Search Results (242)

Koshu wine, produced from the indigenous Japanese grape Vitis vinifera L. cv. Koshu exhibits a lower pH than other white wines, hindering malolactic fermentation (MLF) by lactic acid bacteria (LAB). Here, we aimed to isolate LAB strains capable of performing MLF under these challenging conditions to improve wine quality. Sixty-four Oenococcus oeni and one Lactobacillus hilgardii strain were isolated from Koshu grapes and wines that had undergone spontaneous MLF. MLF activity was assessed under varying pH, SO₂, and ethanol conditions in modified basal medium (BM) and Koshu model wine media. Expression of stress-related genes was analyzed using real-time PCR. Carbon source utilization was evaluated via API 50CH assays. All isolates degraded malic acid and produced lactic acid at 15 °C and pH 3.2 in BM without reducing sugars. Seven strains, all identified as O. oeni, demonstrated MLF activity at pH 3.0 in modified BM lacking added reducing sugars or tomato juice. Six wine-derived strains tolerated up to 12% ethanol, whereas the grape-derived strain was inhibited at 10%. In a synthetic Koshu wine model (13% ethanol, pH 3.0), wine-derived isolates exhibited higher MLF activity than commercial starter strains. In high-performing strains, mleA was upregulated, and most isolates preferred fructose, arabinose, and ribose over glucose. These findings suggest that indigenous O. oeni strains from Koshu wine possess unique stress tolerance and metabolic traits, making them promising candidates for region-specific MLF starter cultures that could enhance Koshu wine quality and terroir expression. Full article

Cordyceps takaomontana is a medicinal fungus with significant pharmacological value, but how soil microbes promote its growth remains unclear. We established a solid-state co-culture system involving C. takaomontana synnemata and its native soil fungi of Fusarium paeoniae and Bjerkandera minispora. Both F. paeoniae and B. minispora significantly promoted synnematal growth and enhanced antioxidant enzyme activities. Total triterpenoid content increased substantially. F. paeoniae markedly elevated levels of ergosterol peroxide, whereas B. minispora boosted accumulation of L-arabinose, ergotamine, and euphol. Metabolomics revealed that both fungi activated key metabolic pathways (including ABC transporters, mineral absorption, and protein digestion/absorption). F. paeoniae uniquely upregulated phenylalanine metabolism. This work elucidates the metabolic mechanisms underlying growth promotion of C. takaomontana mediated by F. paeoniae and B. minispora as well as deciphers potential pharmacologically active metabolites. These findings provide a foundation for strategically improving artificial cultivation and developing functional microbial inoculants. Full article

Environmental conditions, including nutrient composition and temperature, influence biofilm formation and antibiotic resistance in Escherichia coli. Understanding how specific metabolites modulate these processes is critical for improving antimicrobial strategies. Here, we investigated the growth and composition of Escherichia coli in both planktonic and biofilm states in the presence of L-arabinose, with and without exposure to the fluoroquinolone antibiotic levofloxacin, at two temperatures: 28 and 37 °C. At both temperatures, L-arabinose increased the growth rate of planktonic E. coli but resulted in reduced total growth; concurrently, it enhanced biofilm growth at 37 °C. L-arabinose reduced the efficacy of levofloxacin and promoted growth in sub-minimum inhibitory concentrations (25 ng/mL). Transcriptomic analyses provided insight into the molecular basis of arabinose-mediated reduced susceptibility of E. coli to levofloxacin. We found that L-arabinose had a temperature- and state-dependent impact on the transcriptome. Using gene ontology overrepresentation analyses, we found that L-arabinose modulated the expression of many critical antibiotic resistance genes, including efflux pumps (ydeA, mdtH, mdtM), transporters (proVWX), and biofilm-related genes for external structures like pili (fimA) and curli (csgA, csgB). This study demonstrates a previously uncharacterized role for L-arabinose in modulating antibiotic resistance and biofilm-associated gene expression in E. coli and provides a foundation for additional exploration of sugar-mediated antibiotic sensitivity in bacterial biofilms. Full article

This study aimed to optimize the enzyme-assisted extraction of polysaccharides (RTFPs) from Rosa roxburghii fruit using response surface methodology. Under the optimal extraction conditions, the yield of RTFPs reached 14.02%, which was close to the predicted value of 13.96%. The primary structural characteristics and the antioxidative and immunomodulatory activities of RTFPs were also examined. Structural characterization revealed that RTFPs comprise 36.38% neutral sugar, 48.83% uronic acid, and 7.29% protein. Their heteropolysaccharide structure features two distinct molecular weight fractions (1.87 × 10⁵ Da and 4.75 × 10³ Da) and a monosaccharide composition dominated by glucose (38.93%), arabinose (20.66%), galactose (20.58%), galacturonic acid (10.94%), and xylose (6.52%). Antioxidant assays demonstrated potent radical scavenging activity, with IC₅₀ values of 11 μg/mL (DPPH) and 150 μg/mL (ABTS), comparable to conventional antioxidants. Immunomodulatory studies on RAW264.7 macrophages revealed that RTFPs (100–400 μg/mL) significantly enhanced phagocytosis by 12.61–76.63% and stimulated the secretion of nitric oxide (NO) and tumor necrosis factor-α (TNF-α). These bioactivities are attributed to RTFPs’ high uronic acid content, moderate molecular weight distribution, unique monosaccharide profile, and highly branched conformation. Full article

E. coli attaches to, and forms biofilms on various surfaces, including latex and polystyrene, contributing to nosocomial spread. E. coli responds to both exogenous and endogenous insulin, which induces behavioral changes. Human insulin, a quorum signal surrogate for microbial insulin, may affect the ability of E. coli to interact with latex and polystyrene in the presence of various sugars. E. coli ATCC 25923 was grown in peptone (1%) yeast nitrogen base broth to either the logarithmic or stationary growth phase. Adherence to latex was determined using 6 × 6 mm latex squares placed in a suspension of washed cells (10³ CFU/mL; 30 min; 37 °C) in buffer containing insulin at 2, 20, and 200 µU/mL (Humulin^® R; Lilly) with and without mannose, galactose, fructose, sorbose, arabinose, xylose, lactose, maltose, melibiose, glucose-6-phosphate, glucose-1-phosphate, and glucosamine at concentrations reported to affect behavioral response. Attachment levels to latex were determined by the press plate method. Biofilm levels were measured in a similar fashion but with overnight cultures in flat bottom uncoated polystyrene plates. Controls were media, insulin, sugar, or buffer alone. Glucose served as the positive control. Overall, the stationary phase cells’ adherence to latex was greater, regardless of the test condition, than was measured for the logarithmic phase cells. The effect of insulin on adherence to latex was insulin and sugar concentration dependent. The addition of insulin (200 µU/mL) resulted in a significantly (p < 0.05) increased adherence to latex and biofilm formation on polystyrene compared with sugar alone for 12 of the 13 sugars tested with stationary phase bacteria and 10 of the 13 sugars tested with logarithmic phase bacteria. Adherence in response to sorbose was the only sugar tested that was unaffected by insulin. These findings show that insulin enhances E. coli’s association with materials in common usage in medical environments in a nutrition-dependent manner. Full article

One of the world’s most sustainable solutions is to replace fossil-based polymers with biopolymers. The production of xanthan gum can be optimized using various renewable and cost-effective raw materials, which is a key focus in industrial biotechnology. Xanthan gum is a bioengineered thickening, stabilizing, and emulsifying agent. It has unique properties for use in many industries (food, biotechnology, petrochemicals, agricultural, cosmetics, wastewater treatment) and medical applications. It is tasteless, environmentally safe, non-toxic, and biodegradable. The biotechnological production of xanthan gum depends on several factors: bacterial strain development, culture medium preparation, carbon sources, fermentation parameters and modes, pH, temperature, recovery, purification, and quality control regulations. Bio-innovative strategies have been developed to optimize the production of xanthan gum. A variety of carbon and nitrogen sources, as well as alternative renewable sources, have been used in the production of xanthan gum. The aim of the present study was to optimize the xanthan gum yield using Xanthomonas campestris bacteria and different carbon (D-glucose, D-sorbitol, lactose, sucrose, D-mannitol, D-fructose, erythritol, coconut palm sugar, L-arabinose, unrefined cane sugar), various nitrogen (bacterial peptone, casein peptone, L-glutamic acid, L-arginine, L-methionine, L-tryptophan, malt extract, meat extract, L-phenylalanine, soy peptone) and alternative carbon (orange peels, tangerine peels, lemon peels, avocado peels, melon peels, apple peels, cellulose, xylose, xylitol) sources. The xanthan gum samples were analyzed using antioxidant methods. Our study showed that using L-glutamic acid as the carbon source for 72 h of bacterial fermentation of Xanthomonas campestris resulted in the highest xanthan gum yield: 32.34 g/L. However, using renewable resources, we achieved a very high concentration of xanthan gum in just 24 h of fermentation. According to the reducing power and DPPH methods, the highest antioxidant activities were measured for xanthan gum whose biosynthesis was based on renewable resources. Xanthan gum structures have been verified by FT-IR and ¹H NMR analysis. The sustainable biotechnology study has the advantage of increasing the sustainable production of xanthan gum by using renewable alternative resources compared to other production processes. Xanthan gum continues to be a valuable biopolymer with a wide range of industrial applications while promoting environmentally friendly production practices. Full article

Wild fruits are distributed worldwide, but are consumed mainly in developing countries, where they are an important part of the diet. Still, in many other countries, they are consumed only locally. Blackthorn (Prunus spinosa L.) is an underutilized species rich in fibres and phenolic compounds, making it suitable as a potential functional food for supporting human health. Cold (Cw) and hot (Hw) water-extracted (poly)phenolic polysaccharide–protein complexes, differing in carbohydrate, phenolic and protein contents, were isolated from blackthorn fruits and characterized. The complexes exhibited molecular weights of 235,200 g/mol (Cw) and 218,400 g/mol (Hw), and were rich in pectic polymers containing galacturonic acid, arabinose, galactose and rhamnose, indicating a dominance of homogalacturonan (HG) [→4)-α-D-GalA(1→4)-α-D-GalA(1→]n and a low content of RGI [→2)-α-L-Rha(1→4)-α-D-GalA(1→2)-α-L-Rha(1→]n sequences associated with arabinan or arabinogalactan. Minor content of glucan, probably starch-derived, was also solubilized. Pectic polysaccharides were highly esterified and partly acetylated. Pharmacological testing was performed in male Dunkin–Hartley guinea pigs, a model with human-like airway reflexes. Both complexes affected airway defense mechanisms. Particularly, Hw significantly suppressed citric acid-induced cough, similar to codeine, and reduced bronchoconstriction comparably to salbutamol in a dose-dependent manner. These findings support further exploration of Hw as a natural antitussive and bronchodilatory agent. Full article

This study optimized ultrasound-assisted extraction (UAE) for polysaccharide isolation from Brassica rapa L. using Box–Behnken design, achieving a maximum yield of 41.12% under conditions of 60 °C, 60 min, 175 W ultrasonic power, and 30 mL/g liquid–solid ratios. The crude polysaccharide (BRAP) was purified via DEAE-52 cellulose and Sephadex G-100 chromatography, yielding BRAP1-1 with the highest recovery rate. Structural analyses (FT-IR, HPGPC, SEM, SEC-MALLS-RI) identified BRAP1-1 as a β-glycosidic pyranose polysaccharide (32.55 kDa) composed of fucose, rhamnose, arabinose, galactose, and galacturonic acid (molar ratio 0.81:4.30:3.61:1.69:89.59). In a humanized mouse model via fecal microbiota transplantation (FMT), BRAP1-1 significantly increased α-diversity indices (ACE, Chao1; p < 0.05) and altered β-diversity, with PCA explaining 73% variance (PC1: 60.70%, PC2: 13.53%). BRAP1-1 elevated beneficial genera (Lysinibacillus, Solibacillus, Bacteroides, etc.) while suppressing pathogens (Treponema, Flavobacterium, etc.). Six genera, including [Eubacterium]_coprostanoligenes_group and Bacteroidales (p < 0.05), correlated with acetic/propionic acid production. These findings demonstrate BRAP1-1’s potential to modulate gut microbiota composition and enhance intestinal homeostasis. Full article

To address the underutilization of Schizochytrium limacinum meal, polysaccharides from Schizochytrium limacinum meal (SLMPs) were prepared via ultrasonic-assisted eutectic-solvent-based extraction. Although polysaccharides exhibit promising application potential, the structural ambiguity of SLMPs necessitates systematic investigation to elucidate their structure–activity relationships, thereby providing a scientific foundation for their subsequent development and utilization. Using response-surface methodology (RSM), the optimized extraction conditions were determined as follows: ultrasonic temperature of 52 °C, ultrasonic duration of 31 min, ultrasonic power of 57 W, water content of 29%, and a material-to-liquid ratio of 1:36 g/mL. Under these conditions, the maximum polysaccharide yield reached 9.25%, demonstrating a significant advantage over the conventional water extraction method (4.18% yield). Following purification, the antioxidant activity and structural characteristics of SLMPs were analyzed. The molecular weight, monosaccharide composition, reducing groups, and higher-order conformation were systematically correlated with antioxidant activity. Fourier-transform infrared spectroscopy (FTIR), monosaccharide composition analysis, and ¹H nuclear magnetic resonance (NMR) spectroscopy revealed characteristic polysaccharide functional groups (C–O, O–H, and C=O). Monosaccharides such as glucose (Glc), galactose (Gal), and arabinose (Ara) were found to enhance antioxidant activity. High-performance gel permeation chromatography (HPGPC) indicated a molecular weight of 20.7 kDa for SLMPs, with low-molecular-weight fractions exhibiting superior antioxidant activity. Scanning electron microscopy (SEM) further demonstrated that ultrasonically extracted polysaccharides possess porous structures capable of chelating redox-active functional groups. These findings suggest that ultrasonic-assisted eutectic-solvent-based extraction is an efficient method for polysaccharide extraction while preserving antioxidant properties. Full article

As global milk production continues to rise, the disposal of expired milk contributes to environmental pollution and valuable resource wastage. This study presents the development of a novel L-arabinose isomerase, designated BmAIase12, and its application in the enzymatic recycling of expired milk. BmAIase12 exhibited a specific activity of 10.7 U/mg and showed optimal performance at 50 °C and pH 7.0. Furthermore, it exhibited higher activity than most other L-arabinose isomerases. It converted D-galactose into D-tagatose with a high conversion ratio of 53.3% after 48 h at 50 °C. The conversion efficiency of expired milk to D-tagatose was recorded at 40.62%, resulting in a maximum tagatose yield of 1.625 g/L. This was accomplished through the incorporation of β-galactosidase (120 U/mL) and Saccharomyces cerevisiae (30 mg/mL) to hydrolyze lactose and metabolize glucose, followed by the addition of 3 U/mL of BmAIase12. Ultimately, following purification, the purity of tagatose was determined to be 98%, and the final yield was 29.8%. These results suggest that BmAIase12 may serve as a promising enzyme for D-tagatose production due to its high conversion yield. Full article

Grapes are commonly processed into shelf-stable products such as raisins, wine, juice, and syrup-canned syrup goods. During processing, byproducts like skins and seeds are generated, which contain bioactive compounds including polysaccharides and polyphenols that exhibit diverse biological activities. The objective of this work was to thoroughly evaluate the impact of ultrasound technology on both the extraction efficiency and in vitro hypoglycemic activity of the polysaccharides derived from grape skin. The isolation and purification of the polysaccharides were carried out using chromatographic column techniques, and the monosaccharide components were determined through HPLC. The hypoglycemic activity of the polysaccharides from grape skin in vitro was analyzed in vitro considering their inhibitory effects on α-amylase and α-glucosidase. The polysaccharides from grape skins were extracted via an ultrasound-assisted methodology (under the following conditions: 50 °C, 50 min, 20 mL/g ratio, and 210 W), resulting in an 11.82% extraction yield of GSPs. Monosaccharide constituent analysis revealed that GSP-1-1 consisted of galacturonic acid, arabinose, rhamnose, galactose, glucose, glucuronic acid, mannose, and xylose in a molar ratio of 40.26:26.99:13.58:12.2:2.24:1.97:1.63:1.42. In vitro evaluations indicated that both GSP and GSP-1-1 exhibited notable suppression of α-amylase and α-glucosidase activities, two key enzymes in carbohydrate digestion. This dual inhibitory action positions these compounds as potential therapeutic agents for blood glucose management strategies. This work provides a new direction for addressing the byproducts of the grape canning industry and also offers a theoretical basis for the development of functional grape products. Full article

Hydroxyproline O-arabinosyltransferase (HPAT), a critical enzyme in plant glycosylation pathways, catalyzes the transfer of arabinose to the hydroxyl group of hydroxyproline residues. This enzyme contains a canonical GT95 glycosyltransferase, a structural hallmark of this carbohydrate-active enzyme family. HPAT mediates arabinosylation of diverse cellular targets, including cell wall extension and small signaling peptides. Emerging evidence has shown that HPAT orthologs regulate plant development and symbiotic interactions through post-translational modification of CLV1/LRR Extracellular (CLE) peptides. Although the molecular functions of HPAT genes have been characterized in model plants such as Arabidopsis thaliana and Lotus japonicus, their roles remain unexplored in Zea mays L. In this study, we used ZmHPAT2 homozygous mutants to explore the function of the maize HPAT gene. Sequence analysis identified a N-terminal signal peptide targeting the Golgi apparatus and promoter elements responsive to AM fungal colonization. Phenotypic analysis revealed its negative regulatory role: zmhpat2 promotes vegetative growth (increased plant height and accelerated flowering) and enhances AM symbiosis (increased colonization rate). Mechanistic studies demonstrated that ZmHPAT2 possesses dual regulatory functions—the activation of auxin signaling and repression of ZmMYB1-mediated arbuscular degradation pathways. In addition, overexpression of ZmHPAT2 in Lotus japonicus inhibits growth (reduced plant height) and impairs symbiotic interactions. Our findings establish ZmHPAT2 as a critical node to regulate auxin and symbiotic signaling, providing novel insights into plant glycosylation-mediated development. This work not only advances our understanding of maize growth regulation but also identifies potential targets for crop improvement through arabinosylation pathway manipulation. Full article

Microbially synthesized postbiotics have unique properties and advantages; however, systematic studies on the efficient production and biological functions of postbiotics from Bacillus subtilis are limited, which greatly restricts their applications. In this study, we obtained a novel crude exopolysaccharide (EPS) postbiotic from Bacillus subtilis H4. We systematically optimized the EPS production strategy using single-factor analysis, Plackett–Burman design, the path of steepest ascent method, and response surface methodology. The optimized EPS yield was significantly improved, with a maximum yield of 15.01 g/L under the addition of 4.12% soy peptone, 8.99% sucrose, and 0.06% MnSO₄. We found that EPS is a neutral, heterogeneous polysaccharide with a pyranose ring, with a molecular weight of 44,304.913 kDa and a melting point of 218 °C. It consists of glucose, galactose, arabinose, glucosamine, and mannose at a molar ratio of 58.85:19.81:14.75:10.89:6.58. EPS exhibits strong antioxidant capacities, scavenging ABTS and DPPH radicals with IC50 values of 1 and 6 mg/mL, respectively. Moreover, it shows notable anti-inflammatory properties, dramatically inhibiting the lipopolysaccharide (LPS)-induced elevation of nitric oxide (NO) levels and over-activation of the TLR4-NF-κB signaling pathway. These findings highlight the potential of EPS as a multifunctional bioactive compound, offering great promise for its application in the food, clinical, and feed industries. Full article

The electrocatalytic reduction of O₂ via two-electron reaction (2e-ORR) to H₂O₂ represents a promising alternative to the current anthraquinone process, since it is advantageous in the sustainable and decentralized production of H₂O₂. Herein, we report the development of oxygen and nitrogen-rich few-layered graphene-like materials (ms-dcda) by the one-step carbonization of biomass-sourced monosaccharides (D-glucose, D-fructose, D-galactose, D-ribose, D-xylose, L-arabinose, and D-mannose) with the aid of dicyandiamide for electrochemical O₂ reduction to H₂O₂. The ms-dcda materials were porous and possessed wrinkled morphology typical of graphene nanosheets. In H₂O₂ production via 2e-ORR in an acidic electrolyte, these ms-dcda materials were all active and stable catalysts, among which glu-dcda derived from D-glucose and dicyandiamide displayed the lowest onset potential of 0.553 V and the highest selectivity of up to 91.6%. The catalyst was also highly stable in chronoamperometric tests. Selective chemical titration of the C–OH and C=O groups revealed that the latter is far more active and selective than the former in 2e-ORR. Moreover, a positive correlation between the contents of C=O and pyrrolic N and the H₂O₂ partial current suggests that the pyrrolic N group also contributes to 2e-ORR. This work affords a facile strategy for the sustainable fabrication of metal-free carbon-based catalysts efficient for H₂O₂ electrosynthesis. Full article

D-tagatose is a functional sweetener with glucose-regulating and prebiotic properties, but its bioproduction from D-galactose faces many limitations, particularly the high production costs. In particular, the current biosynthesis of D-tagatose suffers from thermal instability and the substrate selectivity issues of L-arabinose isomerase (L-AI) required to convert D-galactose into D-tagatose. In this study, recombinant Escherichia coli BW25113/pQE-80L-araA_F118M/F279I expressing double mutant L-AI was immobilized to enhance its stability and reusability. The optimal conditions for whole-cell catalysis were 60 °C, pH 6.5, 5 mM Mn²⁺, and 20 h, with a yield of 55.2 g/L of D-tagatose. Immobilization with 3% sodium alginate and 2% CaCl₂ retained 90% of the production efficiency displayed by free cells. Notably, the immobilized cells exhibited enhanced heat resistance (60–70 °C) and operational stability, retaining 76% activity after five cycles. The D-tagatose production was further increased to 129.43 g/L by increasing the substrate concentration to 250 g/L. Compared to free cells, immobilized cells retained 83.6% of the initial yield up to 10 batches. This study presents a cost-effective and sustainable method for the production of D-tagatose using optimized whole-cell catalysis through immobilization, which paves the way to solve industrial challenges such as thermal instability and low substrate efficiency. Full article