Structured soft particulate matters for delivery of bioactive compounds in foods and functioning in the colon.

The present review discusses challenges, perspectives, and current needs of delivering bioactive compounds (BCs) using soft particulate matters (SPMs) for gut health. SPMs can entrap BCs for incorporation in foods, preserve their bioactivities during processing, storage, and gastrointestinal digestion, and deliver BCs to functioning sites in the colon. To enable these functions, physical, chemical, and biological properties of BCs are integrated in designing various types of SPMs to overcome environmental factors reducing the bioavailability and bioactivity of BCs. The design principles are applied using food grade molecules with the desired properties to produce SPMs by additionally considering the cost, sustainability, and scalability of manufacturing processes. Lastly, to make delivery systems practical, impacts of SPMs on food quality are to be evaluated case by case, and health benefits of functional foods incorporated with delivery systems are to be confirmed and must outweigh the cost of preparing SPMs.

Drying of probiotics to enhance the viability during preparation, storage, food application, and digestion: A review.

Functional food products containing viable probiotics have become increasingly popular and demand for probiotic ingredients that maintain viability and stability during processing, storage, and gastrointestinal digestions. This has resulted in heightened research and development of powdered probiotic ingredients. The aim of this review is to overview the development of dried probiotics from upstream identification to downstream applications in food. Free probiotic bacteria are susceptible to various environmental stresses during food processing, storage, and after ingestion, necessitating additional materials and processes to preserve their activity for delivery to the colon. Various classic and emerging thermal and nonthermal drying technologies are discussed for their efficiency in preparing dehydrated probiotics, and strategies for enhancing probiotic survival after dehydration are highlighted. Both the formulation and drying technology can influence the microbiological and physical properties of powdered probiotics that are to be characterized comprehensively with various techniques. Furthermore, quality control during probiotic manufacturing and strategies of incorporating powdered probiotics into liquid and solid food products are discussed. As emerging technologies, structure-design principles to encapsulate probiotics in engineered structures and protective materials with improved survivability are highlighted. Overall, this review provides insights into formulations and drying technologies required to supplement viable and stable probiotics into functional foods, ensuring the retention of their health benefits upon consumption.

Selection of mutant Listeria phages under food-relevant conditions can enhance application potential.

Bacteriophages are viruses that infect and kill bacteria. Currently, phage products are available for the control of the pathogen Listeria monocytogenes in food products in the United States. In this study, we explore whether experimental evolution can be used to generate phages with improved abilities to function under specific food-relevant conditions. Ultra-pasteurized oat and whole milk were chosen as test matrices as they represent different food groups, yet have similar physical traits and macronutrient composition. We showed that (i) wild-type phage LP-125 infection kinetics are different in the two matrices and (ii) LP-125 has a significantly higher burst size in oat milk. From this, we attempted to evolve LP-125 to have improved infection kinetics in whole milk. Ancestral LP-125 was passaged through 10 rounds of amplification in milk conditions. Plaque-purified DNA samples from milk-selected phages were isolated and sequenced, and mutations present in the isolated phages were identified. We found two nonsynonymous substitutions in LP125_108 and LP125_112 genes, which encode putative baseplate-associated glycerophosphoryl diester phosphodiesterase and baseplate protein, respectively. Protein structural modeling showed that the substituted amino acids in the mutant phages are predicted to localize to surface-exposed helices on the corresponding structures, which might affect the surface charge of proteins and their interaction with the bacterial cell. The phage containing the LP125_112 mutation adsorbed significantly faster than the ancestral phage in both oat and whole milk. Follow-up experiments suggest that fat content may be a key factor for the expression of the phenotype of this mutation. IMPORTANCE Bacteriophages are one of the tools available to control the foodborne pathogen, Listeria monocytogenes. Phage products must work under a broad range of food conditions to be an effective control for L. monocytogenes. Here, we show that the experimental evolution of phages can be used to generate new phages with phenotypes useful under specific conditions. We used this approach to select for a mutant phage that more efficiently binds to L. monocytogenes that is grown in whole milk and oat milk. We show that the fat content of these milks is necessary for the expression of this phenotype. Our findings show that experimental evolution can be used to select for improved phages with better performance under specific conditions. This approach has the potential to support the development of condition-specific phage-based biocontrols in the food industry.

A drug-free nanozyme for mitigating oxidative stress and inflammatory bowel disease.

Inflammatory bowel disease (IBD) is an incurable disease of the gastrointestinal tract with a lack of effective therapeutic strategies. The proinflammatory microenvironment plays a significant role in both amplifying and sustaining inflammation during IBD progression. Herein, biocompatible drug-free ceria nanoparticles (CeNP-PEG) with regenerable scavenging activities against multiple reactive oxygen species (ROS) were developed. CeNP-PEG exerted therapeutic effect in dextran sulfate sodium (DSS)-induced colitis murine model, evidenced by corrected the disease activity index, restrained colon length shortening, improved intestinal permeability and restored the colonic epithelium disruption. CeNP-PEG ameliorated the proinflammatory microenvironment by persistently scavenging ROS, down-regulating the levels of multiple proinflammatory cytokines, restraining the proinflammatory profile of macrophages and Th1/Th17 response. The underlying mechanism may involve restraining the co-activation of NF-κB and JAK2/STAT3 pathways. In summary, this work demonstrates an effective strategy for IBD treatment by ameliorating the self-perpetuating proinflammatory microenvironment, which offers a new avenue in the treatment of inflammation-related diseases.

Inactivation of Escherichia coli K12 on raw almonds using supercritical carbon dioxide and thyme oil.

Raw almonds could be contaminated by pathogens, but the current pasteurization practice using propylene oxide in the U.S. has flammability and carcinogenicity concerns. Supercritical carbon dioxide (scCO2) is a water-free technology and is a solvent of essential oils that are effective antimicrobial preservatives. The objective of this study was to investigate the possibility of combining scCO2 and thyme oil (TO) to reduce Escherichia coli K12 inoculated on raw almonds. Raw almonds inoculated with ∼6 log CFU/g E. coli K12 were batch-treated with scCO2 alone or the combination of presoaking in pure TO followed by scCO2 treatments at different combinations of temperature, pressure, and duration. Compared to scCO2 alone treatments, the combination of TO and scCO2 treatments significantly improved the disinfection effectiveness. Temperature had the most significant effect on the log reduction. At 70 °C, the log reduction by the combination treatment was over 4-log CFU/g and the maximum reduction was 5.16 log CFU/g. The findings suggest that the combination of TO and scCO2 may be a potential water-free technology to meet the requirement of over 4-log reduction of target microorganism for almond and other tree nut products.

Physical and sensory properties of lemon-flavored acidic beverages formulated with nonfat dry milk during storage.

Sensory and physical properties of 2 lemon-flavored beverages with 5% and 7.5% wt/wt nonfat dry milk (NFDM) at pH 2.5 were studied during storage. The 2 beverages had similar volatile compounds, but the 5% NFDM had higher aroma and lemon flavor, with a preferred appearance by consumers due to the lower turbidity and viscosity. After 28 d of storage at 4°C, lemon flavor decreased in the 5% NFDM beverage but was still more intense than the 7.5% one. During 70 d of storage, no microorganisms were detected, and the beverages were more stable when stored at 4°C than at room temperature according to changes of physical properties measured for appearance, turbidity, color, particle size, zeta potential, rheological properties, and transmission electron microscopy morphology. Findings of the present study suggest that NFDM may be used at 5% wt/wt to produce stable acidic dairy beverages with low turbidity when stored at 4°C.

Conjugation of α-, ß-, and κ-Caseins with Propylene Glycol Alginate Using a Transacylation Reaction as Novel Emulsifiers.

Protein-polysaccharide conjugates usually have better emulsifying properties than their constituent biopolymers. In this study, casein-alginate conjugates were prepared using a transacylation reaction between different types of caseins and propylene glycol alginate (PGA) at pH 11.0. The purified conjugates had the combined molecular structure of casein and hydrolyzed PGA (alginate), according to Fourier-transform infrared spectroscopy, and α-, ß-, and κ-casein-alginate conjugates had 39.17%, 37.78%, and 23.14% protein content, respectively. Emulsions were prepared with all the conjugates at a surfactant-to-oil ratio of 1:100 (w: (v), which is much lower than emulsions stabilized by Maillard-type protein-polysaccharide conjugates. The emulsions had high stability during storage and at wide pH (3.0-11.0) and ionic strength (0-450 mM) ranges. The interfacial tension of conjugate dispersions and therefore the droplet size were dependent on the protein content, not the casein type, while the polysaccharide moiety was critical to the emulsion stability. The present findings suggest a transacylation reaction can be used to prepare protein-alginate conjugates with novel emulsifying properties.

Physicochemical properties of skim milk powder dispersions prepared with calcium-chelating sodium tripolyphosphate, trisodium citrate, and sodium hexametaphosphate.

Due to health benefits of proteins, the demand for protein beverages has grown rapidly. Translucent protein drinks with neutral pH may have advantages over acidic beverages that may cause dental erosion, and skim milk powder (SMP) is an affordable protein ingredient. Dissociating casein micelles by calcium chelators is a well-known method to reduce SMP dispersion turbidity, but much is to be studied for physicochemical properties as affected by chelator type and concentration. The objective of the present study was to characterize physicochemical properties of dispersions with 5% (wt/vol) SMP after addition of 0 to 30 mM sodium tripolyphosphate, trisodium citrate, or sodium hexametaphosphate. The turbidity was decreased with increasing chelator concentration, with the lowest turbidity observed in the SMP dispersions with sodium hexametaphosphate. The smallest hydrodynamic diameter was observed at an intermediate chelator concentration, resulting from the balance of casein micelle dissociation and aggregation of dissociated caseins induced at an elevated ionic strength. Heating at 90°C for 5 min increased turbidity but lowered hydrodynamic diameter of SMP dispersions, with some exceptions. The morphology of SMP dispersions differed for each chelator and was also affected by chelator concentration and heating. Trisodium citrate was the most effective to demineralize colloidal calcium phosphate in casein micelles, but the amount of dissolved calcium was not directly correlated with the decreased turbidity, indicating different chelating mechanisms by each chelator. Analysis of serum calcium and phosphorus concentrations also suggested that the type and concentration of soluble and insoluble calcium phosphates and their partitioning in the serum and casein micelles were dynamically changed by the studied parameters to affect dispersion turbidity and structures of casein micelles. Findings from the present study may be used to formulate translucent beverages incorporating SMP and other casein micelle ingredients.

Inhibiting Ice Recrystallization by Nanocelluloses.

Biocompatible materials with ice recrystallization inhibition (IRI) activity have potential applications in several fields. Emerging studies have associated the IRI activity of antifreeze proteins/glycoproteins and several mimics of synthetic materials with a facially amphipathic structure. Nanocelluloses are a new family of renewable materials that demonstrate amphiphilicity. Herein the IRI activity of cellulose nanocrystals (CNCs) and 2,2,6,6-tetramethylpiperidine-1-oxyl oxidized cellulose nanofibrils (TEMPO-CNFs) is reported. In 0.01 M NaCl, ice recrystallization was effectively inhibited by 5.0 mg/mL CNCs or 2.0 mg/mL TEMPO-CNFs. In phosphate-buffered saline, observable IRI activity was found with 30.0 mg/mL CNCs. IRI assays in sucrose solutions showed that the decreased IRI activity of nanocelluloses in saline was caused by the aggregation of nanocelluloses due to charge screening. Neither thermal hysteresis nor dynamic ice shaping activity was observed in nanocelluloses. These findings may lead to the use of nanocelluloses as novel ice recrystallization inhibitors.

Potential of acidified sodium benzoate as an alternative wash solution of cherry tomatoes: Changes of quality, background microbes, and inoculated pathogens during storage at 4 and 21°C post-washing.

To evaluate the feasibility of acidified sodium benzoate (NaB) as alternative washing solutions of fresh produce, the survival of inoculated pathogens, the background molds and yeasts counts, and quality parameters were compared during 4 and 21 °C storage of cherry tomatoes washed with 3000 ppm NaB at pH 2.0, 200 ppm free chlorine at pH 6.5, water adjusted to pH 2.0, and distilled water. The acidified NaB solution was the most effective in reducing the population of Escherichia coli O157:H7, Salmonella enterica and Listeria monocytogenes cocktails on tomatoes (>4 log CFU/g). NaB was more effective than free chlorine (P < 0.05) in reducing the two Gram-negative bacteria on tomatoes, while the reduction of Gram-positive L. monocytogenes by NaB (5.49 log CFU/g) and chlorine (4.98 log CFU/g) was similar (P > 0.05). No recovery of bacteria was found in all treatments during storage for 15 days. The acidified NaB effectively controlled yeasts and molds on cherry tomatoes to <1 log CFU/g or below the detection limit at both temperatures during 15-d storage, while free chlorine did not. Compared to unwashed controls, NaB had no effect on color, weight loss, firmness, and total soluble solids content of tomatoes during storage. The effect of NaB reducing pathogenic and spoilage microorganisms on tomatoes and maintaining quality during storage suggests its potential as an alternative wash solution in postharvest processing of fresh produce.

Properties of Ternary Biopolymer Nanocomplexes of Zein, Sodium Caseinate, and Propylene Glycol Alginate and Their Functions of Stabilizing High Internal Phase Pickering Emulsions.

A pH-cycle method based on preparing an alkaline solution of zein followed by neutralization with an acid can be used to prepare zein nanoparticles. In the present work, partial alkaline hydrolysis of propylene glycol alginate (PGA) to a lower pH was studied to prepare binary zein-PGA nanocomplexes and ternary complexes with additional sodium caseinate (NaCas). 0.5% or more PGA was sufficient to reduce the pH to 7.5 or lower, eliminating the need for titration, and resulted in simultaneous nanocomplex formation. The addition of NaCas into alkaline zein-PGA solution resulted in smaller complexes with all biopolymers, whereas adsorption on binary zein-PGA complexes was observed when NaCas was added into the neutral zein-PGA dispersions. The formation of nanocomplexes involved with hydrophobic and electrostatic attractions and hydrogen bonds and was further affected by the amount of NaCas. The ternary nanocomplexes with equal masses of zein and NaCas had an excellent capacity to prepare gel-like Pickering emulsions with as much as 80% v/v oil with characteristics suitable for texture modification and delivery systems of bioactive compounds in food and consumer products. Therefore, PGA can be used to possibly scale-up the pH-cycle method to produce zein-based nanoparticles with unique functional properties.

Core-Shell Nanoencapsulation of α-Tocopherol by Blending Sodium Oleate and Rebaudioside A: Preparation, Characterization, and Antioxidant Activity.

Nanoencapsulation of α-tocopherol (α-TOC) by blending sodium oleate (NaOl) and rebaudioside A (RebA) was successfully prepared by self-assembly method under mild conditions. The optimized nanoemulsion showed the loading capacity of α-TOC was 30 wt% of sodium oleate. FTIR analysis suggested that hydrogen bonds and hydrophobic interactions were the major forces in α-TOC-NaOl/RebA complexes that were spherical and possessed well-distinguishable core-shell structures. The freeze-dried α-TOC-NaOl/RebA complexes had great stability under ambient conditions. The release profile of α-TOC showed a first-order kinetics reaching around 67.9% after 90 h at 25 °C. Nanoencapsulation improved dispersibility and greatly increased the antioxidant activity of α-TOC. Therefore, the stable α-TOC-NaOl/RebA core-shell complexes prepared from "generally recognized as safe" (GRAS) ingredients have great potential to supplement α-TOC in food and cosmetic products.

Evaporation-induced self-assembly of quantum dots-based concentric rings on polymer-based nanocomposite films.

The "ball-on-film" template is used to construct concentric rings on the surface of PMMA-QDs (polymethyl methacrylate - quantum dots) nanocomposite films via the evaporation of pure chloroform droplets, which are confined by a steel ball. The concentric rings consist of QDs, as revealed by the fluorescence images of the concentric rings. The photoluminescence intensity of the concentric rings increases with the increase of the distance to the ball center, suggesting that the amount of QDs accumulated around the contact line at individual stick state increases with the increase of the distance to the ball center. Both the wavelength and cross-sectional area (width) of the concentric rings increase approximately linearly with increasing distance to the ball center, independent of the ball size, the film thickness and the QDs concentration. For the PMMA-QDs nanocomposite films prepared from the same QDs concentration in chloroform, the thicker the PMMA-QDs nanocomposite film, the larger the wavelength for the same distance to the ball center. The effect of confinement of two steel balls on the surface patterns over the PMMA-QDs nanocomposite films is studied via a template of "two spheres on film". Symmetric surface patterns are formed. There exist two types of featureless zone between the two balls, depending on the distance between the two balls: one is the inner featureless zone and the other is the outer featureless zone. The size of both featureless zones increases with the increase of the ball distance.

Amyloid-like fibrils formed from intrinsically disordered caseins: physicochemical and nanomechanical properties.

Amyloid-like fibrils are studied because of their significance in understanding pathogenesis and creating functional materials. Amyloid-like fibrils have been studied by heating globular proteins at acidic conditions. In the present study, intrinsically disordered α-, ß-, and κ-caseins were studied to form amyloid-like fibrils at pH 2.0 and 90 °C. No fibrils were observed for α-caseins, and acid hydrolysis was found to be the rate-limiting step of fibrillation of ß- and κ-caseins. An increase of ß-sheet structure was observed after fibrillation. Nanomechanic analysis of long amyloid-like fibrils using peak-force quantitative nanomechanical atomic force microscopy showed the lowest and highest Young's modulus for ß-casein (2.35 ± 0.29 GPa) and κ-casein (4.14 ± 0.66 GPa), respectively. The dispersion with ß-casein fibrils had a viscosity more than 10 and 5 times higher than those of κ-casein and ß-lactoglobulin, respectively, at 0.1 s(-1) at comparable concentrations. The current findings may assist not only the understanding of amyloid fibril formation but also the development of novel functional materials from disordered proteins.

Decolorization of Cheddar cheese whey by activated carbon.

Colored Cheddar whey is a source for whey protein recovery and is decolorized conventionally by bleaching, which affects whey protein quality. Two activated carbons were studied in the present work as physical means of removing annatto (norbixin) in Cheddar cheese whey. The color and residual norbixin content of Cheddar whey were reduced by a higher level of activated carbon at a higher temperature between 25 and 55°C and a longer time. Activated carbon applied at 40g/L for 2h at 30°C was more effective than bleaching by 500mg/L of hydrogen peroxide at 68°C. The lowered temperature in activated-carbon treatments had less effect on protein structure as investigated for fluorescence spectroscopy and volatile compounds, particularly oxidation products, based on gas chromatography-mass spectrometry. Activated carbon was also reusable, removing more than 50% norbixin even after 10 times of regeneration, which showed great potential for decolorizing cheese whey.

Impacts of sample preparation methods on solubility and antilisterial characteristics of essential oil components in milk.

Essential oil components (EOCs) have limited water solubility and are used at much higher concentrations in complex food matrices than in growth media to inhibit pathogens. However, the correlation between solubility and activity has not been studied. The objective of this work was to characterize the solubility of EOCs in solvents and milk and correlate solubility with antilisterial activity. The solubilities of four EOCs, thymol, carvacrol, eugenol, and trans-cinnamaldehyde, in water was significantly increased in the presence of 5% (vol/vol) ethanol. In milk, the solubility of EOCs was lower than in water, with lower solubility in higher-fat milk. EOCs applied to milk as stock solutions (in 95% aqueous ethanol) enabled quicker dissolution and higher solubility in milk serum than other methods of mixing, such as end to end, and greater reductions of Listeria monocytogenes Scott A after 0 and 24 h. When the EOC concentration detected in milk serum was above the minimum bactericidal concentration, complete inhibition of L. monocytogenes in tryptic soy broth resulted. Therefore, the antilisterial properties in milk could be correlated with the solubility by comparison to the minimum inhibitory or bactericidal concentrations of EOCs. While the EOCs applied using ethanol generally had solubility and activity characteristics superior to those of other mixing methods, ethanol is not used to a great extent in nonfermented foods. Therefore, mixing methods without an organic solvent may be more readily adaptable to enhancing the distribution of EOCs in complex food systems.

pH-driven encapsulation of curcumin in self-assembled casein nanoparticles for enhanced dispersibility and bioactivity.

The poor water solubility and bioactivity of lipophilic phytochemicals can be potentially improved by delivery systems. In this study, a low-cost, low-energy, and organic solvent-free encapsulation technology was studied by utilizing the pH-dependent solubility properties of curcumin and self-assembly properties of sodium caseinate (NaCas). Curcumin was deprotonated and dissolved, while NaCas was dissociated at pH 12 and 21 °C for 30 min. The subsequent neutralization enabled the encapsulation of curcumin in self-assembled casein nanoparticles. The degradation of curcumin under encapsulation conditions was negligible based on visible light and nuclear magnetic resonance spectroscopy. The dissociation of NaCas at pH 12 and reassociation after neutralization were confirmed using dynamic light scattering and analytical ultracentrifugation. The curcumin encapsulated in casein nanoparticles showed significantly improved anti-proliferation activity against human colorectal and pancreatic cancer cells. The studied encapsulation method is promising to utilize lipophilic compounds in food or pharmaceutical industries.

Removal of milk fat globules from whey protein concentrate 34% to prepare clear and heat-stable protein dispersions.

Whey protein concentrates (WPC) are low-cost protein ingredients, but their application in transparent ready-to-drink beverages is limited due to turbidity caused by fat globules and heat instability. In this work, fat globules were removed from WPC 34% (WPC-34) to prepare heat-stable ingredients via the Maillard reaction. The removal of fat globules by acid precipitation and centrifugation was observed to be the most complete at pH 4.0, and the loss of protein was caused by micrometer-sized fat globules and protein aggregates. Spray-dried powder prepared from the transparent supernatant was glycated at 130°C for 20 and 30min or 60°C for 24 and 48h. The 2 groups of samples had comparable heat stability and degree of glycation, evaluated by free amino content and analytical ultracentrifugation, but high-temperature, short-time treatment reduced the color formation during glycation. Therefore, WPC-34 can be processed for application in transparent beverages.

Engineered Nanoscale Food Ingredients: Evaluation of Current Knowledge on Material Characteristics Relevant to Uptake from the Gastrointestinal Tract.

The NanoRelease Food Additive project developed a catalog to identify potential engineered nanomaterials (ENMs) used as ingredients, using various food-related databases. To avoid ongoing debate on defining the term nanomaterial, NanoRelease did not use any specific definition other than the ingredient is not naturally part of the food chain, and its dimensions are measured in the nanoscale. Potential nanomaterials were categorized based on physical similarity; analysis indicated that the range of ENMs declared as being in the food chain was limited. Much of the catalog's information was obtained from product labeling, likely resulting in both underreporting (inconsistent or absent requirements for labeling) and/or overreporting (inability to validate entries, or the term nano was used, although no ENM material was present). Three categories of ingredients were identified: emulsions, dispersions, and their water-soluble powdered preparations (including lipid-based structures); solid encapsulates (solid structures containing an active material); and metallic or other inorganic particles. Although much is known regarding the physical/chemical properties for these ingredient categories, it is critical to understand whether these properties undergo changes following their interaction with food matrices during preparation and storage. It is also important to determine whether free ENMs are likely to be present within the gastrointestinal tract and whether uptake of ENMs may occur in their nanoform physical state. A practical decision-making scheme was developed to help manage testing requirements.

Enhancing the functionality of pea proteins by conjugation with propylene glycol alginate via transacylation reaction assisted with ultrasonication.

Pea proteins lack the desirable functional characteristics for food and beverage applications. In this study, transacylation reaction assisted with ultrasonication was used to glycate pea proteins with propylene glycol alginate to enhance their functional properties. The reaction was carried out at pH 11.0 for different pea protein isolate: propylene glycol alginate mass ratios and time durations in a sonic bath at 40 °C. Glycation was confirmed in gel electrophoresis, and ultrasonication enhanced the glycation, with optimal degrees of glycation observed at 45 min reaction time and mass ratios of 2:1 (37.73%) and 1:1 (35.96%). The transacylation reaction increased random coil content of pea proteins by 28% and enhanced their solubility by 2.02 times at pH 7.0, water holding capacity by >50% at pH 7.0, foaming properties, emulsifying properties, and heat stability. This study offers a novel approach that can enhance functionality and applicability of pea proteins.