA)-resistant mutants of unknown function, which implicated the emerging coenzyme A depletion model of PZA action in the mutants’ PZA resistance. Our findings demonstrate the functional insight gained by integrating structural modeling and systematic literature curation, even for widely studied microorganisms. Functional annotations and protein structure models are available at https://tuberculosis.sdsu.edu/H37Rv in human- and machine-readable formats.IMPORTANCEMycobacterium tuberculosis, the primary causative agent of tuberculosis, kills more humans than any other infectious bacterium. Yet 40% of its genome is functionally uncharacterized, leaving much about the genetic basis of its resistance to antibiotics, capacity to withstand host immunity, and basic metabolism yet undiscovered. Irregular literature curation for functional annotation contributes to this gap. We systematically curated functions from literature and structural similarity for over half of poorly characterized genes, expanding the functionally annotated Mycobacterium tuberculosis proteome. Applying this updated annotation to recent in vivo functional screens added functional information to dozens of clinically pertinent proteins described as having unknown function. Integrating the annotations with a prospective functional screen identified new mutants resistant to a first-line TB drug, supporting an emerging hypothesis for its mode of action. These improvements in functional interpretation of clinically informative studies underscore the translational value of this functional knowledge. Structure-derived annotations identify hundreds of high-confidence candidates for mechanisms of antibiotic resistance, virulence factors, and basic metabolism and other functions key in clinical and basic tuberculosis research. More broadly, they provide a systematic framework for improving prokaryotic reference annotations.