Food Chemistry

Milk fat globules (MFG) are complex structures that emulsify the fat in milk and exhibit various biological activities. Using docosahexaenoic acid (DHA) as a model polyunsaturated fatty acid, we demonstrate for the first time that a MFG-enriched fraction (isolated from commercial raw and unhomogenized bovine milk) catalyzes the oxidation of exogenous polyunsaturated fatty acid to its oxylipins. Static incubation of the MFG-enriched fraction (at 20% w/v in phosphate buffer and 5% v/v ethanol) with 150 {micro}M of DHA for 1 h resulted in the production of [~]13 pmol/mg cream of DHA-derived oxylipins in free and esterified (i.e., bound) forms. High enrichment in free DHA-oxylipins was observed, wherein incubation with DHA resulted in >80% of all free oxylipins being derived from DHA, in contrast to control samples in which only <8% of all free oxylipins were DHA derivatives. The most abundant products generated were 17-hydroxydocosahexaenoic acid and 19(20)-epoxydocosapentaenoic acid. Oxylipin generation was dependent on the structural integrity of MFG, with mechanical disruption (vortexing) impairing oxylipin synthesis more severely than thermal treatment. These findings suggest that MFGs harbor multiple metabolically active enzymes, including cytochrome P450, lipoxygenases, acyl-CoA synthetases, and acyltransferases, that act cooperatively to synthesize and esterify DHA-derived oxylipins. In summary, this study highlights the dual potential of MFGs to serve both as a food-grade biocatalyst for the synthesis of DHA-derived oxylipins and as carriers for these bioactive lipids. Future studies should evaluate the bioavailability and physiological relevance of these metabolites when consumed in the diet.

In vegetarian diets, phytate is known to disrupt the adsorption of minerals. Fortifying foods with phytase, a therapeutic enzyme known to mitigate phytate, might increase the uptake of important nutrients. Phytase is susceptible to environmental stress such as heat and acidic conditions encountered during food processing. Therefore, we developed and optimized a core-shell microparticle composed of a phytase-chitosan core and a shell consisting of cross-linked alginate-{kappa}-carrageenan. Ethanol was used to precipitate the microparticles, and the ethanol concentration was optimized along with the chitosan and phytase ratio and the alginate-carrageenan concentration, to form stable core-shell microparticles. The optimized core-shell microparticles have a loading capacity of 32.7% with a high encapsulation efficiency of 80.3% and uniform micro-size with a diameter of 3.2 {micro}m and a poly-dispersity index of 0.178. Loaded phytase retained 62.7% enzymatic activity after heat treatment and digestion conditions. These results indicate that core-shell microparticles are suitable for retaining enzyme activity within the food matrix under typical food processing conditions. HighlightsO_LIDevelopment of size-controlled core-shell microparticles to protect phytase C_LIO_LIPhytase-chitosan microparticles are surrounded by an alginate-{kappa}-carrageenan shell C_LIO_LIOptimization achieved 32.7% loading capacity with a uniform size of 3.2 {micro}m C_LIO_LICore-shell microparticles retained 62.7% enzyme activity after heat and digestion C_LIO_LIPhytase powder (2 mg) is required for a single maize meal C_LI

Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO), the most abundant protein on Earth, is an attractive and sustainable food ingredient owing to its favourable nutritional and techno-functional properties. Leafy vegetables are particularly rich sources of RuBisCO; however, large-scale vegetable production generates substantial quantities of residual biomass throughout agri-food supply chains. Drying is widely used to stabilise this biomass and facilitate storage, transport, and handling, yet most reported RuBisCO extraction methods have been developed for fresh material and are poorly suited to dried feedstocks. Here, we present a simple, scalable, and cost-effective process for the recovery of food-grade RuBisCO from dried leafy biomass. Using spinach, rocket, and kale as model systems, efficient protein extraction was achieved from both freshly dried leaves and commercially available leaf powders without the need for resource-intensive processing. Application of the method to spinach yielded approximately 75 mg of high-purity RuBisCO per 100 g fresh-leaf equivalent, corresponding to an extraction efficiency of [~]70%, which increased to [~]90% following supplementation with 20 mM CaCl2. The recovered protein fraction also exhibited favourable foaming capacity and foam stability, demonstrating its potential as a functional food ingredient. This work provides a practical route for the valorisation of dried vegetable residues and supports the development of circular, waste-to-value supply chains for sustainable plant protein production.

Cassava and cocoyam are major dietary staples in sub-Saharan Africa, commonly processed by natural fermentation before consumption. Although fermentation reduces antinutritional compounds and improves food quality, its molecular effects remain poorly characterized. We used untargeted mass spectrometry-based metabolomics with a computational annotation pipeline to compare fermentation-induced molecular remodeling in the two tubers, which showed distinct responses. In cassava, 718 of 773 significant features (92.9%) were depleted, indicating a predominantly catabolic process. In cocoyam, the response was more balanced, with 385 of 1,013 features (38.0%) enriched, including di- and tripeptides consistent with proteolytic processing. Class analysis, molecular networking, and pathway enrichment revealed tuber-specific signatures: cassava was dominated by purine metabolism, whereas cocoyam showed stronger enrichment of amino acid pathways. Cyanogenic glycoside-related features were depleted, consistent with detoxification. Biotransformation prediction also suggested putative fermentation products absent from current databases, highlighting the under-characterized chemistry of these tubers.

LC/MS-based food lipidomics provides detailed information on intact lipid species, but the resulting datasets are often difficult to translate into concepts directly useful for food quality, processing, nutritional profiling and authenticity assessment. Here, we present Lipid Food Profile (LFP), a module of the LipidOne platform designed to convert annotated LC/MS lipidomics data into interpretable food-relevant lipid indices. LFP applies an in silico hydrolysis strategy to reconstruct acyl, alkyl and alkenyl chains from intact lipid species while preserving their lipid-class origin. The reconstructed information is then summarized into index categories related to food lipid quality, compositional balance, omega balance, oxidative stability, chain remodelling and ether-linked chain contribution. The interpretative value of LFP was evaluated using three published food lipidomics datasets addressing different analytical questions: X-ray-induced lipid remodelling in Chlorella vulgaris, spatial lipid heterogeneity in Mugil cephalus bottarga, and geographical-origin assessment of camel milk. Across these case studies, LFP recovered the main conclusions of the original lipidomics investigations, including treatment-associated lipid remodelling, inner-outer layer differences in bottarga and regional variation in camel milk. Importantly, LFP reorganized these findings into a smaller number of food-oriented indices, providing additional information on saturation balance, oxidative susceptibility, chain architecture and classification potential. Overall, LFP provides an interpretative layer for LC/MS food lipidomics that complement conventional fatty-acid analysis and molecular-species-based interpretation. By translating complex lipidomic tables into structured lipid index profiles, the module may support more accessible and chemically meaningful analysis of food composition, processing effects, lipid quality and exploratory traceability applications. LFP is freely accessible through the LipidOne web platform (LipidOne.eu). HighlightsO_LILipid Food Profile translates LC/MS food lipidomics into interpretable lipid indices. C_LIO_LIThe workflow preserves chain and lipid-class information without chemical hydrolysis. C_LIO_LIPublished case studies show that LFP recovers and extends previous interpretations. C_LIO_LILFP supports food quality, processing and exploratory origin/authenticity assessment. C_LIO_LIThe module complements conventional fatty-acid analysis and molecular lipidomics. C_LI

Botanical dietary supplements (like wheat, barley, teff, oats, white lupin, pumpkin seed, and chickpeas) may contain trace amounts of toxicants in addition to important micronutrients. Developing and validating a reliable protocol for the simultaneous quantification of Cu, Fe, Zn, Mo, Se, Mn, Pb, Al, Ni, and Cr using a PerkinElmer (NexIONTM2000 model) quadrupole ICP MS (including a He collision and reaction cell when needed) with closed vessel microwave digestion using (HNO3 + H2O2) was the aim of this study.The method was subsequently utilized in a sample survey, and the outcomes were evaluated against WHO/JECFA standards. From five study regions, twenty-seven farm-collected botanical powder samples representing seven species were acquired. To create one composite per species, field subsamples were cleaned, air dried, ground, and blended (nine subsamples per botanical: three grabs from each of three farms). HNO3/H2O2 was used to digest aliquots (0.250-0.500gm) in closed microwave containers. Internal standards, multi-point external calibration, procedural blanks, verified reference materials, matrix spikes, and duplicates were all used in ICP MSs multi-element quantitation. Method LODs/LOQs, accuracy (CRM recoveries), and precision (RSD) were calculated.The technique produced low LODs that were suitable for dietary evaluation (typical LOD ranges: Cu, Fe, Zn, Mn, Ni, Cr (0.001-0.01) mg/kg; Mo, Se, Pb, Al (0.002-0.05) mg/kg. For the majority of analytes, within-run RSDs were less than 5%, while CRM recoveries ranged from 88.9 to 110%. The concentrations of essential elements varied greatly (average mg/kg: Fe (280.7{+/-}25.6); Zn (6.0{+/-}0.541); Cu (2.8{+/-}0.269); Mn (398.3{+/-}23.8); {micro}gm/kg: Se (0.061{+/-}0.006); Mo (1.0 {+/-}0.022). Although some composites approached or exceeded conservative intake thresholds for Pb and Al under high consumption scenarios, toxic elements were generally low (mean mg/kg: Pb (0.062{+/-}0.007); Al(185.2{+/-}18.5); Ni(1.6{+/-}0.163); Cr(1.8{+/-}0.171).For the simultaneous nutritional and contaminant profiling of supplements derived from cereals and those not, the validated ICP- MS workflow with microwave HNO3 and H2O2 digestion is suitable. Accurate labeling and consumer safety can be supported by routine screening and supply chain controls.

Marine macroalgae are valuable sources of bioactive compounds. In this study, we thus investigated the chemical composition and biological activity of an extract from the green seaweed Ulva lacinulata, composed of small bioactive compounds. Comprehensive compositional analyses and high-resolution mass spectrometry revealed its diverse molecular profile composed in particular of peptides/amino acid derivatives, saccharides, low-chain fatty diacids, oxylipins and minerals. Its anti-inflammatory activity was assessed after 6 h pre-treatment in LPS-stimulated cultured RAW 264.7 macrophages, showing that it significantly and dose-dependently reduced the expression and/or secretion of pro-inflammatory cytokines such as TNF-alpha; and IL-6, and targeted the NF-kB signaling cascade. It modulated the SIRT1-AMPK signaling axis and increased the LC3-II/LC3-I ratio, supporting the activation of a controlled autophagic response. This work highlighted the potential of this marine-derived extract as a safe and effective functional ingredient for the development of functional food and/or dietary supplements targeting chronic low-grade inflammation.

Phycocyanin (PC) is the principal natural blue pigment used in functional beverages, but it rapidly loses color and aggregates under acidic conditions (pH {approx} 3). Experimental screening of stabilizers is costly and combinatorially intractable. Here we develop a chemoinformatics framework -- descriptor-based QSPR, a chemistry-prior heuristic, and virtual screening -- that learns from three rounds of commissioned screening (48 compounds, 6% hit rate) to predict stabilizer efficacy directly from molecular structure. In this genuinely small-data regime (3 positives), a LightGBM classifier built from 10 RDKit descriptors and 11 domain-expert charge/polymer features attained a leave-one-out AUC of 0.73, only marginally above a single-feature charge-density baseline (AUC 0.67); LOO sensitivity was 1/3 at threshold 0.5. A complementary chemistry-prior heuristic encoding anion-type priors substantially outperformed both, reaching AUC 0.95, indicating that explicit chemical knowledge captures information that descriptor-based ML cannot readily recover at this dataset size. SHAP analysis of the QSPR identified effective negative-charge density per unit, log molecular weight, polyphosphate identity, and functional-group density as the dominant features (jointly {approx}97% of mean |SHAP|), recovering the electrostatic-complexation mechanism without it being supplied as a prior. Virtual screening of 30 generally recognized as safe (GRAS) food additives nominated the pyrophosphate family -- led by sodium pyrophosphate decahydrate and sodium hexametaphosphate (SHMP), both at P {approx} 0.99 -- and the heuristic additionally flagged sodium phytate (IP{square}), which the descriptor model under-ranked at P = 0.045. Experimental validation at pH 3 and 46 {degrees}C for 7 days confirmed SHMP 2:1 (78.1 {+/-} 11.3% color retention), TSPP 2:1 (54.1 {+/-} 10.6%) and IP{square} 1:1 (52.7 {+/-} 9.0%), while a ternary IP{square} + STPP combination reached 83.8 {+/-} 11.9%, surpassing all single-component formulations. {zeta}-Potential measurements indicated a predominantly electrostatic origin for the protection (Pearson r = -0.82 between {zeta} and CR{square} {square} {square}; n = 24; p = 1 x 10{square} {square}). The framework, dataset and code are released to accelerate stabilizer discovery for other acid-sensitive food colorants and to provide a candid small-data benchmark.

Fungal contamination of food with yeast and moulds is associated with major economic losses due to spoilage and also poses health risks in the form of mycotoxin production. The strain Pantoea agglomerans APC 4211 isolated from leaves of Ilex aquifolium (holly tree) has broad spectrum antifungal activity against a variety of food spoilage fungi. Genomic analysis of the strain confirmed the presence of biosynthetic gene clusters potentially encoding for the enzymatic machinery required for the production of the antifungal lipopeptide herbicolin A. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis of the cell-free supernatant (CFS) confirmed the presence of molecular masses corresponding to herbicolin A (1300.8 Da), and herbicolin B (1138 Da). Purified herbicolin A has desirable properties for biotechnological applications, including potent antifungal activity against a range of spoilage fungi, thermal stability and resistance to proteases. Herbicolin A has low cytotoxicity against epithelial cell lines and has minimum inhibitory concentrations (MICs) lower than those of some commercial antifungal drugs (0.2 - 2.5 {micro}g/ml). In a model dairy system (10% skim milk), herbicolin A demonstrated excellent solubility and stability, effectively eliminating Aspergillus niger and Penicillium notatum at a concentration of 5 {micro}g/mL. In conclusion, herbicolin A is a potent, naturally occurring antifungal agent with the potential to be applied as a biopreservative in food systems, providing a safe, clean-label, and efficient compound for synthetic preservatives replacement. HighlightsO_LIHerbicolin A has a strong potential as a natural preservative for food protection C_LIO_LIHerbicolin A shows lower MICs than several antifungal agents C_LIO_LIHerbicolin A is stable under heat and resistant to proteolytic degradation C_LIO_LIHerbicolin A has strong solubility and stability in a model dairy system C_LIO_LIHerbicolin A indicates low cytotoxicity against epithelial cell lines C_LI Data summaryThe authors confirm all supporting data, code and protocols have been provided within the article or through supplementary data files.

Mushroom production generates large amounts of by-products, particularly stipes, which can represent up to half of the fruiting body biomass. Due to their similar composition to mushroom caps, these residues represent a promising substrate for the development of value-added foods. In this study, oyster mushroom stipes were used as a substrate for solid-state fermentation (SSF) with a Neurospora crassa strain isolated in Albacete to produce a novel meat analogue inspired by the oncom. Fermentation generated a cohesive matrix bound by hyphae that adopted the shape of the mold and exhibited a meat-like color, although with a softer texture. Nutritional analysis revealed a product with relatively low protein content but a complete amino acid profile, enriched in dietary fiber and containing unsaturated fatty acids. These results demonstrate that SSF with N. crassa provides a strategy to upcycle oyster mushroom by-products into fiber-rich meat analogues with potential applications in sustainable food systems.

Polyphenols are compounds synthesized by plants as part of their chemical defense system to counteract biotic and abiotic stressors. These compounds share two key chemical characteristics: their aromatic groups make them insoluble in water, while hydroxy groups provide redox properties. These characteristics may explain how polyphenols interact with mitochondrial membranes (which are lipophilic) and participate in redox (electron scavenging) reactions of the electron transport chain, ultimately affecting ATP synthesis via oxidative phosphorylation. This interaction accounts for both the beneficial and adverse effects of polyphenols. However, no research has examined how hydroxyl groups or a lipophilic environment influence the biological activity of polyphenols. Therefore, this study aimed to explore the impact of hydroxy groups and a lipophilic environment on the biological activity of polyphenols. We tested four polyphenols (quercetin, naringenin, resveratrol, and gallic acid) with varying numbers of hydroxyl and other functional groups to determine how hydroxyl groups affect their biological activity (toxicity) in Saccharomyces cerevisiae. Additionally, we evaluated different fatty acids to understand how a lipophilic environment influences polyphenol biological activity. The results of this study support the two main ideas of our hypothesis: 1) a lipid solvent increases the toxicity of polyphenols, and 2) the molecule with the most hydroxyl groups is the most toxic (as seen with quercetin, which has five hydroxyl groups). Consequently, the increased toxicity of polyphenols in lipid solvents, along with their association with oxidizable groups, opens the door to the development of new technologies based on polyphenols.

Glyceraldehyde (GA) contributes to the development of various diseases, such as diabetes and Alzheimer's disease via protein glycation and the formation of advanced glycation end products (AGEs); however, effective strategies for neutralizing GA are limited. Carnosine (Car), an imidazole dipeptide (IDP) that is abundant in meat, suppresses protein glycation by scavenging reactive aldehydes. There are only a few reports on the antiglycation activity of Car against GA. For other IDPs, such as anserine, balenine (Bal), and homocarnosine, there are almost no reports on their antiglycation activity. In this study, we demonstrated the antiglycation activity of four types of IDPs and 2-oxocarnosine (2-oxo-Car), an oxidized form of Car, against GA-induced intracellular protein glycation and neuronal cytotoxicity. Car and Bal exhibited significantly higher reactivity with GA compared with other IDPs and 2-oxo-Car. An in silico analysis suggested that the difference in reactivity is dependent upon intramolecular hydrogen bond formation and the conformation of each IDP. Although there were differences in reactivity with GA, LC-MS analysis revealed that all of the IDPs and 2-oxo-Car reacted with two molecules of GA to form adducts containing pyridinium rings. Car and Bal exhibited high reactivity with GA and markedly suppressed GA-induced cytotoxicity in SH-SY5Y cells. Western blot and qPCR analyses revealed that IDPs suppressed GA-induced protein glycation and the upregulation of endoplasmic reticulum and oxidative stress response genes. Our results indicate that IDPs represent a novel preventive approach to AGE-related diseases and provide a foundation for the development of strategies to treat GA-related neurotoxicity.

Sustainable agriculture has driven increased exploration of microbial inoculants as a promising strategy to boost plant growth for higher yield and enhance secondary metabolism to increase crop nutritional value. However, their influence on fruit-specific metabolites under field conditions remains understudied. This study investigated the impact of inoculating cranberry (Vaccinium macrocarpon) plants with Bacillus velezensis EB37 and Lachnum sp. EC5, applied individually and in combination, on cranberry fruit phytochemistry. Over two growing seasons (2019 and 2021), cranberries were collected from treated and control plots and analysed using untargeted and targeted LC-MS-based metabolomics. Multivariate analysis revealed no significant metabolomic differences due to treatments. However, samples clustered strongly by year of harvest, highlighting a pronounced environmental effect. Quantitative analysis of six representative phenolic compounds: chlorogenic acid, catechin, p-coumaric acid, phloridzin, myricetin, and quercetin, showed no statistically significant differences between treated and control cranberries. These findings indicate that microbial inoculation alone does not alter cranberry fruit metabolome, including phenolic levels, at field conditions. This study underscores how multiple factors, such as environmental conditions, can affect the outcome of microbial inoculation under field conditions and suggests that additional interventions may be required to achieve microbiome-based improvements in cranberry fruit quality.

Industrializing cultivated meat requires cell lines with high proliferative capacity, genetic stability, and suspension adaptability. We present a comprehensive multi-omics framework, integrating genomics, transcriptomics, and proteomics, to characterize a commercial duck Embryonic Stem Cell (dESC) line. Our analysis demonstrates continuous proliferation in protein-free suspension media while maintaining a stable genome and a functional conserved transcriptome. Broad-scale transcriptomics confirms the absence of hazardous pathway activation, and targeted assays verify sustained pluripotency marker expression during scale-up. Compositional analysis reveals a low-fat biomass containing all nine essential amino acids with an amino acid profile comparable to conventional duck meat. Furthermore, proteomic profiling demonstrates inter-batch reproducibility and protein distributions comparable to duck breast and liver. This study provides the first detailed molecular characterization of a commercial cultivated meat cell line, establishing a reference for the stability and safety assessment of future cultivated meat cell lines.

Glycation, a non-enzymatic reaction occurring between sugars and biological macromolecules, plays a critical role in ageing and disease pathogenesis. Methylglyoxal (MG) is a highly reactive -oxoaldehyde that leads to the formation of endogenous advanced glycation end products (AGEs). These AGEs are associated with diabetes and many other diseases, including neurodegeneration and cancer. This is often through interactions with the receptor for advanced glycation end products (RAGE). Inhibition of glycation/AGEs formation using natural products to target cancer is an area of recent interest. In vitro AGEs formation was observed by browning of samples, increased fluorescence, and carbonyl stress. MG induced changes in the structure of BSA were analysed using electrophoresis, spectroscopy, TEM, AFM, DLS, and CD spectroscopy. Our results show that AGEs form random structures, oligomeric aggregates, and {beta}-sheets. Thioflavin T and Congo red staining further validated these findings. Galangin and Caffeic acid demonstrated significant antiglycation activity, suppressing AGEs formation in vitro. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=134 SRC="FIGDIR/small/737425v1_ufig1.gif" ALT="Figure 1"> View larger version (39K): org.highwire.dtl.DTLVardef@113b391org.highwire.dtl.DTLVardef@7208a1org.highwire.dtl.DTLVardef@94c2e1org.highwire.dtl.DTLVardef@867b85_HPS_FORMAT_FIGEXP M_FIG C_FIG HighlightsO_LIMethylglyoxal-induced Advanced Glycation End Products were prepared in vitro C_LIO_LIMethylglyoxal -induced structural modifications in BSA C_LIO_LIAGEs were characterised using various parameters C_LIO_LIBoth fluorescent and non-fluorescent AGEs were formed. C_LIO_LIPhytochemical treatment induced inhibition of AGEs formation C_LI

In this research we inoculated Lacticaseibacillus casei (L. casei) into wheat sprouts and studied the viability of the bacteria in the sprout. L. casei (ATCC39392) strain was inoculated into sterilized wheat sprouts. The height of the sprouts and roots were checked and the active phenolic content, flavonoid compounds, and the antioxidant activity were measured. The bacterial viability was determined under simulated gastrointestinal (SGI) conditions. The physicochemical properties of the final product and its organoleptic properties were also investigated. The final confirmation of the presence of bacteria was also done by transmission electron microscope imaging. The number of bacteria increased from 8.18 Log CFU/g to 11.81 {+/-} 0.33 Log CFU/g. The increase in the phenolic content and antioxidant activity indicates the improvement of the nutritional value of the sprout. The physicochemical properties of the product changed due to the activity of bacteria. The inoculated bacteria also survived after exposure to SGI. The organoleptic properties of the product did not reveal a significant difference between the control and treatment groups. The increase in the number of bacteria and their survival after exposure to SGI indicates the suitable condition of wheat sprout as a proper substrate for L. casei bacteria.

Developing structured cultivated meat requires integrated solutions that combine scalable cell sources with edible, foodgrade materials capable of supporting highdensity growth and differentiation. Here, we evaluate bovine mesenchymal stem cells derived from embryonic stem cells (ESCderived iMSCs) as a scalable adipogenic cell source and develop an integrated workflow combining these cells with edible plantbased scaffolds for structured biomass generation. Cell identity and functionality were assessed using transcriptomic, morphological, gene expression, flow cytometric, and adipogenic differentiation analyses, in both adherent and suspension culture systems. In parallel, lentil, pea, and soy-based scaffold formulations were screened for cell attachment, proliferation, and biomass accumulation. Soybased scaffolds supported uniform cell distribution and robust growth and outperformed lentil-based scaffolds. Under dynamic culture conditions, bovine iMSCs cultured on soy-based scaffolds achieved highdensity growth, showing biomass accumulation (cell wet weight/scaffold wet weight) reached an average cell wet weight to scaffold wet weight ratio of 15% within three days. Cultures demonstrated active glucose metabolism and retained adipogenic differentiation capacity, confirmed by lipid accumulation and positive oil red O staining. These findings demonstrate an integrated cell-scaffold platform for rapid threedimensional biomass generation. This approach supports the development of a cell culture strategy for structured cultivated meat by combining defined cell sources with foodgrade scaffold technologies to improve scalability, structure, and nutritional relevance. HighlightsO_LIBovine ESC-derived iMSCs enable scalable adipogenic cell production C_LIO_LIEdible soy-based scaffolds support 3D attachment and biomass accumulation C_LIO_LIDynamic culture achieved [~]15% cell wet weight fraction within 3 days C_LIO_LIiMSCs retained adipogenic differentiation capacity on edible scaffolds C_LIO_LIIntegrated cell-scaffold culture supports structured cultivated meat prototypes C_LI

Cellobiose is an important disaccharide used in food, health, and biorefinery applications, but its efficient enzymatic production from crystalline cellulose remains challenging. In this study, crystalline cellulose was dissolved in ionic liquids and regenerated by dilution, and subsequently hydrolyzed at 80{degrees}C using a highly thermostable cellobiohydrolase, HmCel6A-3SNP. The enzyme retained activity in the presence of low concentrations of ionic liquids. Among the pretreatment conditions tested, cellulose treated with 1-butyl-3-methylimidazolium chloride ([Bmim]Cl) showed the highest enzymatic digestibility. After washing to remove residual ionic liquid, the reaction produced reducing sugars at levels 1.5-fold higher than those obtained in the presence of 10% [Bmim]Cl, with cellobiose accounting for approximately 96% of the products. Under the optimized conditions, the hydrolysis yield reached [~]36% after 48 hr. Structural analyses using birefringence imaging, electron microscopy, and Fourier transform infrared spectroscopy indicated that higher-order structural changes in regenerated cellulose strongly influence enzymatic accessibility. These results demonstrate the potential of combining ionic-liquid pretreatment with thermostable enzymes for selective cellobiose production from cellulose.

BackgroundControlled-environment cultivation of medicinal cannabis (Cannabis sativa L.) typically optimizes light conditions to enhance the biosynthesis of pharmaceutically important metabolites like cannabinoids. Such experimental strategies may also influence other specialized metabolites like terpenoids, flavonoids, alkaloids, among others. Previous untargeted metabolomics studies testing short wavelength conditions like UV and blue light have shown that terpenoids and prenylated flavonoids in cannabis leaves respond differentially. However, since metabolomic studies in cannabis have so far mostly focused on floral cannabinoids, a comprehensive untargeted study into cannabis floral metabolome response to short wavelengths is currently lacking. ObjectivesOur study investigates the impact of short wavelength usage on cannabis specialized metabolism, and in particular the influence of UVB, UVA, and blue light on the cannabis floral flavonoid metabolome and associated glycosylation moieties. MethodsCannabis plants were grown under a white background light and exposed to supplemental UVB, UVA, or blue light during the generative phase of the cultivation cycle. Treatments were compared to a reference white background light without UV or blue light. Metabolites from floral tissue were extracted and analyzed via ultra-performance liquid chromatography-tandem mass spectrometry. A comparative metabolomics workflow was designed and used to characterize the floral flavonoid metabolome and associated glycosylation moieties. ResultsOur results demonstrate how short wavelengths differentially affect the metabolism of natural product compound classes including polyketides and phenylpropanoids/shikimates. Blue light induced flavonoids similarly to how UVB did, while both UVA and blue light specifically induced flavanones accumulation. UVB showed the strongest regulatory effect on flavonoids production and glycosylation patterns. ConclusionsUVB reshapes the cannabis floral flavonoid metabolome by selectively stimulating the accumulation and structural modification of flavonoids. Therefore, UVB application in cannabis cultivation represents a useful horticultural strategy to increase inflorescence medicinal quality without affecting cannabinoid levels.

Raffinose family oligosaccharides (RFOs) are abundant in side streams from food and feed production from legumes, and the transition to plant-based diets increases the volume of such side streams. RFOs in the diet tend to have negative impacts on the consumers gut (e.g., nausea, bloating, diarrhoea), and in many ways, RFOs are comparable to lactose as a side stream from the dairy industry and symptoms associated with lactose intolerance. On the contrary, galactooligosaccharides (GOS) are recognized as prebiotics, and in this study we used a {beta}-galactosidase from Niallia circulans to produce potential prebiotics from RFOs (acceptors) and lactose (donor), which we hypothesized to have a lower fermentability than unmodified RFOs. The transglycosylation reactions resulted in RFO-based -{beta}-GOS, with NMR characterization showing ({beta}1-4) galactosylations on the non-reducing galactose end of RFOs as the major product. In reactions with RFOs, the characteristics were comparable to reactions with lactose alone and the new -{beta}-GOS products made up the largest fraction (by weight). A screening of 11 relevant gut and food microbe strains revealed that the gut commensal Bacteroides ovatus metabolised these modified oligosaccharides for growth whereas other strains grew only after adaption and others did not use them at all. This implies that mixed-linkage -{beta}-GOS are less fermentable by some microbes compared to raffinose, while other (beneficial) bacteria can still ferment them. The enzymatic synthesis established here is an interesting approach to upgrade abundant food side streams towards new prebiotics in a world where functional foods and food waste reduction receive increasing attention. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=77 SRC="FIGDIR/small/731070v1_ufig1.gif" ALT="Figure 1000"> View larger version (22K): org.highwire.dtl.DTLVardef@18e0e62org.highwire.dtl.DTLVardef@1525b4borg.highwire.dtl.DTLVardef@1e7be88org.highwire.dtl.DTLVardef@18df278_HPS_FORMAT_FIGEXP M_FIG C_FIG