Search Results (389)

Heme proteins are central to metabolism and stress responses but remain challenging to express recombinantly due to cytotoxicity and folding constraints. Phytoglobins (Pgbs) exemplify these difficulties, as expression protocols often fail to translate across protein species. Here, we used a cell-free protein synthesis (CFPS) platform powered by digital microfluidics to screen expression determinants for sugar beet Pgb 1.2 (BvPgb 1.2), its C86A variant, and three of eight newly identified oat Pgbs (AsPgbs), including their cysteine-to-alanine substituted variants. Benchmarking with multiple solubility tags and cell-free blends revealed protein- and variant-specific preferences, with alanine substitutions frequently improving expression and purification yields. Oxidative additives such as glutathione disulfide, alone or combined with protein disulfide isomerase, consistently enhanced production, underscoring the importance of redox environments for Pgb stability. Two selected variants were scaled up and yielded putative soluble apo-form proteins. The results highlight how CFPS enables rapid, parallelized identification of expression requirements while uncovering the role of conserved cysteines and redox conditions in Pgb biogenesis. Full article

This study designs stable fast-disintegrating oral granules (FDGs) of volatile liquid essential oils (cinnamon, lemon, rose, and peppermint oils) for olfactory rehabilitation. By adsorbing liquid-type essential oils onto colloidal silicon dioxide (1:1 w/w) and incorporating olive oil (1:1:0.25 w/w) as a volatile restrainer, the retention of major odorants—cinnamaldehyde, citral, phenethyl alcohol, and menthol—in cinnamon, lemon, rose, and peppermint oils, respectively, was markedly improved after 12 h of exposure at 40 °C. Then, free-flowing FDG system was formulated with sugar alcohols (mannitol, xylitol, and sorbitol), low substituted hydroxypropyl cellulose, and magnesium stearate, exhibiting rapid spreading and disintegration (<31.2 s) upon contact with aqueous media. The package of FDGs into four-layer laminated pouch afforded markedly prevent volatility of olfactory components, preserving >82% of cinnamaldehyde, citral, phenethyl alcohol, and menthol for 8 weeks under 25 °C/65% relative humidity (RH) conditions. In an in vitro volatilization test, major odorants were effectively volatilized from artificial saliva-wetted FDGs within 90 min depending on the volatility of each constituent. Therefore, this novel oral FDG system is expected to be a promising alternative for olfactory training for neurogenic smell dysfunction, providing enhanced storage stability, precise dosing, and patient compliance. Full article

This study investigated the effect of storage time on the quality of low-sugar apple jams partially substituted with steviol glycosides (SGs). Apple jams were prepared with 0%, 10%, 20%, 30%, and 40% sugar replacement using highly purified SGs (95.1%). The jams were evaluated immediately after production and after 3 and 6 months of storage at 22 °C in the dark. Physicochemical analyses included dry matter, total soluble solids, vitamin C, total ash, pH, titratable acidity, malic acid, and color parameters (L*, a*, b*). Sensory and microbiological assessments were also carried out. During storage, the dry matter content significantly decreased from 41.4% (control) to 35.6% (40% SGs), while titratable acidity increased from 10.69° to 16.73° (p < 0.05), and pH values remained stable (3.15–3.29). Vitamin C content decreased significantly (from 0.56 mg/100 g to 0.19 mg/100 g; 33–66% degradation). The color of jams became lighter with increasing SG substitution (L* increased from 17.19 to 24.73; ΔE up to 9.66) and slightly darkened after storage (ΔL ≈ −1.0). Microbiological analysis confirmed complete safety, with total colony counts < 10 CFU/g and no presence of Listeria monocytogenes or coagulase-positive Staphylococcus. Sensory evaluation by a trained panel (10 assessors, aged 34–56 years, with similar training in fruit and vegetable preserve evaluation) showed that jams with 10–30% SG substitution maintained desirable apple aroma and sweetness, whereas higher SG levels enhanced metallic odor (0.12–0.95 c.u.) and bitterness (0.2–1.9 c.u.) while slightly reducing apple flavor intensity (p < 0.05). Despite these differences, all jams remained acceptable after 6 months of storage. Overall, replacing up to 40% of sucrose with steviol glycosides provided microbiological stability, controlled color changes, and acceptable sensory quality, supporting the production of low-sugar jams in line with clean-label and sustainability trends in modern food technology. Full article

The rise in diabetes and obesity worldwide has created an urgent demand for low-sugar, nutrient-dense foods with appealing flavors. This study established an endogenous and “rapid validation–stable production” platform to enhance the flavor of edible tomato fruits by integrating two key technologies in the MicroTom cherry tomato: (1) TRV viral vector-mediated transient expression and (2) Agrobacterium-mediated stable genetic transformation. We employed the human sweet taste receptor TAS1R2 for in vitro functional validation and objectively demonstrated that tomato-derived recombinant thaumatin II exhibits receptor-binding activity equivalent to that of the native protein, overcoming the limitations of traditional sensory evaluation. Non-targeted metabolomic analysis (covering 1236 metabolites) confirmed that thaumatin II expression did not significantly alter the profiles of sugars, organic acids, or key flavor compounds in tomato fruits. This provides safety data supporting the development of “ready-to-eat sugar-substitute fruits.” Our strategy offers a solution and theoretical technical support for the development of low-sugar, high-nutrient foods. Full article

This study explored the utilization of date powder produced from non-marketable Sukkari and surplus dates as a natural sugar substitute in milk chocolate, with the dual objectives of improving product quality and reducing Saudi Arabia’s reliance on imported sugar. Molding chocolates were formulated with 10%, 20%, and 30% sugar substitution, while coating chocolates contained 20%, 40%, and 60%. Physical, chemical, and sensory properties were assessed immediately after production and following six months of refrigerated storage. Increasing substitution levels led to higher viscosity, with peak values observed at 30% in molding chocolate and 60% in coating chocolate. Ash and moisture contents also rose with substitution, whereas fat, protein, and carbohydrate levels remained largely unaffected. Energy content declined as substitution increased, and no significant differences were detected in peroxide or acid values between control and treated samples throughout storage. Sensory evaluations confirmed that molding chocolate with up to 20% substitution and coating chocolate with up to 40% substitution maintained comparable acceptability to control samples both initially and after storage. Overall, the findings recommend sugar replacement with date powder at 20% in molding chocolate and 40% in coating chocolate to achieve high-quality, nutritionally enhanced products with stable sensory and storage characteristics. Full article

Artificial sweeteners, or non-caloric sweeteners (NCSs), are widely consumed as sugar substitutes to reduce energy intake and manage obesity. Once considered inert, accumulating evidence now shows that NCSs interact with host physiology, altering gut microbiota composition and neural circuits that regulate feeding. This review synthesizes current knowledge on how NCSs disrupt the gut–brain axis (GBA), with particular focus on microbiota-mediated effects and neural reward processing. In homeostatic regulation, NCS-induced dysbiosis reduces beneficial taxa such as Akkermansia muciniphila and Faecalibacterium prausnitzii, diminishes short-chain fatty acid production, impairs gut barrier integrity, and promotes systemic inflammation. These changes blunt satiety signaling and favor appetite-promoting pathways. Beyond homeostasis, NCSs also rewire hedonic circuits: unlike caloric sugars, which couple sweet taste with caloric reinforcement to robustly activate dopaminergic and hypothalamic pathways, NCSs provide sensory sweetness without energy, weakening reward prediction error signaling and altering neuropeptidergic modulation by orexin, neurotensin, and oxytocin. Microbial disruption further exacerbates dopaminergic instability by reducing precursors and metabolites critical for reward regulation. Together, these top-down (neural) and bottom-up (microbial) mechanisms converge to foster maladaptive food seeking, metabolic dysregulation, and increased vulnerability to overeating. Identifying whether microbiome-targeted interventions can counteract these effects is a key research priority for mitigating the impact of NCSs on human health. Full article

Background: Artificial sweeteners, widely used as non-nutritive sugar substitutes, are increasingly prevalent in ultra-processed products. Although promoted for weight management due to their minimal caloric content, their impact on systemic inflammation remains uncertain. This systematic review of animal studies aims to evaluate the association between artificial sweetener consumption and inflammatory biomarkers. Methods: A systematic literature search was conducted up to May 2025 across PubMed, Web of Science, and Scopus, following PRISMA guidelines and registered in PROSPERO (CRD420251084004). Risk of bias was assessed using the ARRIVE guidelines and SCYRCLE’s risk of bias tool. Results: Thirty-seven animal studies were included: aspartame (n = 17), sucralose (n = 16), acesulfame potassium (n = 5), and saccharin (n = 4). Protocols varied in terms of dosage, exposure duration, animal models, and assessment of inflammatory outcomes, including C-reactive protein, interleukins (IL-6 and IL-1β), and tumor necrosis factor alpha. Aspartame and sucralose could elevate inflammatory markers, with sucralose also disrupting gut integrity and microbiota. Acesulfame K and saccharin showed variable, dose-dependent effects. Conclusions: This systematic review of animal studies suggests a possible mechanistic association between the consumption of certain artificial sweeteners and systemic inflammation. However, this relationship remains to be clarified and warrants exploration through well-designed, large-scale randomized controlled trials. Full article

Non-nutritive artificial sweeteners (NASs) are xenobiotics widely used in the food industry as sugar substitutes, since they provide few to no calories compared to sucrose. While NASs are considered safe at the acceptable daily intake (ADI) established by regulatory agencies, there is increasing controversy regarding their potential ability to promote metabolic derangements, especially to disrupt the gut microbiome balance. In this study, we analyzed a large cohort of the most commonly consumed beverages in Spain, categorizing them by the type of soda to determine the composition and content of the most frequently used NASs in the food industry. All commercial NAS formulations analyzed contained mixtures of different NASs. The NAS contents were always within regulated limits, although some samples yielded values close to these thresholds. Most soda samples analyzed contained NASs, even though the majority were not labeled as “zero sugars”, “no sugar added”, or “reduced calories”, which may mislead consumers. A preliminary statistical evaluation of the obtained results (cluster analysis) suggests that beverages can be grouped into three distinct clusters based on the total amount of NAS present in the samples. Differences in the total NAS content were significant among the three groups, with one cluster showing two- and four-fold higher levels than the others. Full article

Class 1 phytoglobins (Pgbs) are known for their multifunctional roles in plant stress responses, with recent studies suggesting broader interactions involving metabolic and transcriptional regulation. Interestingly, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) moonlights in many roles in colocalized spaces during cellular stress that are strikingly suitable for supporting Pgb function. This study investigates the molecular interactions of class 1 Pgb from sugar beet (Beta vulgaris), BvPgb 1.2, and an alanine-substituted mutant (C86A), focusing on their ability to bind GAPDH and DNA. Using dual-emission isothermal spectral shift (SpS) analysis, we report strong binding interactions with GAPDH, with dissociation constants (K_D) of 260 ± 50 nM for the recombinant wild-type protein (rWT) and a significantly stronger affinity for C86A (120 ± 40 nM), suggesting that the cysteine residue limits the interaction. Remarkably strong DNA-binding affinities were also observed for both variants, displaying biphasic binding. This behavior is characteristic of hexacoordinated globins and reflects the presence of two distinct species: a fast-reacting open pentacoordinated form and a slow-reacting closed hexacoordinated form with high apparent affinity. Here, the K_D in the open configuration was 120 ± 50 nm and 50 ± 20 nM for rWT and C86A, respectively. In the closed configuration, however, the cysteine appears to support the interaction, as the K_D was measured at 100 ± 10 pM and 230 ± 60 pM for rWT and C86A, respectively. Protein–protein docking studies reinforced these findings, revealing electrostatically driven interactions between BvPgb 1.2 and GAPDH, characterized by a substantial buried surface area indicative of a stable, biologically relevant complex. Protein–DNA docking similarly confirmed energetically favorable binding near the heme pocket without obstructing ligand accessibility. Together, these findings indicate a potential regulatory role for BvPgb 1.2 through its interaction with GAPDH and DNA. Full article

Non-centrifugal cane sugar (NCS) is an artisanal product, also known as rapadura or brown sugar, and it is consumed both as a dessert and as a substitute to refined sugar. Despite being a source of essential nutrients, inorganic contaminants may be found in rapadura composition. Thus, this study aimed to optimize and to apply a method for As, Cd, and Pb determination in 72 NCS samples commercialized in Latin America. The method consisted of acid extraction of the inorganic contaminants using an ultrasound bath, and the determination was conducted by inductively coupled plasma optical emission spectroscopy (ICP OES). The method optimization was performed using a 2² central composite design, considering time and oxidant mixture as key parameters, and the best conditions were verified by extracting the inorganic contaminants using a 15% oxidant mixture for 20 min. The acid extraction method using an ultrasound bath was considered adequate, with values for limits of detection and quantification between 0.005 and 0.039 mg kg⁻¹, respectively, and trueness (spiked experiments and certified reference material) ranging from 93 to 108% for all analytes. Rapadura samples from Latin America presented low levels for As and Cd, <0.012 µg kg⁻¹ and <0.005–0.045 mg kg⁻¹, respectively. For Pb, all samples presented quantifiable levels, and 33% were not within the requirements established by the Brazilian and The Southern Common Market (MERCOSUR) regulations. Thus, monitoring the levels of inorganic contaminants in non-centrifugal cane sugar is fundamental to provide safety for consumers. Full article

This manuscript provides an in-depth review of both artificial and natural sweeteners, including polyols and plant-derived alternatives, examining their sweetening potency, glycemic index, modes of action, and applications in the food sector. The rising demand for sugar substitutes, fueled by health concerns such as obesity and diabetes, has prompted significant research into low-calorie and non-nutritive sweeteners. This work categorizes sweeteners into synthetic options (such as aspartame, sucralose, saccharin) and naturally occurring ones (such as stevia, monk fruit, and polyols like sorbitol, xylitol, erythritol), focusing on physico-chemical characteristics, relative sweetness (ranging from 100 to 220,0000 times sweeter than sucrose), and glycemic index, important for their use in diabetes-friendly food products. The current manuscript examines how these sweeteners interact with taste receptors to induce sweetness perception without contributing significant calories. It also discusses their health implications and controversies and limitations regarding healthy and safety data, process feasibility, market application trends, environmental stability, and commercialization challenges. The review also addresses challenges in scaling production and ensuring the economic viability of plant-based sweeteners, offering a forward-looking perspective on their commercialization in the food industry. Full article

Extensive research is underway on a variety of functional foods that support consumer health. A promising combination is milk (a drink with high nutritional value), fortified with spices (naturally rich in antioxidant compounds) and supplemented with low-calorie, health-promoting sweeteners. The aim was to analyze the antioxidant activity (AA), total polyphenol content (TPC), and mineral composition of milk beverages fortified with a mixture of spices and sweeteners and to verify the interactions between these ingredients. Twenty-four variants of milk drinks were prepared with the addition of three types of spice mixtures (1:1) of clove (Cl), cinnamon (Ci), and turmeric (Tu) with the shares of 2.5%, 5%, 7.5%, and 10%, and two types of sweeteners—erythritol (E) and stevia (S)—as well as six control samples, including three 10% aqueous solutions of spice mixtures. AA was measured using the ABTS, FRAP, and DPPH methods. TPC was determined using the Folin–Ciocalteu method. Mineral content (Ca, Mg, Fe, Cu, and Zn) was assessed using the FAAS/FAES method. The highest AA was demonstrated for beverages with Cl-Ci and Cl-Tu, whereas the highest TPC was found in those with Cl-Tu. AA and TPC values increased with the increase in the share of spices in the beverages, and both measurements were significantly higher in beverages with erythritol compared to those with stevia. Despite the 4–5 times lower TPC, 3–8 times higher AA was demonstrated in beverages with the addition of 10% spice mixtures than their corresponding control samples (aqueous solutions with the addition of 10% spice mixtures), which suggests the great role of interactions between nutrients in food. For beverages with 10% Cl-Ci/Cl-Tu mixtures, significantly higher ABTS, FRAP, and DPPH were observed than would result from the simple sum of AA of the components (synergistic effect). The opposite (antagonistic) effect was observed for beverages with Ci-Tu. AA was positively correlated with the content of Fe, Mg, and Cu; negatively with Ca; and not correlated with Zn. The results suggest that the most health-promoting properties are exhibited by the milk drink with the addition of 10% Cl-Tu and erythritol, demonstrating the highest AA and TPC, the strongest synergistic effect of the components, and the highest content of Mg and Zn. This study highlights the importance of carefully selecting and combining ingredients to maximize the antioxidant properties of functional beverages. However, further research is needed to expand knowledge on this issue. Full article

I-motifs are non-canonical, cytosine-rich DNA structures stabilized by hemiprotonated C•C⁺ base pairs, whose formation is highly pH-dependent. While certain chemical modifications can enhance i-motif stability, modifications at the sugar moiety often disrupt essential inter-strand contacts. In this study, we examine the structural and thermodynamic impact of incorporating 2′-fluoro-ribocytidine (2′F-riboC) into i-motif-forming sequences derived from d(TCCCCC). Using a combination of UV, ¹H NMR, and ¹⁹F NMR spectroscopy, we demonstrate that full substitution with 2′F-riboC strongly destabilizes i-motif, whereas partial substitutions (one or two substitutions per strand) support well-folded structures at acidic pH (pH 5). High-resolution NMR structures reveal well-defined i-motif architectures with conserved C•C⁺ pairing and characteristic interstrand NOEs. Sugar conformational analysis reveals a predominant North pucker for cytosines, which directs the fluorine substituent toward the minor groove of the i-motif. ¹⁹F NMR further confirms slow exchange between folded and unfolded species, enabling the simultaneous detection of both under identical experimental conditions and, consequently, highlighting the utility of fluorine at the 2′ sugar position as a spectroscopic probe. These findings provide insights into fluorine-mediated modulation of i-motif stability and further extend the utility of ¹⁹F NMR in nucleic acid research. Full article

Low-calorie sweeteners (LCS) are widely utilized as sugar substitutes due to their intense sweetness, thermal stability, and applicability in weight management and diabetic-friendly products. However, increasing evidence has raised concerns about their potential long-term effects on metabolic health, glucose regulation, cardiovascular function, carcinogenicity, and gut microbiota composition. This review systematically evaluates the pharmacokinetics, metabolic effects, and associated health outcomes of major LCS. Mechanistically, LCS exert effects via sweet taste receptor-mediated pathways, altering glucose absorption, insulin secretion, and intracellular signaling cascades. Additionally, LCS influence gut microbiota composition, with certain agents promoting dysbiosis and glucose intolerance. While some findings support the metabolic benefits of selected LCS, others underscore potential risks, necessitating cautious interpretation. In conclusion, while LCS offer viable alternatives to sugar, their health effects are context-dependent and may vary across different sweeteners and populations. Long-term, high-quality clinical trials are essential to elucidate their safety and efficacy. Full article

The market for plant-based alternatives to animal foods has increased rapidly in the past decade, mainly due to consumer demand. Little evidence is available regarding nutritional impacts, drivers, and barriers to using these products as substitutes for animal foods in real-life conditions. This pilot study followed 16 Danish adults (30 ± 11 years old; 11 females) for 4 weeks with substituting milk, cheese, and yogurt with plant-based analogues to dairy (PBADs) and assessed their drivers and barriers to applying the intervention with a mixed-method approach. PBADs are constantly compared to their animal counterparts, both regarding product characteristics, such as price and sensory properties, as well as cultural roles and subjective memories. The mixed methods showed dairy attachment, price, and taste were the main barriers to consuming PBAD, while changes in life and social circles were drivers (qualitative data). As for the liking of PBADs, plant-based yoghurt was the preferred intervention product (73.5/100, p < 0.05), followed by plant-based drinks (65.9/100), while plant-based cheese was the lowest rated (47.9/100, p < 0.05). As for dietary changes, a lower average intake of sugars, saturated fatty acids, cholesterol, calcium, phosphorus, and zinc was observed after the intervention. Additionally, this study describes the attachment of the study population to milk and dairy products. It shows that choosing dairy is beyond nourishment but is connected to tradition, culture, pleasure, memories, and a sense of belonging. In contrast, there is no history or attachment to PBADs. Full article