Galactose Glycopolymer-Grafted Silica Nanoparticles: Synthesis and Binding Studies with Lectin.

Weak binding of carbohydrates with protein receptors possesses serious drawbacks in the advancement of therapeutics; however, the development of strategies for multipoint interactions between carbohydrates and protein can overcome these challenges. One such method is developed in this work where glycopolymer-grafted silica nanoparticles with a large number of carbohydrate units are prepared for the interactions with multiple binding sites of the protein. First, a glycomonomer, ß-d-galactose-hydroxyethyl methacrylate (ß-GEMA), was synthesized in a two-step process by coupling ß-d-galactose pentaacetate and hydroxyethyl methacrylate (HEMA), followed by deacetylation for the preparation of poly(ß-GEMA) glycopolymers (GPs). Further, the poly(ß-GEMA) chains were grafted onto the silica nanoparticle (SiNP) surface by utilizing the "grafting-from" strategy of surface-initiated reversible addition-fragmentation chain transfer (RAFT) polymerization to prepare p(ß-GEMA)-grafted SiNPs (GNPs). Five different chain lengths ranging from 10 to 40 kDa of the GPs and the GNPs were prepared, and various characterization techniques confirmed the formation of GPs and grafting of the GPs on the SiNP surface. The particle size of GNPs and the number of GPs grafted on the SiNP surface showed a strong dependence on the chain length of the GPs. Further, the GNPs were subjected to a binding study with ß-galactose-specific protein peanut agglutinin (PNA). A much stronger binding in the case of GNPs was observed with an association constant ∼320 times and ∼53 times than that of the monomeric methyl-ß-d-galactopyranoside and the GPs, respectively. Additionally, the binding of the PNA with GNPs and GPs was also studied with varying chain lengths to understand the effects of the chain length on the binding affinity. A clear increase in binding constants was observed in the case of GNPs with increasing chain length of grafted GPs, attributed to the enhanced enthalpic and entropic contributions. This work holds its uniqueness in these improved interactions between carbohydrates and proteins, which can be used for carbohydrate-based targeted therapeutics.

Atropisomerism Observed in Galactose-Based Monosaccharide Inhibitors of Galectin-3 Comprising 2-Methyl-4-phenyl-2,4-dihydro-3H-1,2,4-triazole-3-thione.

Galectin-3 (Gal-3) is a carbohydrate binding protein that has been implicated in the development and progression of fibrotic diseases. Proof-of-principal animal models have demonstrated that inhibition of Gal-3 is a potentially viable pathway for the treatment of fibrosis─with small molecule Gal-3 inhibitors advanced into clinical trials. We hereby report the discovery of novel galactose-based monosaccharide Gal-3 inhibitors comprising 2-methyl-4-phenyl-2,4-dihydro-3H-1,2,4-triazole-3-thione (compound 20) and 4-phenyl-4H-1,2,4-triazole (compound 15). Notably, hindered rotation caused by steric interaction between the 3-thione and ortho-trifluoromethyl group of compounds 20, 21 induced formation of thermodynamically stable atropisomers. Distinct X-ray cocrystal structures of 20 and 21 were obtained, which clearly demonstrated that the configuration of 21 proscribes a key halogen bonding σ-hole interaction of 3-chloro with carbonyl oxygen of Gly182, thereby leading to significant loss in potency. Ultimately, 20 and 15 were evaluated in mouse pharmacokinetic studies, and both compounds exhibited oral exposures suitable for further in vivo assessment.

The galactomannan-EGCG physical complex: Effect of branching degree and molecular weight on structural and physiological properties.

Polysaccharides and polyphenols are bioactive components that co-exist in many plant foods. Their binary interaction in terms of the structure-function relationships, however, has not been well clarified. This study elucidated the correlation between the structural and physiological properties of galactomannan (GM) -catechin monomer complexes and GM with different branching or molecular weight (Mw). Results indicated that locus bean gum with lower branching degree (Gal/Man is 0.259) bound more readily to EGCG with adsorption rate of 19.42 %. EGCG and ECG containing galloyl groups were more inclined to form hydrogen bonds with GMs, significantly improving the adsorption by GMs. The introduction of EGCG could enhance the antioxidant activity and starch digestion inhibition of GM, which positively correlated with the adsorption capacity of EGCG. The guar gum (GG) with higher Mw (7384.3 kDa) could transport 71.51 % EGCG into the colon, while the retention rate of EGCG reaching the colon alone was only 46.33 %. Conversely, GM-EGCG complex with lower Mw (6.9 kDa) could be readily utilized by gut microbiota, and increased production of short-chain fatty acids (SCFAs). This study elucidated the structure-properties relationship of GM-EGCG complexes, and provide a new idea for the development and precision nutrition of polysaccharides-polyphenol complexes fortified functional foods.

Non-carrier immobilization of yeast cells by genipin crosslinking for the synthesis of prebiotic galactooligosaccharides from plant-derived galactose.

Galactooligosaccharides (GOS), as mimics of human milk oligosaccharides, are important prebiotics for modulating the ecological balance of intestinal microbiota. A novel carrier-free cell immobilization method was established using genipin to cross-link Kluyveromyces lactis CGMCC 2.1494, which produced ß-galactosidase, an enzyme essential for GOS synthesis. The resulting immobilized cells were characterized as stable by thermogravimetric analysis and confirmed to be crosslinked through scanning electron microscopy analysis (SEM) and Fourier transform infrared spectroscopy (FTIR). The Km and Vmax values of ß-galactosidase in immobilized cells towards o-nitrophenyl ß-D-galactoside were determined to be 3.446 mM and 2210 µmol min-1 g-1, respectively. The enzyme in the immobilized showed higher thermal and organic solvent tolerance compared to that in free cells. The immobilized cells were subsequently employed for GOS synthesis using plant-derived galactose as the substrate. The synthetic reaction conditions were optimized through both single-factor experiments and response surface methodology, resulting in a high yield of 49.1 %. Moreover, the immobilized cells showed good reusability and could be reused for at least 20 batches of GOS synthesis, with the enzyme activity remaining above 70 % at 35 °C.

Computational study on the Maillard reactions of glucose and galactose with lysine.

CONTEXT: Milk has nutrient-rich but thermal sensitive matrix that undergoes varying degrees of Maillard reaction (MR) at heating conditions. The MR mainly occurs between lysine residues (Lys) and lactose composed of glucose (Glc) and galactose (Gal), which are abundantly sourced from dairy products. In the present study, the MRs of Glc and Gal with Lys at the initial and intermediate stages have been investigated theoretically using density functional theory (DFT) to simulate the gaseous and aqueous phases. Reaction mechanisms have been proposed, and relative energy changes of different steps were calculated according to the total mass balance. The calculations reveal that both Nα- and Nε-amine groups of Lys can react with the carbonyl functional group of Glc and Gal with the similar potential energy profiles, and Gal is more reactive than Glc. However, the barrier in Nε-channel is lower than in Nα-channel, indicating a faster reaction rate through the former channel compared with the latter. The 5-hydroxymethyl-2-furfural (HMF) and derivative are formed under 3-deoxysone route in the intermediate stage. The calculation results are helpful for proposing a reasonable MR mechanism and suggesting possible control methods of the MRs. METHODS: In this study, different levels of DFT calculations have been conducted to investigate the mechanisms and favorability of generating MR products in Glc-Lys and Gal-Lys models at initial and intermediate stages in the gaseous and aqueous conditions. In order to elucidate the molecular models from the perspectives of chemistry and geometry, DFT calculations were performed by the mean of B3LYP functional at basis sets of 6-311 + + G (d, p) and 6-311 + + G (2df, 2p) with optional solvation settings. To examine the solvation effect, the study further constructed models with solvent H2O and calculated in wB97XD functional with 6-31 + G (d) basis set. All computations were carried out Gaussian 09 suite of quantum chemistry software.

Doxorubicin-galactomannan nanoconjugates for potential cancer treatment.

In this study, we report the synthesis and characterization of pH-responsive nanoconjugates for targeted drug delivery. Galactomannan extracted from D. regia seeds was oxidized to form aldehyde groups, achieving a percentage of oxidation of 25.6 %. The resulting oxidized galactomannan (GMOX) was then copolymerized with PINIPAm-NH2, yielding a copolymer. The copolymer exhibited signals from both GMOX and PNIPAm-NH2 in its NMR spectrum, confirming successful copolymerization. Critical association concentration (CAC) studies revealed the formation of nanostructures, with lower CAC values observed at higher temperatures. The copolymer and GMOX reacted with doxorubicin (DOX), resulting in nanoconjugates with controlled drug release profiles, especially under acidic conditions similar to tumor microenvironments. Cytotoxicity assays demonstrated significant efficacy of the nanoconjugates against melanoma cells with reduced toxicity towards healthy cells. These findings underscore the potential of the pH-responsive nanoconjugates as promising candidates for targeted cancer therapy, offering improved therapeutic efficacy and reduced systemic side effects.

Co-immobilization of ß-agarase and α-agarase for degradation of agarose to prepare bioactive 3,6-anhydro-L-galactose.

Agarose from biomass can be used to synthesize the rare sugar 3,6-anhydro-L-galactose (L-AHG), and the new synthesis route and functional properties of L-AHG have always been the focus of research. Here we developed a novel method to co-immobilize Aga50D and BpGH117 onto streptavidin-coated magnetic nanoparticles and achieved the conversion of agarose to bioactive L-AHG in one pot. Results showed that enzymes were successfully immobilized on the carrier. The activity of co-immobilized enzymes was 2.5-fold higher than that of single immobilized enzymes. Compared with free enzymes, co-immobilized enzymes exhibited enhanced thermal stability. The co-immobilized enzymes retained 79.45 % relative activity at 40 °C for 3 h, while the free enzymes only possessed 21.40 % residual activity. After eight cycles, the co-immobilized enzymes still retained 73.47 % of the initial activity. After silica gel chromatography, the purity of L-AHG obtained by co-immobilized enzymes hydrolysis reached 83.02 %. Furthermore, bioactivity experiments demonstrated that L-AHG displayed better antioxidant and antibacterial effects than neoagarobiose. L-AHG had broad-spectrum antibacterial activity, while neoagarobiose and D-galactose did not show an obvious antibacterial effect. This study provides a feasible method for the production of L-AHG by a co-immobilized multi-enzyme system and confirms that L-AHG plays a key role in the bioactivity of neoagarobiose.

Poly(allylamine)-adorned heptylcarboxymethyl galactomannan nanocarriers of canagliflozin for controlling type-2 diabetes: Optimization by Box-Behnken design and in vivo performance.

In the past three decades, the prevalence of type-2 diabetes has arisen dramatically in countries of all income levels. A novel, most effective nanotechnology-based strategy may reduce the prevalence of diabetes. Recently, the shell-crosslinked polysaccharide-based micellar nanocarriers (MNCs) have shown great promise in terms of stability, controlled drug release, and improved in vivo performance. In this study, heptyl carboxymethyl guar gum was synthesized and characterized by ATR-FTIR, 1HNMR spectroscopy, surface charge, critical micelle concentration (23.9 µg/mL), and cytotoxicity analysis. Box-Behnken design was used to optimize the diameter, zeta potential, drug entrapment efficiency (DEE), and drug release characteristics of poly (allylamine)-crosslinked MNCs containing canagliflozin. The optimized MNCs revealed spherical morphology under TEM and had 149.3 nm diameter (PDI 21.2 %), +53.8 mV zeta potential, and 84 % DEE. The MNCs released about 63 % of the drug in 12 h under varying pH of the simulated gastrointestinal fluid. DSC and x-ray analyses suggested amorphous dispersion of drugs in the MNCs. CAM assay demonstrated the biocompatibility of the MNCs. The MNCs showed hemolysis of <1 %, 85 % mucin adsorption, and stability over three months. The MNCs demonstrated excellent anti-diabetic efficacy in streptozotocin-nicotinamide-induced diabetic rats, continuously lowering blood glucose levels up to 12 h.

Inequality relations for NMR-based polymer homoblock analysis and extended application: Reanalysis of historical data on alginates, chitosans, homogalacturonans, and galactomannans.

There has been a long-standing bottleneck in the quantitative analysis of the frequencies of homoblock polyads beyond triads using 1H and 13C NMR for linear polysaccharides, primarily because monosaccharides within a long homoblock share similar chemical environments due to identical neighboring units, resulting in indistinct NMR peaks. In this study, through rigorous mathematical induction, inequality relations were established that enabled the calculation of frequency ranges of homoblock polyads from historically reported NMR-derived frequency values of diads and/or triads of alginates, chitosans, homogalacturonans, and galactomannans. The calculated homoblock frequency ranges were then applied to evaluate three chain growth statistical models, including the Bernoulli chain, first-order Markov chain, and second-order Markov chain, for predicting homoblock frequencies in these polysaccharides. Furthermore, based on the mathematically derived inequality relations, a novel 2D array was constructed, enabling the graphical visualization of homoblock features in polysaccharides. It was demonstrated, as a proof of concept, that the novel 2D array, along with a 1D code generated from it, could serve as an effective feature engineering tool for polymer classification using machine learning algorithms.

Influence of Hydroxycinnamic Acids on the Maillard Reaction of Arabinose and Galactose beyond Carbonyl-Trapping.

Hydroxycinnamic acids, known for their health benefits and widespread presence in plant-based food, undergo complex transformations during high-temperature processing. Recent studies revealed a high browning potential of hydroxycinnamic acids and reactive Maillard reaction intermediates, but the role of phenolic compounds in the early stage of these reactions is not unambiguously understood. Therefore, we investigated the influence of caffeic acid and ferulic acid on the nonenzymatic browning of arabinose, galactose, and/or alanine, focusing on the implications on the formation of relevant early-stage Maillard intermediates and phenol-deriving products. Contrary to previous assumptions, hydroxycinnamic acids were found to promote nonenzymatic browning instead of solely trapping reactive intermediates. This was reflected by an intense browning, which was attributed to the formation of heterogeneous phenol-containing Maillard products. Although, caffeic acid is more reactive than ferulic acid, the formation of reactive furan derivatives and of heterogeneous phenol-containing colorants was promoted in the presence of both hydroxycinnamic acids.

Multilayer Annotation Strategy AnnoSePS: Disentangling the Intricate Structure of Selenium-Containing Polysaccharides Based on Preferential Fragmentation Patterns.

Precision mapping of selenium at structural and position levels poses significant challenges in selenium-containing polysaccharide identification. Due to the absence of reference spectra, database-centric approaches are still limited in the discovery of selenium binding sites and distinction among different isomeric structures. A multilayer annotation strategy, AnnoSePS, is proposed for achieving the identification of seleno-substituent and the unbiased profiling of polysaccharides. Applying Snoop-triggered multiple reaction monitoring (Snoop-MRM) identified multidimensional monosaccharides in selenium-containing polysaccharides. Galactose, galacturonic acid, and glucose were the predominant monosaccharides with a molar ratio of 25.19, 19.45, and 11.72, respectively. Selenium present in seleno-rhamnose was found to substitute the hydroxyl group located at C-1 positions through the formation of a Se-H bond. Ions C6H9O3Se-, C6H7O3Se-, C5H5O3Se-, C4H5O2Se-, C3H5O2Se-, C2H3O2Se-, and CHOSe- were defined as the characteristic fragments of seleno-rhamnose. The agglomerative hierarchical clustering algorithm is applied to group spectra from each run based on the characteristic information. Preferential fragmentation patterns in mass spectrometry are revealed by training a probabilistic model. A list of candidate oligosaccharides is generated by step-by-step browsing through the transition pairs for all reference spectra and applying the transitions (addition, insertion, removal, and substitution) to reference structures. Combining time course analyses revealed the linkage composition of selenium-containing oligosaccharides. Glycosidic linkages were annotated based on a synthesis-driven approach. T-Galactose (16.67 ± 5.23%) and T-Galacturonic acid (11.54 ± 4.66%) were the predominant linkage residues. As the database-independent mapping strategy, AnnoSePS makes it possible to comprehensively interrogate spectral data and dissect the fine structure of selenium-containing polysaccharides.

Fisetin-loaded pluronic-based nanogel: Radiation synthesis for alleviating neurocognitive impairments in a rat model of alzheimer's disease via modulation of the apoptotic cascade.

Alzheimer's disease (AD) is a neurodegenerative disorder marked by cognitive impairment and memory loss. In this study, AD was experimentally induced in rats using aluminum chloride (AlCl3) and D-galactose (D-gal). Fisetin (Fis), a natural compound with antioxidant and anti-inflammatory properties, has potential for neurodegeneration management, but its low bioavailability limits clinical applications. To address this, we synthesized and characterized Pluronic-2-Acrylamido-2-methylpropane sulfonic acid (PLUR-PAMPS) nanogels using gamma radiation and successfully loaded Fis onto them (Fis-PLUR-PAMPS). The optimal formulation exhibited minimal particle size, a highly acceptable polydispersity index, and the highest zeta-potential, enhancing stability and solubilization efficiency. Our goal was to improve Fis's bioavailability and assess its efficacy against AlCl3/D-gal-induced AD. Male albino Wistar rats were pre-treated orally with Fis (40 mg/kg) or Fis-PLUR-PAMPS for seven days, followed by a seven-day intraperitoneal injection of AlCl3 and D-gal. Behavioral assessments, histopathological analysis, and biochemical evaluation of markers related to AD pathology were conducted. Results demonstrated that Fis-PLUR-PAMPS effectively mitigated cognitive impairments and neurodegenerative signs induced by AlCl3/D-gal. These findings suggest that Fis-PLUR-PAMPS nanogels enhance Fis's bioavailability and therapeutic efficacy, offering a promising approach for AD management.

Enzymatically Oxidized Carbohydrates As Dicarbonyl Biobased Cross-Linkers for Polyamines.

Carbonyl cross-linkers are used to modify textiles and form resins, and are produced annually in megatonne volumes. Due to their toxicity toward the environment and human health, however, less harmful biobased alternatives are needed. This study introduces carbonyl groups to lactose and galactose using galactose oxidase from Fusarium graminearum (FgrGalOx) and pyranose dehydrogenase from Agaricus bisporus (AbPDH1) to produce four cross-linkers. Differential scanning calorimetry was used to compare cross-linker reactivity, most notably resulting in a 34 °C decrease in reaction peak temperature (72 °C) for FgrGalOx-oxidized galactose compared to unmodified galactose. Attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy, X-ray photoelectron spectroscopy (XPS), and proton nuclear magnetic resonance (1H NMR) spectroscopy were used to verify imine formation and amine and aldehyde depletion. Cross-linkers were shown to form gels when mixed with polyallylamine, with FgrGalOx-oxidized lactose forming gels more effectively than all other cross-linkers, including glutaraldehyde. Further development of carbohydrate cross-linker technologies could lead to their adoption in various applications, including in adhesives, resins, and textiles.

The study of galactomannans with different molecular weights and their ability to form microparticles suitable for pulmonary delivery.

The influence of locust bean gum (LBG) galactomannans (GMs) molecular weight (Mw) to assemble microparticulate systems was evaluated, and carriers for deep lung delivery were developed. A commercial batch of LBG with a mannose/galactose (M/G) ratio of 2.4 (batch 1) was used to study the influence of different microwave partial acid hydrolysis conditions on carbohydrate composition, glycosidic linkages, and aqueous solutions viscosity. The microwave treatment did not affect the composition, presenting 4-Man (36-42 %), 4,6-Man (27-35 %), and T-Gal (24-25 %) as the main glycosidic linkages. Depolymerization led to a viscosity reduction (≤0.005 Pa·s) with no major impact on polysaccharide debranching. The structural composition of the LBG galactomannans were further elucidated with sequence-specific proteins using carbohydrate microarray technologies. A second batch of LBG (M/G 3.3) was used to study the impact of GMs with different Mw on microparticle assembling, characteristics, and insulin release kinetics. The low-Mw GMs microparticles led to a faster release (20 min) than the higher-Mw (40 min) ones, impacting the release kinetics. All microparticles exhibited a safety profile to cells of the respiratory tract. However, only the higher-Mw GMs allowed the assembly of microparticles with sizes suitable for this type of administration.

Hydrogels based on galactomannan and κ-carrageenan containing immobilized biomolecules for in vivo thermal-burn wound treatment.

Hydrogels based on natural polysaccharides have demonstrated efficacy in epithelial recovery from cutaneous burn wounds. Here, we prepared a double-network hydrogel consisting of galactomannan (from Cassia grandis seeds) and κ-carrageenan (commercially sourced), cross-linked with CaCl2, as a matrix for immobilizing lactoferrin and/or Cramoll, aiming at its applicability as dressings for second-degree burn wounds. The formulations obtained [H - hydrogel, HL - hydrogel + lactoferrin, HC - hydrogel + Cramoll and HLC - hydrogel + lactoferrin + Cramoll] were analyzed rheologically as well as in terms of their stability (pH, color, microbial contamination) for 90 days. The burn was created with an aluminum bar (97 ± 3 °C) in the dorsal region of Wistar rats and subsequently treated with hydrogels (H, HL, HC, HLC) and control saline solution (S). The burn was monitored for 3, 7 and 14 days to evaluate the efficacy of the hydrogels in promoting wound healing. The hydrogels did not reveal significant pH or microbiological changes; there was an increase in brightness and a reduction in opacity for H. The rheological analysis confirmed the gel-like viscoelastic signature of the systems without substantial modification of the basic rheological characteristics, however HLC proved to be more rigid, due to rheological synergy when combining protein biomolecules. Macroscopic analyses confirmed centripetal healing with wound contraction: S < H < HC < HL < HLC. Histopathological analyses showed that hydrogel-treated groups reduced inflammation, tissue necrosis and fibrosis, while promoting re-epithelialization with focal acanthosis, especially in HLC due to a positive synergistic effect, indicating its potential as a promising therapy in the repair of burns.

Galf-Specific Neolectins: Towards Promising Diagnostic Tools.

In the absence of naturally available galactofuranose-specific lectin, we report herein the bioengineering of GalfNeoLect, from the first cloned wild-type galactofuranosidase (Streptomyces sp. strain JHA19), which recognises and binds a single monosaccharide that is only related to nonmammalian species, usually pathogenic microorganisms. We kinetically characterised the GalfNeoLect to confirm attenuation of hydrolytic activity and used competitive inhibition assay, with close structural analogues of Galf, to show that it conserved interaction with its original substrate. We synthetised the bovine serum albumin-based neoglycoprotein (GalfNGP), carrying the multivalent Galf units, as a suitable ligand and high-avidity system for the recognition of GalfNeoLect which we successfully tested directly with the galactomannan spores of Aspergillus brasiliensis (ATCC 16404). Altogether, our results indicate that GalfNeoLect has the necessary versatility and plasticity to be used in both research and diagnostic lectin-based applications.

Bayesian optimization of glycopolymer structures for the interaction with cholera toxin B subunit.

The optimal structure of synthetic glycopolymers for GM1 mimetics was determined through Bayesian optimization. The interactions of glycopolymers carrying galactose and neuraminic acid units in different compositions with cholera toxin B subunit (CTB) were assessed by an enzyme-linked immunosorbent assay (ELISA). Gaussian process regression, using the ELISA results, predicted the composition of glycopolymers that would exhibit stronger interactions with CTB. Following five cycles of optimization, the glycopolymers carrying 60 mol% galactose and 25 mol% neuraminic acid demonstrated an IC50 value of 75 µM for CTB, representing the lowest value among the synthesized glycopolymers.

Acetylated galactopyranosyl N-heterocyclic monocarbene complexes of Silver(I) as novel anti-proliferative agents in a rhabdomyosarcoma cell line.

Herein, four silver(I) complexes bearing acetylated d-galactopyranoside-based N-heterocyclic carbene ligands were synthesized and fully characterized by elemental analysis, NMR, and X-ray photoelectron spectroscopy. All complexes were obtained with an anomeric ß-configuration and as monocarbene species. In this study, we investigated the biological effects of the silver(I) complexes 2a-d on the human rhabdomyosarcoma cell line, RD. Our results show concentration-dependent effects on cell density, growth inhibition, and activation of key signaling pathways such as Akt 1/2, ERK 1/2, and p38-MAPK, indicating their potential as anticancer agents. Notably, at 35.5 µM, the complexes induced mitochondrial network disruption, as observed with 2b and 2c, whereas with 2a, this disruption was accompanied by nuclear content release. These results provide insight into the utility of carbohydrate incorporated NHC complexes of silver(I) as new agents in cancer therapy.

Comprehensive characterization of hemicelluloses obtained from Gleditsia sinensis Lam. pods and the application of moderately degraded hemicelluloses in galactomannan film.

The Gleditsia sinensis Lam. pods (GSP) are consistently discarded as waste after saponin extraction due to a lack of industrial or high-value utilization. Herein, the hemicelluloses were extracted from two varieties of GSP and subjected to comprehensive characterization. The molar mass of DMSO-soluble hemicelluloses (53.3-66.0 kDa) was higher compared to that of alkali-soluble ones (24.9-32.6 kDa). The presence of minimal acetyl substitution (3.85-4.49 %) on xylan was unequivocally confirmed. NMR spectroscopic analysis indicated that the hemicelluloses in GSP predominantly consist of a 1,4-ß-ᴅ-Xyl backbone with arabinose substituents at O-3 and 4-O-methyl-α-ᴅ-GlcA substituents at O-2 of the xylose residues. p-Coumaric acid substitution also occurred on the 1,4-ß-ᴅ-Xyl backbone. Hydrothermal treatment significantly reduced the hemicelluloses' relative molar mass and produced 7-10 % xylo-oligosaccharides. Furthermore, the moderately degraded hemicelluloses exhibited significantly enhanced biological activity. Finally, the incorporation of the moderately degraded hemicelluloses imparted the galactomannan film with exceptional antioxidant properties (81.1 % DPPH scavenging activity), while negligibly affecting its transparency. Our study's findings will contribute to a comprehensive understanding of the structural and biochemical properties of hemicellulose in waste G. sinensis pods, thereby facilitating their enhanced utilization in industrial applications.

Simple and sensitive galactose monitoring based on capillary SERS sensor.

Galactosemia, a severe genetic metabolic disorder, results from the absence of galactose-degrading enzymes, leading to harmful galactose accumulation. In this study, we introduce a novel capillary-based surface-enhanced Raman spectroscopy (SERS) sensor for convenient and sensitive galactose detection. The developed sensor enhances SERS signals by introducing gold nanoparticles (Au NPs) onto the surface of silver nanoshells (Ag NSs) within a capillary, creating Ag NSs with Au NPs as satellites. Utilizing 4-mercaptophenylboronic acid (4-MPBA) as a Raman reporter molecule, the detection method relies on the conversion of 4-MPBA to 4-mercaptophenol (4-MPhOH) driven by hydrogen peroxide (H2O2) generated during galactose oxidation by galactose oxidase (GOx). A new SERS signal was observed, which was generated by H2O2 produced when galactose and GOx reacted. Our strategy yielded a quantitative change in the SERS signal, specifically in the band intensity ratio of 998 to 1076 cm-1 (I998/I1076) as the galactose concentration increased. Our capillary-based SERS biosensor provides a promising platform for early galactosemia diagnosis.