Quantification of diffusion coefficients of commonly used high-intensity sweeteners through mucin.

Low- and no-calorie sweeteners reduce the amount of carbohydrates in foods and beverages. However, concerns about taste perception surrounding the role of non-nutritive sweeteners in the oral cavity remain unanswered. One of the parameters that influences taste perception is the diffusion coefficient of the sweetener molecules inside the mucin layer lining the mouth. This study investigated the impact of diffusion coefficients of common high-intensity sweeteners on taste perception focusing on the sweeteners' diffusion through mucin. Transwell Permeable Support well plates were used to measure diffusion coefficients of samples that were collected at specific intervals to estimate the coefficients based on concentration measurements. The diffusion coefficients of acesulfame-K, aspartame, rebaudioside M, sucralose, and sucrose with and without NaCl were compared. We found that different sweeteners show different diffusion behavior through mucin and that the presence of salt enhances the diffusion. These findings contribute insights into the diffusion of high-intensity sweeteners, offer a way to evaluate diffusion coefficients in real-time, and inform the development of products with improved taste profiles.

Substitution and Orientation Effects on the Crystallinity and PFAS Adsorption of Olefin-Linked 2D COFs.

Solid phase adsorbents with high removal affinity for per- and polyfluoroalkyl substances (PFAS) in aqueous environments are sought. We report the synthesis and investigation of COF-I, a new covalent organic framework (COF) with a good affinity for PFAS adsorption. COF-I was synthesized by the condensation reaction between 2,4,6-trimethyl-1,3,5-triazine and 2,3-dimethoxyterephthaldehyde and fully characterized. In addition to the high crystallinity and surface area, COF-I showed high hydrolytic and thermal stability. Further, we converted its hydrophobic surface to a hydrophilic surface by converting the ortho-methoxy groups to hydroxyl derivatives and produced a new hydrophilic olefin-linked two-dimensional (2D) COF. We experimentally measured the crystallinity of both COFs by X-ray diffraction and used atomistic simulations coupled with cross-polarization/magic angle spinning solid-state nuclear magnetic resonance (CP/MAS ssNMR) to determine the relative amounts of AA-stacking and AB-stacking present. COF-I, with its hydrophobic surface and methoxy groups in the ortho positions, showed the best PFAS adsorption. COF-I reduced the concentration of perfluorooctanoic acid from 20 to 0.069 µg L-1 and to 0.052 µg L-1 for perfluorooctanesulfonic acid. These amounts are lower than the U.S. Environmental Protection Agency advisory level (0.070 µg L-1). High efficiency, fast kinetic adsorption, and reusability of COF-I are advantages of COF-I for PFAS removal from water.

Food LEGO: Building hollow cage and sheet superstructures from starch.

The idea of building large structures from small building blocks has had a long history in the human imagination, from the beautifully intricate shells assembled from silica by unicellular algae to the Egyptian pyramids built from stone. Carrying this idea into the food industry has important implications. Here, we introduce a Pickering emulsion platform for building superstructures like hollow cages and sheets using starch granules as building blocks. In food, these superstructures occupy up to six times more space than their constituent parts, thereby delivering a viscosity greater by an order of magnitude than unstructured starch. To achieve this higher viscosity, they use an alternative superstructure mechanism as opposed to the classic swelling mechanism of individual particles. These super-thickeners may reduce calories, cut production costs, and stretch the global food supply, demonstrating how we can design the future by playing with our food.

Improving the thermal stability of phytase using core-shell hydrogel beads.

A core-shell hydrogel bead system was designed to maintain the catalytic activity of phytase and protect its enzymatic functionality from heat treatment. The designed structure consists of a chitosan-phytase complex core and an alginate-carrageenan hydrogel shell. The core-shell hydrogel was optimized to improve phytase encapsulation efficiency and increase the thermal stability of the encapsulated phytase. After heat treatment, encapsulated phytase retained âˆ¼ 70 % of its catalytic activity and the same secondary structure of free phytase. Fourier transform infrared spectroscopy indicated strong intermolecular interactions between chitosan and phytase in the core, but little interaction between the core and the alginate and κ-carrageenan shell, this supports the structural and functional stability of the phytase. Differential scanning calorimetry confirmed that the designed core-shell structure had a higher melting point. Encapsulating phytase in a core-shell hydrogel bead can enhance the thermal stability of phytase, which broadens the potential applications for phytase delivery.

Adaptive Laboratory Evolution of Probiotics toward Oxidative Stress Using a Microfluidic-Based Platform.

Adaptive laboratory evolution (ALE) can be used to make bacteria less susceptible to oxidative stress. An alternative to large batch scale ALE cultures is to use microfluidic platforms, which are often more economical and more efficient. Microfluidic ALE platforms have shown promise, but many have suffered from subpar cell passaging mechanisms and poor spatial definition. A new approach is presented using a microfluidic Evolution on a Chip (EVoc) design which progressively drives microbial cells from areas of lower H2 O2 concentration to areas of higher concentration. Prolonged exposure, up to 72 h, revealed the survival of adaptive strains of Lacticaseibacillus rhamnosus GG, a beneficial probiotic often included in food products. After performing ALE on this microfluidic platform, the bacteria persisted under high H2 O2 concentrations in repeated trials. After two progressive exposures, the ability of L. rhamnosus to grow in the presence of H2 O2 increased from 1 mm H2 O2 after a lag time of 31 h to 1 mm after 21 h, 2 mm after 28 h, and 3 mm after 42 h. The adaptive strains have different morphology, and gene expression compared to wild type, and genome sequencing revealed a potentially meaningful single nucleotide mutation in the protein omega-amidase.

Faster sperm selected by rheotaxis leads to superior early embryonic development in vitro.

To understand the impact of sperm speed as they swim against the flow on fertilization rates, we created conditions similar to the female reproductive tract (FRT) on a microfluidic platform for sperm selection. Selected sperm were evaluated based on early development of fertilized embryos. Bovine and human spermatozoa were selected at various fluid flow rates within the device. We found that the speed of bovine spermatozoa increases as the flow rate increases and that the amount of DNA fragmentation index is lowered by increasing the flow rate. Bovine spermatozoa selected by our platform at low (150 µL h-1, shear rate 3 s-1), medium (250 µL h-1, shear rate 5 s-1), and high flow rates (350 µL h-1, shear rate 7 s-1) were used for fertilization and compared to sperm sorted by centrifugation. The samples collected at the highest flow rate resulted in the formation of 23% more blastocysts compared to the control. While selecting for higher quality sperm by increasing the flow rate does result in lower sperm yield, quality improvement and yield may be balanced by better embryonic development.

Tetrazine-Linked Covalent Organic Frameworks With Acid Sensing and Photocatalytic Activity.

The first synthesis and comprehensive characterization of two vinyl tetrazine-linked covalent organic frameworks (COF), TA-COF-1 and TA-COF-2, are reported. These materials exhibit high crystallinity and high specific surface areas of 1323 and 1114 m2 g-1. The COFs demonstrate favorable band positions and narrow band gaps suitable for light-driven applications. These advantages enable TA-COFs to act as reusable metal-free photocatalysts in the arylboronic acids oxidation and light-induced coupling of benzylamines. In addition, these TA-COFs show acid sensing capabilities, exhibiting visible and reversible color changes upon exposure to HCl solution, HCl vapor, and NH3 vapor. Further, the TA-COFs outperform a wide range of previously reported COF photocathodes. The tetrazine linker in the COF skeleton represents a significant advancement in the field of COF synthesis, enhancing the separation efficiency of charge carriers during the photoreaction and contributing to their photocathodic properties. TA-COFs can also degrade 5-nitro-1,2,4-triazol-3-one (NTO), an insensitive explosive present in industrial wastewater, in 20 min in a sunlight-driven photocatalytic process; thus, revealing dual functionality of the protonated TA-COFs as both photodegradation and Brønsted acid catalysts. This pioneering work opens new avenues for harnessing the potential of the tetrazine linker in COF-based materials, facilitating advances in catalysis, sensing, and other related fields.

Reversible disassembly-reassembly of C-phycocyanin in pressurization-depressurization cycles of high hydrostatic pressure.

Hydrostatic pressure can reversibly modulate protein-protein and protein-chromophore interactions of C-phycocyanin (C-PC) from Spirulina platensis. Small-angle X-ray scattering combined with UV-Vis spectrophotometry and protein modeling was used to explore the color and structural changes of C-PC under high pressure conditions at different pH levels. It was revealed that pressures up to 350 MPa were enough to fully disassemble C-PC from trimers to monomers at pH 7.0, or from monomers to detached subunits at pH 9.0. These disassemblies were accompanied by protein unfolding that caused these high-pressure induced structures to be more extended. These changes were reversible following depressurization. The trimer-to-monomer transition proceeded through a collection of previously unrecognized, L-shaped intermediates resembling C-PC dimers. Additionally, pressurized C-PC showed decayed Q-band absorption and fortified Soret-band absorption. This was evidence that the folded tetrapyrroles, which had folded at ambient pressure, formed semicyclic unfolded conformations at a high pressure. Upon depressurization, the peak intensity and shift all recovered stepwise, showing pressure can precisely manipulate C-PC's structure as well as its color. Overall, a protein-chromophore regulatory theory of C-PC was unveiled. The pressure-tunability could be harnessed to modify and stabilize C-PC's structure and photochemical properties for designing new delivery and optical materials.

Mechanism and kinetics of enzymatic degradation of polyester microparticles using a shrinking particle-shrinking core model.

Generalized shrinking particle (SPM) and shrinking core (SCM) models were developed to the kinetics of heterogenous enzymatic degradation of polymer microparticles in a continuous microflow system. This enzymatic degradation was performed in a microfluidic device designed to both physically separate and immobilize the microparticles. Then time-resolved measurements were made using image processing of the physical changes of the particles during degradation. The kinetics of enzyme-polymer intermediate formation, enzymatic bond cleavage, and enzyme diffusion through the layer of degraded substrate (SCM only) were mathematically derived to predict the time-resolved degradation of the substrate. The proposed models were tested against the degradation of 15-25 µm particles of polycaprolactone (PCL) and poly (butylene adipate-co-terephthalate) (PBAT) by cutinase enzyme from Humicola insolens. Degradation of PCL microparticles followed the SPM model and its kinetics were found to be zero-order, while the SCM model applied to PBAT microparticles showed first-order kinetics. Further, the degradation of polybutylene succinate (PBS), and poly butylene-sebacate-co-terephthalate (PBSeT) microparticles demonstrated wide applicability of the method. The use of image processing simplifies the required analysis by eliminating the need to remove aliquots or concentrate effluent for additional analytical characterization.

Synthesis of a Cation-Exchange Resin by Inverse Suspension Polymerization for Lactoferrin Extraction from Whey.

Lactoferrin (LF), the main iron-binding protein of milk, has important nutritional, biological, and pharmaceutical properties. It is an essential nutritional component of newborn diets and also for adult health. Small amounts of lactoferrin can be found in whey, a nutritionally and biologically useful byproduct of the dairy industry. Although the amount of lactoferrin in whey is less than that in other sources like milk and bovine colostrum, the extraction of LF from this underused source has many economic and environmental benefits. The most common technique for the isolation of LF from dairy products is the use of cation-exchange resins. Here, we present the synthesis of a strong cation-exchange resin for the extraction of high-purity lactoferrin from whey. This resin was synthesized by inverse suspension copolymerization of aqueous solutions of sodium 2-acrylamido-2-methyl-1-propanesulfonate and N,N-methylenebisacrylamide in corn oil. The adsorption efficiency of this resin showed selective extraction of lactoferrin from four different whey sources. The adsorption efficiency of lactoferrin from these whey samples ranged from 93.8 to 97.4%.

Encapsulation and stabilization of lactoferrin in polyelectrolyte ternary complexes.

Effective delivery of the bioactive protein, lactoferrin (LF), remains a challenge as it is sensitive to environmental changes and easily denatured during heating, restricting its application in functional food products. To overcome these challenges, we formulated novel polyelectrolyte ternary complexes of LF with gelatin (G) and negatively charged polysaccharides, to improve the thermal stability of LF with retained antibacterial activity. Linear, highly charged polysaccharides were able to form interpolymeric complexes with LF and G, while coacervates were formed with branched polysaccharides. A unique multiphase coacervate was observed in the gum Arabic GA-LF-G complex, where a special coacervate-in-coacervate structure was found. The ternary complexes made with GA, soy soluble polysaccharide (SSP), or high methoxyl pectin (HMP) preserved the protein structures and demonstrated enhanced thermal stability of LF. The GA-LF-G complex was especially stable with >90% retention of the native LF after treatment at 90 °C for 2 min in a water bath or at 145 °C for 30 s, while the LF control had only ~ 7% undenatured LF under both conditions. In comparison to untreated LF, LF in ternary complex retained significant antibacterial activity on both Gram-positive and Gram-negative bacteria, even after heat treatment. These ternary complexes of LF maintain the desired functionality of LF, thermal stability and antibacterial activity, in the final products. The ternary complex structure, particularly the multiphase coacervate, may serve as a template for the encapsulation and stabilization of other bioactives and peptides.

Thermal Degradation of Thaumatin at Low pH and Its Prevention Using Alkyl Gallates.

Thaumatin, a potent sweet tasting protein extracted from the Katemfe Plant, is emerging as a natural alternative to synthetic non-nutritive sweeteners and flavor enhancer. As a food additive, its stability within the food matrix during thermal processing is of great interest to the food industry. When heated under neutral or basic conditions, thaumatin was found to lose its sweetness due to protein aggregation caused by sulfhydryl catalyzed disulfide bond interchange. At lower pH, while thaumatin was also found to lose sweetness after heating, it does so at a slower rate and shows more resistance to sweetness loss. SDS-PAGE indicated that thaumatin fragmented into multiple smaller pieces under heating in acidic pH. Using BEMPO-3, a lipophilic spin trap, we were able to detect the presence of a free-radical within the hydrophobic region of the protein during heating. Protein carbonyl content, a byproduct of protein oxidation, also increased upon heating, providing additional evidence for protein cleavage by a radical pathway. Hexyl gallate successfully inhibited the radical generation as well as protein carbonyl formation of thaumatin during heating.

Understanding animal-based flavor generation, mechanisms and characterization: Cheddar cheese and bacon flavors.

Natural animal-based flavors have great appeal to consumers and have broad applications in the food industry. In this review, we summarized findings related to bacon and Cheddar cheese flavors' components and their precursors, reaction mechanisms, influential factors, and characterization methods. The results show that free sugars, free amino acids, peptides, vitamins, lipids, and nitrites are precursors to bacon flavor. The conditions governing the formation of bacon flavor are thermally dependent, which facilitates the use of thermal food processing to generate such a flavor. For Cheddar cheese flavor, milk ingredients such as lactose, citrate, fat, and casein are reported as precursors. The optimum conditions to generate Cheddar cheese flavor from precursors are quite strict, which limits its application in food processing. As an alternative, it is more practical to generate Cheddar cheese flavor by combining key aroma compounds using thermal food processing. This review provides the food industry the comprehensive information about the generation of bacon and Cheddar cheese flavors using precursor molecules.

Biphasic Chemokinesis of Mammalian Sperm.

The female reproductive tract (FRT) continuously modulates mammalian sperm motion by releasing various clues as sperm migrate toward the fertilization site. An existing gap in our understanding of sperm migration within the FRT is a quantitative picture of how sperm respond to and navigate the biochemical clues within the FRT. In this experimental study, we have found that in response to biochemical clues, mammalian sperm display two distinct chemokinetic behaviors which are dependent upon the rheological properties of the media: chiral, characterized by swimming in circles; and hyperactive, characterized by random reorientation events. We used minimal theoretical modeling, along with statistical characterization of the chiral and hyperactive trajectories, to show that the effective diffusivity of these motion phases decreases with increasing concentration of chemical stimulant. In the context of navigation this concentration dependent chemokinesis suggests that the chiral or hyperactive motion refines the sperm search area within different FRT functional regions. Further, the ability to switch between phases indicates that sperm may use various stochastic navigational strategies, such as run and tumble or intermittent search, within the fluctuating and spatially heterogeneous environment of the FRT.

Lithium Pyrene Squarate Covalent Organic Frameworks for Efficient Lithium and Magnesium Separation from Salt Water.

The increasing pressure for lithium resources from the electric vehicle and nuclear energy industries means that new technologies to separate Mg2+ from Li+ from salt water are in demand. To address this need, we fabricated lithium pyrene squarate covalent organic frameworks (Li-SQCOFs) to separate Mg2+/Li+ mixtures from salt water. We optimized the effect of the electrolyte and the amount of the adsorbent and then carried out a kinetics study on the adsorbent recovery at various pH levels using both batch and continuous flow adsorption methods. Li-SQCOF was found to have excellent selectivity for solutions containing a mixture of Mg2+/Li+ ions. This work represents a unique path for the separation of Mg2+/Li+ through direct adsorption using a covalent organic framework (COF). The COF-supported ultrafiltration bed made in this study gave a Mg2+ separation flux of 60.5 h-1 m-2.

3D printing of responsive chiral photonic nanostructures.

Finely controlled flow forces in extrusion-based additive manufacturing can be exploited to program the self-assembly of malleable nanostructures in soft materials by integrating bottom-up design into a top-down processing approach. Here, we leverage the processing parameters offered by direct ink-writing (DIW) to reconfigure the photonic chiral nematic liquid crystalline phase in hydroxypropyl cellulose (HPC) solutions prior to deposition on the writing substrate to direct structural evolution from a particular initial condition. Moreover, we incorporate polyethylene glycol (PEG) into iridescent HPC inks to form a physically cross-linked network capable of inducing kinetic arrest of the cholesteric/chiral pitch at length scales that selectively reflect light throughout the visible spectrum. Based on thorough rheological measurements, we have found that printing the chiral inks at a shear rate where HPC molecules adopt pseudonematic state results in uniform chiral recovery following flow cessation and enhanced optical properties in the solid state. Printing chiral inks at high shear rates, on the other hand, shifts the monochromatic appearance of the extruded filaments to a highly angle-dependent state, suggesting a preferred orientation of the chiral domains. The optical response of these filaments when exposed to mechanical deformation can be used in the development of optical sensors.

One-Step Oxidation of Orange Peel Waste to Carbon Feedstock for Bacterial Production of Polyhydroxybutyrate.

Orange peels are an abundant food waste stream that can be converted into useful products, such as polyhydroxyalkanoates (PHAs). Limonene, however, is a key barrier to building a successful biopolymer synthesis from orange peels as it inhibits microbial growth. We designed a one-pot oxidation system that releases the sugars from orange peels while eliminating limonene through superoxide (O2• -) generated from potassium superoxide (KO2). The optimum conditions were found to be treatment with 0.05 M KO2 for 1 h, where 55% of the sugars present in orange peels were released and recovered. The orange peel sugars were then used, directly, as a carbon source for polyhydroxybutyrate (PHB) production by engineered Escherichia coli. Cell growth was improved in the presence of the orange peel liquor with 3 w/v% exhibiting 90-100% cell viability. The bacterial production of PHB using orange peel liquor led to 1.7-3.0 g/L cell dry weight and 136-393 mg (8-13 w/w%) ultra-high molecular weight PHB content (Mw of ~1900 kDa) during a 24 to 96 h fermentation period. The comprehensive thermal characterization of the isolated PHBs revealed polymeric properties similar to PHBs resulting from pure glucose or fructose. Our one-pot oxidation process for liberating sugars and eliminating inhibitory compounds is an efficient and easy method to release sugars from orange peels and eliminate limonene, or residual limonene post limonene extraction, and shows great promise for extracting sugars from other complex biomass materials.

Screening the Degradation of Polymer Microparticles on a Chip.

Enzymatic degradation of polymers has advantages over standard degradation methods, such as soil burial and weathering, which are time-consuming and cannot provide time-resolved observations. We have developed a microfluidic device to study the degradation of single microparticles. The enzymatic degradation of poly (1,4-butylene adipate-co-terephthalate) (PBAT) microparticles was studied using Novozym 51032 cutinase. PBAT microparticles were prepared via an oil-in-water emulsion solvent removal method, and their morphology and chemical composition were characterized. Then, microparticles with varying diameters of 30-60 µm were loaded into the microfluidic chip. Enzyme solutions at different concentrations were introduced to the device, and changes in the size and transparency of PBAT microparticles were observed over time. The physicochemical properties of degraded products were analyzed by FT-IR, NMR, mass spectrometry, and differential scanning calorimetry. The degradation process was also performed in bulk, and the results were compared to those of the microfluidic method. Our analysis confirms that the degradation process in both bulk and microfluidic methods was similar. In both cases, degradation takes place on aliphatic and soft segments of PBAT. Our findings serve as a proof of concept for a microfluidic method for easy and time-resolved degradation analysis, with degradation results comparable to those of conventional bulk methods.

Composition and functional properties of hemp seed protein isolates from various hemp cultivars.

Hemp seed protein isolates (HPI) were extracted from seven commercial hemp cultivars, a Cornell breeding line, and a commercial hemp heart product, and their composition and functional properties were investigated. HPI contained different ratios of edestin, vicilin, and albumin proteins, which affected protein solubility and functionality. Higher protein solubility was associated with cultivars that contained more vicilin and albumin, which influenced the subsequent functional properties of HPI. Significant differences in water holding capacity (0.83-1.05 g water/g protein isolate), oil holding capacity (1.28-1.81 g oil/g protein isolate), foam capacity (52.9%-84.9%), and foam stability (68.1%-89.4%) were observed across HPI. The Cornell hemp breeding line exhibited the highest protein solubility at pH 7.0 and was uniquely capable of forming an emulsion. The relationship identified between hemp seed protein composition and functionality, in conjunction with the demonstration of an on-going hemp breeding line, suggest that continued, targeted development of hemp cultivars can improve its seed protein functional properties for ingredient utilization in plant-based foods.

Starch Janus Particles: Bulk Synthesis, Self-Assembly, Rheology, and Potential Food Applications.

Although incredible progress in the field of Janus particles over the last three decades has delivered many promising smart-material prototypes, from cancer-targeting drug delivery vehicles to self-motile nanobots, their real-world applications have been somewhat tempered by concerns over scalability and sustainability. In this study, we adapt a simple, scalable 3D mask method to synthesize Janus particles in bulk using starch as the base material: a natural biopolymer that is safe, biocompatible, biodegradable, cheap, widely available, and versatile. Using this method, starch granules are first embedded on a wax droplet such that half of the starch is covered; then, the uncovered half is treated with octenyl succinic anhydride, after which the wax coating is removed. Janus particles with 49% Janus balance can be produced in this way and were observed to self-assemble into wormlike strings in water due to their hydrophobic/hydrophilic nature. Our Janus starch granules outperform the non-Janus controls as thickening and gelling agents: they exhibit a fourfold increase in water-holding capacity, a 30% lower critical caking concentration, and a viscosity greater by orders of magnitude. They also form gels that are much firmer and more stable. Starch Janus particles with these functional properties can be used as novel, lower-calorie, highly efficient, plant-based super-thickeners in the food industry, potentially reducing starch use in food by 55%.