Nonenzymatic glycation as a tunable technique to modify plant proteins: A comprehensive review on reaction process, mechanism, conjugate structure, and functionality.

Plant proteins are expected to become a major protein source to replace currently used animal-derived proteins in the coming years. However, there are always challenges when using these proteins due to their low water solubility induced by the high molecular weight storage proteins. One approach to address this challenge is to modify proteins through Maillard glycation, which involves the reaction between proteins and carbohydrates. In this review, we discuss various chemical methods currently available for determining the indicators of the Maillard reaction in the early stage, including the graft degree of glycation and the available lysine or sugar, which are involved in the very beginning of the reaction. We also provide a detailed description of the most popular methods for determining graft sites and assessing different plant protein structures and functionalities upon non-enzymatic glycation. This review offers valuable insights for researchers and food scientists in order to develop plant-based protein ingredients with improved functionality.

Traditional fermented soybean products: processing, flavor formation, nutritional and biological activities.

Traditional fermented soybean food has emerged as an important part of people's dietary structure because of the unique flavors and improved health benefit. During fermentation, the nutrients in soybean undergo a series of biochemical reactions catalyzed naturally by microorganism secreted enzymes. Thereafter, many functional and bioactive substances such as bioactive peptides, unsaturated fatty acids, free soy isoflavones, vitamins and minerals are produced, making fermented soy products more advantageous in nutrition and health. This review comprehensively discusses the historical evolution, distribution, traditional fermentation processing, main sources and characteristics of fermented strains, flavor components, nutritional properties, and biological activities of four traditional fermented soybean foods including douchi, sufu, dajiang, and soy sauce. In the end, we introduce four major challenges encountered by traditional fermented soybean foods including high salt content, formation of biogenic amine, the presence of pathogenic microorganisms and mycotoxins, and quality inconsistency. We conclude that the establishment of scientific quality standard and innovated fermentation processing is the potential solutions to combat the issues and improve the safety of traditional fermented soybean products.

Chemical Composition of Tomato Seed Flours, and Their Radical Scavenging, Anti-Inflammatory and Gut Microbiota Modulating Properties.

In the current study, the chemical composition and total phenolic content of tomato seed flours, along with potential health beneficial properties, including free radical scavenging capacities, anti-inflammatory capacities, and gut microbiota profile modulation, were examined using two different batches. Eight compounds were identified in the tomato seed flour, including malic acid, 2-hydroxyadipic acid, salicylic acid, naringin, N-acetyl-tryptophan, quercetin-di-O-hexoside, kaempferol-di-O-hexoside, and azelaic acid. The total phenolic contents of tomato seed flour were 1.97-2.00 mg gallic acid equivalents/g. Oxygen radical absorbing capacities (ORAC), 2,2-diphenyl-1-picrylhydrazyl radical scavenging capacities (DPPH), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) cation radical scavenging capacities (ABTS) were 86.32-88.57, 3.57-3.81, and 3.39-3.58 µmoles Trolox equivalents/g, respectively, on a per flour dry weight basis. The mRNA expression of the pro-inflammatory markers, interleukin-1 beta (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-α), were dose-dependently suppressed by tomato seed flour extracts. The extracts altered five of the eight bacterial phyla and genera evaluated. The results may provide some scientific support for the use of tomato seed flour as value-added food ingredients.

Modification of pulse proteins for improved functionality and flavor profile: A comprehensive review.

Consumers' preference to have a healthy eating pattern has led to an increasing demand for more nutrient-dense and healthier plant-based foods. Pulse proteins are exceptional quality ingredients with potential nutritional benefits, and might act as health-promoting agents for addressing the new-generation foods. However, the utilization of pulse protein in foods has been hampered by its relatively poor functionality and unpleasant flavor. Protein structure modification has been proved to be a useful means to improve the functionality and flavor profile of pulse protein. This paper begins with a brief introduction of hierarchical structure of pulse protein materials to better understand the structure characteristics. A comprehensive review is presented on the current techniques including chemical and enzymatic modifications and molecular breeding on pulse protein structure and functionality/flavor. The mechanism and the limitations and the toxicological concerns of these approaches are discussed. We conclude that understanding protein structure-functionality relationship is extremely valuable in tailoring proteins for specific functional outcomes and expanding the availability of pulse proteins. Furthermore, selective protein modification is a valuable in-depth toolkit for generating novel protein constructs with preferable functional attributes and flavor profiles. Innovative structure modification with special focus on the molecular basis for the exquisite protein designs is a pillar of pulse protein access to the desired functionality.

Phenolic compounds in germinated cereal and pulse seeds: Classification, transformation, and metabolic process.

Natural phenolic compounds are rich in cereal and pulse seeds and their dietary functions tend to improve dramatically during germination. This article reviews recent research on the transformation of phenolic compounds during seed germination. In particular, it highlights the classification of crude phenolic compounds that can be divided into extractable and non-extractable phenolic compounds based on the biosynthesis process and extraction method. It also recommends grouping resorcinol lipids in the category of extractable phenolic compounds as non-polar solvent extractable phenolic compounds. Moreover, it discusses the variation of the different form of phenolic compounds and proposes a possible metabolic model of these phenolic compounds for seeds germination. This article is crucial for phenolic compounds research, cereal and pulse seeds germination, and food ingredients industry.

Viability of Lactobacillus rhamnosus GG microencapsulated in alginate/chitosan hydrogel particles during storage and simulated gastrointestinal digestion: role of chitosan molecular weight.

Sodium alginate hydrogel particles coated with cationic biopolymers have been shown to be one of the promising means for probiotic encapsulation and protection. In this study, we aimed to systematically explore the effect of molecular weight of chitosan coating on the functional performance of sodium alginate hydrogel particles for improving the viability of Lactobacillus rhamnosus GG (LGG). We first electrostatically deposited three different molecular weights of chitosan coatings, i.e., chitosan oligosaccharide (COS), low molecular weight chitosan (LMW-chitosan) and medium molecular weight chitosan (MMW-chitosan) on sodium alginate hydrogel particles. Both SEM and FTIR results indicated that chitosan was successfully deposited onto the surface of the hydrogel particles. We then evaluated the effect of chitosan MW on the viability of LGG encapsulated in the hydrogels during long-term storage and simulated gastrointestinal digestion. Among them, the hydrogel particles coated with COS prevented the viability loss of LLG during long-term storage at different temperatures (4, 25 and 37 °C). However, we did not find any improvement in the viability of the encapsulated LGG by all three chitosan coatings during simulated digestion.

Occurrence and preventive strategies to control mycotoxins in cereal-based food.

Mycotoxins contamination in cereal-based food is ubiquitous according to systematic review of the scientific documentation of worldwide mycotoxin contamination in cereal and their products between 2008 and 2018, thus representing food safety issue especially in developing tropical countries. Food processing plays a vital role to prevent mycotoxin contamination in food. Therefore, it is with great urgency to develop strategies to inhibit fungi growth and mycotoxin production during food processing. This review begins by discussing physicochemical properties of five most common mycotoxins (aflatoxins, fumonisins, ochratoxins, deoxynivalenol, and zearalenone) found in cereal grains, regulation for mycotoxins in food, and their potential negative impact on human health. The fate of mycotoxins during major cereal-based food processing including milling, breadmaking, extrusion, malting, and brewing was then summarized. In the end, traditional mitigation strategies including physical and chemical and potential application of biocontrol agent and essential oil nanoemulsions that can be applied during food processing were discussed. It indicated that no single method is currently available to completely prevent mycotoxin contamination in cereal foods.

Ultrasensitive electrochemiluminescence immunosensor for the transcriptional co-activator p300 by using a graphene oxide monolayer and tetrahedral DNA-mediated signal amplification.

Protein p300 is a transcriptional co-activator that participates in many physiological processes including cell cycle control, differentiation and apoptosis. It serves (a) as a protein bridge that links specific transcription factors to the fundamental transcription machinery, (b) as a scaffold to complete multiple transcription cofactors, and (c) as an enzyme for acetylating histone and non-histone proteins. An ultrasensitive electrochemiluminescence (ECL) immunosensor is described here that is based on the use of a magnetic glassy carbon electrode modified with tetrahedral DNA with hollow structure, graphene oxide (GO) and gold nanocrystals. The use of a GO monolayer allows for greater carrying capacity and warrants a wider outer Helmholtz plane. Strong and stable ECL signals were achieved due to antigen-antibody interaction by using the ECL probe Ru(phen)32+. This immunosensor has a response that covers the 0.005 to 80 nM p300 concentration range and has a 1 pM detection limit. It was exploited for the determination of p300 in HeLa cell lysate and (spiked) serum. Graphical abstract Schematic presentation of an ultrasensitive Faraday-cage electrochemiluminescence immunosensor toward the transcriptional co-activator p300 analysis is presented based on a graphene oxide monolayer and tetrahedral DNA-mediated signal amplification.

Signal-on electrochemical assay for label-free detection of TdT and BamHI activity based on grown DNA nanowire-templated copper nanoclusters.

Electrochemical methods allow fast and inexpensive analysis of enzymatic activity. Here, a simple and yet efficient "signal-on" electrochemical assay for sensitive, label-free detection of DNA-related enzyme activity was established on the basis of terminal deoxynucleotidyl transferase (TdT)-mediated extension strategy. TdT, which is a template-independent DNA polymerase, can catalyze the sequential addition of deoxythymidine triphosphate (dTTP) at the 3'-OH terminus of single-stranded DNA (ssDNA); then, the TdT-yield T-rich DNA nanowires can be employed as the synthetic template of copper nanoclusters (CuNCs). Grown DNA nanowires-templated CuNCs (noted as DNA-CuNCs) were attached onto graphene oxide (GO) surface and exhibited unique electrocatalytic activity to H2O2 reduction. Under optimal conditions, the proposed biosensor was utilized for quantitatively monitoring TdT activity, with the observed LOD of 0.1 U/mL. It also displayed high selectivity to TdT with excellent stability, and offered a facile, convenient electrochemical method for TdT-relevant inhibitors screening. Moreover, the proposed sensor was successfully used for BamHI activity detection, in which a new 3'-OH terminal was exposed by the digestion of a phosphate group. Ultimately, it has good prospects in DNA-related enzyme-based biochemical studies, disease diagnosis, and drug discovery. Graphical Abstract Extraordinary TdT-generated DNA-CuNCs are synthesized and act as a novel electrochemical sensing platform for sensitive detection of TdT and BamHI activity in biological environments.

Plant protein solubility: A challenge or insurmountable obstacle.

Currently, there is an increasing focus on comprehending the solubility of plant-based proteins, driven by the rising demand for animal-free food formulations. The solubility of proteins plays a crucial role in impacting other functional properties of proteins and food processing. Consequently, understanding protein solubility in a deeper sense may allow a better usage of plant proteins. Herein, we discussed the definition of protein solubility from both thermodynamic and colloidal perspectives. A range of factors affecting solubility of plant proteins are generalized, including intrinsic factors (amino acids composition, hydrophobicity), and extrinsic factors (pH, ionic strength, extraction and drying methods). Current methods to enhance solubility are outlined, including microwave, high intensity ultrasound, hydrostatic pressure, glycation, pH-shifting, enzymatic hydrolysis, enzymatic cross-linking, complexation and modulation of amino acids. We base the discussion on diverse modified methods of nitrogen solubility index available to determine and analyze protein solubility followed by addressing how other indigenous components affect the solubility of plant proteins. Some nonproteinaceous constituents in proteins such as carbohydrates and polyphenols may exert positive or negative impact on protein solubility. Appropriate protein extraction and modification methods that meet consumer and manufacturers requirements concerning nutritious and eco-friendly foods with lower cost should be investigated and further explored.

Effect of High-Tunnel and Open-Field Production on the Yield, Cannabinoids, and Volatile Profiles in Industrial Hemp (Cannabis sativa L.) Inflorescence.

This study discovered the impact of high-tunnel (i.e., unheated greenhouse) and open-field production on two industrial hemp cultivars (SB1 and CJ2) over their yield parameters, cannabinoid development, and volatile profiles. Development of neutral cannabinoids (CBD, THC, and CBC), acidic cannabinoids (CBDA, THCA, and CBCA), and total cannabinoids during floral maturation were investigated. The volatile profiles of hemp flowers were holistically compared via HS-SPME-GC/MS. Findings indicated a high tunnel as an efficient practice for achieving greater total weight, stem number, and caliper, especially in the SB1 cultivar. Harvesting high-tunnel-grown SB1 cultivars during early flower maturation could obtain a high CBD yield while complying with THC regulations. Considering the volatile profiles, hemp flowers mainly consisted of mono- and sesquiterpenoids, as well as oxygenated mono- and sesquiterpenoids. Volatile analysis revealed the substantial impact of cultivars on the volatile profile compared to the production systems.

Investigating the impact of ultrasound on the structural, physicochemical, and emulsifying characteristics of Dioscorin: Insights from experimental data and molecular dynamics simulation.

This study investigated the impact of ultrasound treatment on dioscorin, the primary storage protein found in yam tubers. Three key factors, namely ultrasound power, duration, and frequency, were focused on. The research revealed that ultrasound-induced cavitation effects disrupted non-covalent bonds, resulting in a reduction in α-helix and ß-sheet contents, decreased thermal stability, and a decrease in the apparent hydrodynamic diameter (Dh) of dioscorin. Additionally, previously hidden amino acid groups within the molecule became exposed on its surface, resulting in increased surface hydrophobicity (Ho) and zeta-potential. Under specific ultrasound conditions (200 W, 25 kHz, 30 min), Dh decreased while Ho increased, facilitating the adsorption of dioscorin molecules onto the oil-water interface. Molecular dynamics (MD) simulations showed that at lower frequencies and pressures, the structural flexibility of dioscorin's main chain atoms increased, leading to more significant fluctuations between amino acid residues. This transformation improved dioscorin's emulsifying properties and its oil-water interface affinity.

Nutraceutical nanoemulsions: influence of carrier oil composition (digestible versus indigestible oil) on ß-carotene bioavailability.

BACKGROUND: Carotenoids, such as ß-carotene, are widely used in foods and beverages as natural colorants and nutraceuticals. We investigated the influence of carrier oil composition (ratio of digestible to indigestible oil) on the physical stability, microstructure and bioaccessibility of ß-carotene nanoemulsions using a simulated gastrointestinal tract model. RESULTS: ß-Carotene nanoemulsions (d < 150 nm) were formed by high-pressure homogenization using sucrose monoester and lysolecithin as emulsifiers, and mixtures of corn oil (digestible) and lemon oil (indigestible) as the lipid phase. All of the nanoemulsions underwent extensive droplet aggregation under mouth, stomach and small intestine conditions. The extent of free fatty acid production in the small intestine increased as the amount of digestible oil in the droplets increased. The bioaccessibility of ß-carotene also increased with increasing digestible oil content, ranging from ∼5% for the pure lemon oil system to ∼76% for the pure corn oil system. This effect was attributed to the ability of mixed micelles formed from triglyceride digestion products (free fatty acids and monoglycerides) to solubilize ß-carotene. CONCLUSIONS: This study provides important information for developing effective delivery systems for lipophilic bioactive components in food and beverage applications.

Oleofoams stabilized by monoacylglycerides: Impact of chain length and concentration.

Oleofoams are plant oil based whipped systems which have drawn academic and industry attention in recent years. The aim of this study was to determine the effect of fatty acid chain length and monoacylglyceride (MAG) concentration on the performance and structural properties of MAG-based oleofoams. Four different MAGs (monolaurin, monomyrystin, monopalmitin, and monostearin) were studied at three concentration levels (5, 10, and 15 wt%). The fatty acid chain length had a statistically significant impact on the size and shape of crystals formed, while higher MAG concentrations led to higher numbers of crystals in the continuous oil phase. These differences affected the performance and physical properties of the oleofoams: compared to other MAGs, monostearin based oleofoams were harder and exhibited higher values of G' and G″, had higher overrun and showed better stability. Lastly, through microscopy techniques it was successfully proved that monostearin-based oleofoams are stabilized by both bulk and Pickering stabilization.

Structural, functional properties, and volatile profile of hemp protein isolate as affected by extraction method: Alkaline extraction-isoelectric precipitation vs salt extraction.

The utilization of industrial hemp in food is gaining popularity. This study aimed to comprehensively investigate the impact of alkaline extraction-isoelectric precipitation (AE-IEP) and salt extraction (SE) dialysis on structural, functional properties and the volatile profile of hemp protein isolate (HPI). A higher protein content (97.21%) in SE extracted HPI (SE-HPI) was obtained than the AE-IEP extracted HPI (93.37%). In particular, protein subunit composition, structural properties were strongly influenced by the extraction methods. For example, SE-HPI exhibited a larger percentage of albumin (46.53%) and a lower amount of ß-sheet (52.65%) than its counterpart (albumin 20.92%, ß-sheet 54.46%). Consequently, SE-HPI showed the higher solubility, emulsion activity, and thermal stability. Interestingly, the volatile profile of the proteins showed that SE-HPI exhibited a lower number (21) of volatile compounds when compared to its counterpart (25). This study highlights the importance of establishing the relationships between extraction methods and functional attributes of HPI.

Lysozyme-phenolics bioconjugates with antioxidant and antibacterial bifunctionalities: Structural basis underlying the dual-function.

This work aims at adopting an Electron Paramagnetic Resonance (EPR) spectroscopic technique to help understanding protein-phenolic conjugation and final functionalities relationship as well as the underlying structural basis of antioxidant and antibacterial dual functionalities. Specifically, lysozyme (Lys) was conjugated with two natural phenolic acids, i.e. rosmarinic acid (RA) and gentisic acid (GA, our previous work) with obviously different molecular features. Lys-RA displayed 8.6- and 4.0-times enhanced antioxidant stoichiometry compared to the native Lys and ones with GA, respectively, due to the stronger antioxidant activity of RA. However, RA conjugation mitigated both enzymatic and antibacterial activities of Lys-RA conjugates. Such inhibition effect is attributed to the greater structural and surface property changes of Lys upon conjugating with RA. Furthermore, the polyphenol conjugation related structural basis of disturbance, reactivity and selectivity were explored via site-directed spin labeling (SDSL)-EPR. A dynamic picture of reactivity and selectivity of phenolics conjugation on Lys was proposed.

Antifungal activity, mycotoxin inhibitory efficacy, and mode of action of hop essential oil nanoemulsion against Fusarium graminearum.

This work aims to investigate antifungal, mycotoxin inhibitory efficacy of the hop essential oil (HEO) nanoemulsion and their mode of action (MOA) against Fusarium graminearum isolate, a fungal pathogen causing Fusarium Head Blight (FHB) in cereal crops. The HEO, primarily consisting of terpenes and terpenoids, was encapsulated in nanoemulsion droplets. Physically stable HEO-in-water nanoemulsion was fabricated using 0.5 wt% of tween 80 and 5 wt% oil phase comprising 30 % of Ostwald ripening inhibitor and 70 % of HEO. In terms of antifungal effect, HEO nanoemulsion could not only effectively inhibit mycelial growth and spore germination of F. graminearum isolates, but also remarkably suppress the production of deoxynivalenol (DON) and its derivatives in rice culture by applying 750 µg of HEO/g rice. Our studies on the MOA showed that HEO nanoemulsion could alter the contents of total lipid and chitin in outer cell membrane as well as damaging cytoplasmic membrane.

Molecular interaction mechanism and structure-activity relationships of protein-polyphenol complexes revealed by side-directed spin labeling-electron paramagnetic resonance (SDSL-EPR) spectroscopy.

Deciphering interactions between bioactive protein and polyphenols are critical for designing and controlling functional protein-polyphenol complexes. Herein, using the site-directed spin labeled T4 lysozyme (T4L) and rosmarinic acid (RA) as a model system, we combined electron paramagnetic resonance spectra to investigate molecular interaction mechanism of the protein-polyphenol complexes in structural or conformational details. Experimental results show that molecular interactions between T4L and RA are a process from order to disorder. TEM images display that the complexes finally assemble into quasi-spherical colloidal particles. When T4L/RA ratio is 1:1, the complexes exhibit the optimized enzymatic and antioxidant dual-functionalities due to the synergetic effect and protection mechanism. However, with excess addition of RA, the enzymatic and antioxidant activities of the complexes started to attenuate because the catalytic active site and bioactive hydroxyl groups were buried. The revealed high-resolution interaction process could help better understand the corresponding alterations between structure and functionalities.

Unraveling the role of germination days on the aroma variations of roasted barley malts via gas chromatography-mass spectrometry based untargeted and targeted flavoromics.

Roasting imparts malts with an increased amount of hedonic aromas. However, the relationship between the production of roasted malts and the generation of characteristic malt aromas remains unclear. In this study, roasted barley malts (RM) were prepared from three consecutive germination days (3, 4, 5D), and the aroma profiles among RM and base malt were holistically compared via HS-SPME-GC-MS/O-based flavoromics. Furthermore, the wort color, free amino acids, reducing sugars, and fatty acids compositions were determined before-and-after roasting. Results showed that roasting could flatten variations of precursors regardless of germination days. Additionally, based on quantitation of 53 aromas, a PLS-DA model was applied to differentiate all malts by 17 aromas with VIP ≥ 1. As for aroma harmony, RM with 4D-germination outstood due to a pleasant nutty note with the highest sweet-to-nutty index of 0.8. This work answers how germination days would impact the aroma of RM for the first time.

Effect of treatment of Fusarium head blight infected barley grains with hop essential oil nanoemulsion on the quality and safety of malted barley.

Fusarium mycotoxin contamination of malting barley has been a persistent food safety issue for malting companies. In this study, the effect of hop essential oil (HEO) nanoemulsion on fungal biomass and mycotoxin production during the malting process was evaluated. Furthermore, the localization of fungal hyphae on the surface and inside the tissue of barley and malts was observed. The application of HEO nanoemulsion reduced fungal biomass and deoxynivalenol (DON) contents at each stage of the malting process as compared to control. During malting process, the fungal hyphae on kernel surfaces was reduced appreciably after steeping. However, the increment of hyphae was observed between the husk and testa layer of barley after germination than raw barley grains. In addition to its antifungal activity, the antioxidant activity of HEO in the treated malts suppressed the formation of aldehydes. This study lays the foundation for the utilization of HEO in the malting industry.