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Cassia mimosoides L. (CM), a member of the family Leguminosae, represents an underexplored component within the Katakakhadiradi Kashayam, an Ayurvedic preparation primarily utilized for the management and treatment of diabetes mellitus. For the first time, the potential of CM to address diabetic complications was scientifically validated using in vitro methods. The significant alpha-glucosidase activity of ethanol-water extract (IC 50 of 1.02 ± 0.12 μg/mL) as against the standard acarbose (IC 50 19.28 ± 1.42 μg/mL) has prompted further column separation to identify the active components. Using bioassay-guided fractionation based on Dipeptidyl Peptidase-IV (DPP-IV) inhibitory activity, four compounds were isolated and identified through NMR and HRMS. All the four compounds, orientin, isoorientin, diosmetin and luteolin exhibited marked inhibition over the DPP-IV enzyme. Among them disometin showed significantly higher activity with an IC 50 of 2.80 ± 0.38 μM. At non-cytotoxic concentrations these compounds effectively stimulated 2-NBDG [2-(7-nitrobenz-2-oxa-1,3-diazol- 4-yl) amino-2-deoxy-d-glucose] uptake in L6 myotubes. Notably, this investigation represents the first report elucidating the glucose uptake-enhancing potential of the flavonoid isoorientin in L6 myoblast cell lines. Moreover, isoorientin also exhibited significant antiglycation activity in assay using bovine serum albumin and methylglyoxal. IC 50 values for isoorientin and positive control aminoguanidine were found to be 51.45 ± 0.03 µg/mL and 45.18 ± 0.03 µg/mL, respectively. In addition, virtual docking studies of the four compounds against α-glucosidase, α-amylase, and DPP-IV demonstrated strong affinity, thus providing further evidence to substantiate the observed activity. Consequently, this study revealed the hypoglycemic efficacy of the plant CM and proposed the exploration of its compounds in ameliorating type 2 diabetes mellitus. [Display omitted] • Cassia mimosoides L. (CM) exhibits promising multi-modal anti-diabetic efficacy. • The proanthocyanidin-rich fraction demonstrated superior inhibition of α-glucosidase and α-amylase. • Isoorientin exhibits enhanced glucose uptake potential for the first time in L6 cell lines. • All the compounds showed significant potential against DPP-IV. [ABSTRACT FROM AUTHOR]

Gingerols are phenolic biomedical compounds found in ginger (Zingiber officinale) whose low aqueous solubility limits their medical application. To improve their solubility and produce novel glucosides, an α -glucosidase (glycoside hydrolase) from Agrobacterium radiobacter DSM 30147 (Ar G) was subcloned, expressed, purified, and then confirmed to have additional α -glycosyltransferase activity. After optimization, the Ar G could glycosylate gingerols into three mono-glucosides based on the length of their acyl side chains. Compound 1 yielded 63.0 %, compound 2 yielded 26.9 %, and compound 3 yielded 4.37 %. The production yield of the gingerol glucosides optimally increased in 50 mM phosphate buffer (pH 6) with 50 % (w/v) maltose and 1000 mM Li+ at 40 °C for an 24-h incubation. The structures of purified compound 1 and compound 2 were determined as 6-gingerol-5- O - α -glucoside (1) and novel 8-gingerol-5- O - α -glucoside (2), respectively, using nucleic magnetic resonance and mass spectral analyses. The aqueous solubility of the gingerol glucosides was greatly improved. Further assays showed that, unusually, 6-gingerol-5- O - α -glucoside had 10-fold higher anti-inflammatory activity (IC 50 value of 15.3 ± 0.5 μM) than 6-gingerol, while the novel 8-gingerol-5- O - α -glucoside retained 42.7 % activity (IC 50 value of 106 ± 4 μM) compared with 8-gingerol. The new α -glucosidase (Ar G) was confirmed to have acidic α -glycosyltransferase activity and could be applied in the production of α -glycosyl derivatives. The 6-gingerol-5- O - α -glucoside can be applied as a clinical drug for anti-inflammatory activity. [Display omitted] [ABSTRACT FROM AUTHOR]

The global incidence of invasive fungal infections (IFIs) has increased over the past few decades, mainly in immunocompromised patients, and is associated with high mortality and morbidity. Aspergillus fumigatus is one of the most common and deadliest IFI pathogens. Major hurdles to treating fungal infections remain the lack of rapid and definitive diagnosis, including the frequent need for invasive procedures to provide microbiological confirmation, and the lack of specificity of structural imaging methods. To develop an Aspergillus-specific positron emission tomography (PET) imaging agent, we focused on fungal-specific sugar metabolism. We radiolabeled cellobiose, a disaccharide known to be metabolized by Aspergillus species, and synthesized 2-deoxy-2-[18F]fluorocellobiose ([18F]FCB) by enzymatic conversion of 2-deoxy-2-[18F]fluoroglucose ([18F]FDG) with a radiochemical yield of 60 to 70%, a radiochemical purity of >98%, and 1.5 hours of synthesis time. Two hours after [18F]FCB injection in A. fumigatus pneumonia as well as A. fumigatus, bacterial, and sterile inflammation myositis mouse models, retained radioactivity was only seen in foci with live A. fumigatus infection. In vitro testing confirmed production of β-glucosidase enzyme by A. fumigatus and not by bacteria, resulting in hydrolysis of [18F]FCB into glucose and [18F]FDG, the latter being retained by the live fungus. The parent molecule was otherwise promptly excreted through the kidneys, resulting in low background radioactivity and high target-to-nontarget ratios at A. fumigatus infectious sites. We conclude that [18F]FCB is a promising and clinically translatable Aspergillus-specific PET tracer. Editor's summary: Diagnosis of invasive Aspergillus infections can be difficult, in part because of lack of specificity of imaging methods. Here, Shah and colleagues rapidly synthesized an Aspergillus-specific 2-deoxy 2-[18F]fluorocellobiose ([18F]FCB) positron emission tomography (PET) tracer. Using mouse models of myositis and pulmonary infection, the authors showed that [18F]FCB injection resulted in retained radioactivity only at sites of live Aspergillus infection, but not at sites of heat-killed spore or bacterial injection, and that radioactivity on PET imaging localized to infiltrates and consolidations seen on coregistered CT scans. Longitudinal [18F]FCB PET/CT imaging of mice with Aspergillus myositis also detected infection progression and subclinical residual infection, highlighting the clinical potential of this Aspergillus-specific PET tracer. —Melissa L. Norton [ABSTRACT FROM AUTHOR]

Starch is a polysaccharide produced exclusively through photosynthesis in plants and algae; however, is utilized as an energy source by most organisms, from microorganisms to higher organisms. In mammals and the germinating seeds of plants, starch is metabolized by simple hydrolysis pathways. Moreover, starch metabolic pathways via unique oligosaccharides have been discovered in some bacteria. Each organism has evolved enzymes responsible for starch metabolism that are diverse in their enzymatic properties. This review, focusing on eukaryotic α-glucosidases and bacterial α-glucoside-hydrolyzing enzymes, summarizes the structural aspects of starch-metabolizing enzymes belonging to glycoside hydrolase families 15, 31, and 77 and their application for oligosaccharide production. [ABSTRACT FROM AUTHOR]

Malvidin-3-O-glucoside (mv-3-O-glc), malvidin-3,5-O-diglucoside (mv-3,5-O-diglc), and malvidin-3-O-(6-O-coumaroyl) -glucoside-5-O-glucoside (mv-3,5-O-diglcCoum) extracted from Yan 73 (Vitis vinifera L.) and spine grape (Vitis davidii Foex) were used to investigate the effect on maltase and sucrase in the gut microbiota and Caco-2 cells. The results showed that three anthocyanins exhibited the inhibition of both maltase and sucrase in Caco-2 cells, and mv-3,5-O-diglc was the most effective one with an IC50 of 258.96 μM for maltase and 189 μM for sucrase, respectively, through a mixed-type inhibitory mechanism. The strong inhibition was also supported by the significant decrease in sucrase mRNA and protein levels in Caco-2 cells after treatment with mv-3,5-O-diglc for 4 h. Moreover, the inhibition effect of anthocyanins was observed only on maltase, which originated from the gut microbiota. The inhibition potential of mv-3-O-glc against maltase was higher than that of the other two anthocyanins in a mixed-type inhibition manner (IC50 = 9.66 ± 0.33 μM). Molecular docking revealed that mv-3-O-glc and mv-3,5-O-diglc formed hydrogen bonds with the aspartic acid (ASP) 443 catalytic residue of maltase, respectively, and then inhibited the enzyme activity by occupying the catalytic center. In particular, mv-3,5-O-diglc could easily fit into the active center of sucrase and form hydrogen bonds with the ASP1394 residue, which could be another reason for the inhibition of sucrase. Thus, anthocyanins, especially the diglucoside anthocyanins derived from grapes, are good potential α-glucosidase inhibitors for regulating postprandial blood glucose levels as dietary supplements. [ABSTRACT FROM AUTHOR]

Functional microbial communities producing β-glucosidases differentially regulate the expression of glucose/non-glucose-tolerant β-glucosidases in response to carbon catabolite repression (CCR), which is a common phenomenon. To better understand their biological roles in a composting environment, four representative β-glucosidase genes were cloned from the metatranscriptome of the compost for expression and characterization. BGLA contains conserved sites Trp168 and Leu173, associated with glucose tolerance. In the presence of 100 mM glucose, BGLA's hydrolysis activity increases by 80%. BGLB also contains Trp168 and Leu173; however, its hydrolysis activity is significantly inhibited by glucose. On the contrary, Trp168 of BGLC was replaced by Phe, yet its hydrolysis activity increased by 20% in the presence of 25 mM glucose. However, BGLD did not exhibit any enzymatic activity. Molecular docking and dynamic simulations revealed the structural basis of glucose tolerance. Metatranscriptomic analysis of the four genes showed that functional microbial communities upregulated or downregulated gene expression in response to different CCR environments to maintain overall carbon metabolic balance. These findings demonstrate that the enzymatic activity characteristics of individual glucose/non-glucose-tolerant β-glucosidases are consistent with their transcriptional regulation under CCR in the complex composting environment. [Display omitted] • Four new GH1 family β-glucosidases cloned from compost. • BGLA activity increased by 80% with added glucose. • Computational biology elucidates β-glucosidase glucose tolerance mechanism. • Glucose tolerance of β-glucosidases correlates with their expression level. [ABSTRACT FROM AUTHOR]

In our continuation of exploring antidiabetic agents from Garcinia species, we found that the methanolic extract of G. macrantha A.C.Sm. exhibited considerable α-glucosidase inhibition of 58.20 ± 0.37% in sucrose substrate and 39.86 ± 2.07% in maltose substrate at 100 μg/mL. Phytochemical investigation on the extract revealed the presence of a new biphenyl, macrabiphenyl A, which was successfully elucidated by means of spectroscopic methods (HRESIMS and 1D and 2D NMR). The α-glucosidase inhibitory evaluation indicated that the new compound was weakly active against the enzyme. [ABSTRACT FROM AUTHOR]

In this study, 2(3),9(10),16(17),23(24)-tetrakis-[(N-methyl-(1-benzylpiperidin-4-yl)oxy)phthalocyaninato]zinc(II) iodide (ZnPc-2) was synthesized and characterized using spectral methods (FT-IR, 1H-NMR, UV-Vis and mass spectroscopy). The interaction of ZnPc-2 with DNA was investigated by using the UV/Vis titrimetric method, thermal denaturation profile, agarose gel electrophoresis and molecular docking studies. Additionally, the antidiabetic activity of ZnPc-2 was revealed spectroscopically by studying α-amylase and α-glucosidase inhibition activities. The spectroscopic results indicated that ZnPc-2 effectively binds to calf thymus-DNA (CT-DNA) with a Kb value of 7.5 × 104 M−1 and interacts with CT-DNA via noncovalent binding mode. Gel electrophoresis results also show that ZnPc-2 binds strongly to DNA molecules and exhibits effective nuclease activity even at low concentrations. Furthermore, docking studies suggest that ZnPc-2 exhibits a stronger binding tendency with DNA than the control compounds ethidium bromide and cisplatin. Consequently, due to its strong DNA binding and nuclease activity, ZnPc-2 may be suitable for antimicrobial and anticancer applications after further toxicological tests. Additionally, antidiabetic studies showed that ZnPc-2 had both α-amylase and α-glucosidase inhibition activity. Moreover, the α-glucosidase inhibitory effect of ZnPc-2 was approximately 3500 times higher than that of the standard inhibitor, acarbose. Considering these results, it can be said that ZnPc-2 is a moderate α-amylase and a highly effective α-glucosidase inhibitor. This suggests that ZnPc-2 may have the potential to be used as a therapeutic agent for the treatment of type 2 diabetes. [ABSTRACT FROM AUTHOR]

One new iridoid namely rupesin F (1) together with four known ones (2–5) were isolated from the roots of Valeriana jatamansi Jones. The structures were established using spectroscopic methods (1D and 2D NMR including HSQC, HMBC, COSY and NOESY) and by comparison with previously published literature data. The isolated compounds 1 and 3 exhibited strong α-glucosidase inhibition activity with IC50 values of 10.13 ± 0.11 and 9.13 ± 0.03 μg/mL, respectively. This study enriched the chemical diversity of metabolites and provides a direction for the development of antidiabetic agents. [ABSTRACT FROM AUTHOR]

Viburnum luzonicum Rolfe is widely used in China as folk medicine. The bioactivity evaluation indicated that the n-BuOH fraction of V. luzonicum leaves (VLLB) could significantly inhibit α‑amylase and α-glucosidase. In order to clarify its active constituents, the phytochemical analysis on VLLB was first performed using HPLC-QTOF-MS/MS, and three new phenolic compounds, viburosides A–C (1–3), along with seven known analogues (4–10) were isolated through preparative HPLC. The undescribed compounds were determined by extensive spectroscopic analyses (1H and 13C NMR, HSQC, HMBC, HRESIMS, and ORD) and enzymatic hydrolysis. In the in vitro enzyme assays, compounds 1–8 showed potent α‑amylase and α-glucosidase inhibitory activities. The enzymatic kinetics and molecular docking of the strongest inhibitors 2 and 3 against the corresponding target enzyme were also performed. [ABSTRACT FROM AUTHOR]

The bovine rumen contains a large consortium of residential microbes that release a variety of digestive enzymes for feed degradation. However, the utilization of these microbial enzymes is still limited because these rumen microorganisms are mostly anaerobes and are thus unculturable. Therefore, we applied a sequence-based metagenomic approach to identify a novel 2,445-bp glycoside hydrolase family 3 β-glucosidase gene known as BrGH3A from the metagenome of bovine ruminal fluid. BrGH3A β-glucosidase is a 92-kDa polypeptide composed of 814 amino acid residues. Unlike most glycoside hydrolases in the same family, BrGH3A exhibited a permuted domain arrangement consisting of an (α/β)6 sandwich domain, a fibronectin type III domain and a (β/α)8 barrel domain. BrGH3A exhibited greater catalytic efficiency toward laminaribiose than cellobiose. We proposed that BrGH3A is an exo-acting β-glucosidase from Spirochaetales bacteria that is possibly involved in the intracellular degradation of β-1,3-/1,4-mixed linkage glucans that are present in grass cell walls. BrGH3A exhibits rich diversity in rumen hydrolytic enzymes and may represent a member of a new clan with a permuted domain topology within the large family. [ABSTRACT FROM AUTHOR]

Introduction: The Bruguiera cylindrica L. is a mangrove plant that is typically found in coastal areas of Asia, including India. It has been known for its medicinal properties, which have been utilized for generations. For example, in Thailand, it has been used to treat wounds and diarrhoea, while in India, it has been effective in addressing diabetes, ulcers, and other health issues. This particular study sought to investigate the potential of B. cylindrica bark extract in reducing the symptoms of diabetes in rats. Methods: In this study, we examined the potential of B. cylindrica bark extract as an inhibitor of α-amylase and α-glucosidase enzymes in vitro. We also evaluated the effects of the extract and Metformin on rats fed high-fat diets and measured their lipid profiles and biochemical parameters. Furthermore, we conducted a network pharmacology analysis to identify proteins and pathways involved in the amelioration of diabetes. Results: Through metabolite profiling, we identified 58 compounds in B. cylindrica hydroalcoholic extract. These compounds include alkaloids, phenolics, flavonoids, and fatty acids. The extract was found to have a dose-dependent inhibition activity against α-amylase and α-glucosidase, with IC50 values similar to acarbose. In rats, oral administration of 200–400 mg/kg of B. cylindrica led to reduced blood glucose levels and normalized serum biochemical parameters. Conclusions: Bruguiera cylindrica bark may reduce blood sugar levels in rats with diabetes. The study found metabolites that interact with protein targets associated with different types of diabetes. Graphical Abstract [ABSTRACT FROM AUTHOR]

Rehmannia glutinosa was extensively used to control blood sugar in diabetes treatment in tradition Chinese medicine. In the present study, three new compounds, including an iridoid rehmannia A (1) and two ionone rehmannias B-C (7-8), together with fourteen known compounds (2-6 and 9-17), were isolated from the roots of R. glutinosa. The structures of these compounds were determined by physicochemical constants and spectral analysis (1D, 2D-NMR and MS). The effect of 1-17 on α-glucosidase activity was tested in vitro. Compounds 9, 10, and 11 (IC50: 5.0, 3.1, and 6.3 mM) showed moderate activity to suppress α-glucosidase relative to acarbose (IC50 = 3.0 mM). The findings provided some new insights to understand the hypoglycemic effect of R. glutinosa and the development towards the α-glucosidase inhibitor drugs. [ABSTRACT FROM AUTHOR]

Psidium guajava is a plant of the Myrtaceae with various pharmacological activity. In this study, the water extract and the isolated compounds from guava leaves were evaluated for in vitro α-glucosidase inhibition using spectrophotometric method. Ellagic acid, quercetin, quercetin-3-O-glucuronide, avicularin, isoquercitrin, and quercetin-3-galactoside showed α-glucosidase inhibitory activity, and their IC50 value were 25.0, 41.0, 53.5, 46.9, 60.0 and 72.1 μg/mL, respectively compared with the positive control acarbose (IC50 49.2 μg/mL). This study could provide a theoretical basis for the application of Psidium guajava in the treatment of hyperglycemia. [ABSTRACT FROM AUTHOR]

Type 2 diabetes mellitus (T2DM) has become a serious public health problem both in our country and worldwide, being the most prevalent type of diabetes. The combined use of drugs in the treatment of T2DM leads to serious side effects, including gastrointestinal problems, liver toxicity, hypoglycemia, and treatment costs. Hence, there has been a growing emphasis on drugs that demonstrate dual interactions. Several studies have suggested that dual-target agents for peroxisome proliferator-activated receptor-γ (PPAR-γ) and alpha-glucosidase (α-glucosidase) could be a potent approach for treating patients with diabetes. We aim to develop new antidiabetic agents that target PPAR-γ and α-glucosidase enzymes using molecular modeling techniques. These compounds show dual interactions, are more effective, and have fewer side effects. The molecular docking method was employed to investigate the enzyme-ligand interaction mechanisms of 159 newly designed compounds with target enzymes. Additionally, we evaluated the ADME properties and pharmacokinetic suitability of these compounds based on Lipinski and Veber's rules. Compound 70, which exhibited favorable ADME properties, demonstrated more effective binding energy with both PPAR-γ and α-glucosidase enzymes (-12,16 kcal/mol, -10.07 kcal/mol) compared to the reference compounds of Acetohexamide (-9.31 kcal/mol, -7.48 kcal/mol) and Glibenclamide (-11.12 kcal/mol, -8.66 kcal/mol). Further, analyses of MM/PBSA binding free energy and molecular dynamics (MD) simulations were conducted for target enzymes with compound 70, which exhibited the most favorable binding affinities with both enzymes. Based on this information, our study aims to contribute to the development of new dual-target antidiabetic agents with improved efficacy, reduced side effects, and enhanced reliability for diabetes treatment. [ABSTRACT FROM AUTHOR]

Trichoderma erinaceum is a filamentous fungus that was isolated from decaying sugarcane straw at a Brazilian ethanol biorefinery. This fungus shows potential as a source of plant cell wall-degrading enzymes (PCWDEs). In this study, we conducted a comprehensive multiomics investigation of T. erinaceum to gain insights into its enzymatic capabilities and genetic makeup. Firstly, we performed genome sequencing and assembly, which resulted in the identification of 10,942 genes in the T. erinaceum genome. We then conducted transcriptomics and secretome analyses to map the gene expression patterns and identify the enzymes produced by T. erinaceum in the presence of different substrates such as glucose, microcrystalline cellulose, pretreated sugarcane straw, and pretreated energy cane bagasse. Our analyses revealed that T. erinaceum highly expresses genes directly related to lignocellulose degradation when grown on pretreated energy cane and sugarcane substrates. Furthermore, our secretome analysis identified 35 carbohydrate-active enzymes, primarily PCWDEs. To further explore the enzymatic capabilities of T. erinaceum, we selected a β-glucosidase from the secretome data for recombinant production in a fungal strain. The recombinant enzyme demonstrated superior performance in degrading cellobiose and laminaribiose compared to a well-known enzyme derived from Trichoderma reesei. Overall, this comprehensive study provides valuable insights into both the genetic patterns of T. erinaceum and its potential for lignocellulose degradation and enzyme production. The obtained genomic data can serve as an important resource for future genetic engineering efforts aimed at optimizing enzyme production from this fungus. [ABSTRACT FROM AUTHOR]

The Ruminiclostridium thermocellum cellulosome system is one of the most efficient cellulase systems in nature. The main hydrolysis product of cellulosic saccharification by R. thermocellum was cellobiose, which served as a strong feedback inhibitor to the cellulosome. In this study, R. thermocellum M3, which directly hydrolyzes cellulosic biomass into monosaccharides, was used to degrade cellobiose to investigate the kinetic characteristics of cellobiose degradation. Moreover, the transcriptomic characteristics of strain M3 under cellobiose and Avicel culture conditions were compared. The results indicated that strain M3 was able to grow and hydrolyze cellobiose under cellobiose concentrations higher than 20 g/L. The best growth performance and glucose production were obtained under 12 g/L cellobiose. Compared with that of Avicel, cellobiose had greater β-glucosidase activity under cellobiose culture, with a maximum glucose yield of 767.97±19.13 mg/g cellobiose. The transcription results indicated that R. thermocellum M3 resisted the feedback inhibition of cellobiose by regulating genes encoding β-glucosidase, cellobiose phosphorylase and ABC transporters. The high glucose yield of strain M3 was closely related to the expression of the bglX gene. In addition, the gene related to the chemotactic pathway of strain M3 also increased its perception and adaptation to cellobiose. [Display omitted] • The cellulosome of R. thermocellum M3 remains active under cellobiose more than 20 g/L. • R. thermocellum hydrolyzed cellobiose related with BGL and non-BGL genes. • BglX was potential responsible for the cellobiose hydrolysis by R. thermocellum. • The findings provide solutions for the biorefined of cellulosic energy and fuels. [ABSTRACT FROM AUTHOR]

Progesterone has been shown to stimulate glycogen catabolism in uterine epithelial cells. Acid α‐glucosidase (GAA) is an enzyme that breaks down glycogen within lysosomes. We hypothesized that progesterone may stimulate glycogenolysis in the uterine epithelium via GAA. We found that GAA was more highly expressed in the stroma on Day 1 than on Day 11. However, GAA did not appear to differ in the epithelium on Days 1 and 11. Progesterone (0‐10 μM) had no effect on the levels of the full‐length inactive protein (110 kDa) or the cleaved (active) peptides present inside the lysosome (70 and 76 kDa) in immortalized bovine uterine epithelial (BUTE) cells. Furthermore, the activity of GAA did not differ between the BUTE cells treated with 10 μM progesterone or control. Overall, we confirmed that GAA is present in the cow endometrium and BUTE cells. However, progesterone did not affect protein levels or enzyme activity. [ABSTRACT FROM AUTHOR]

The study aims to analyse the qualitative characteristics of safflower oil, including its fatty acid profile, and determine the acid and peroxide numbers. The study also investigated the levels of heavy metals in the oil and assessed its potential as an inhibitor of α-amylase and α-glucosidase enzymatic activity. In this study, standardised methods were used to analyse the fatty acid composition of safflower oil using a standard mixture of 37 fatty acid methyl esters (Supelco™ 37 Component FAME Mix), which ensured high accuracy in determining the fatty acid composition. The physical properties of safflower oil, such as density (0.94) and specific gravity (0.917), depend on the fatty acid composition and temperature. The refractive index, determined at 1.469, was used to estimate changes in the oil's unsaturation and viscosity (45.6 cP). Colour parameters (lightness 45.24, red-green scale -2.87, yellowblue scale 21.04) confirm the authenticity of the oil. The results of gas chromatographic analysis confirmed that the composition of the samples did not meet the requirements of the standard. The study also included the evaluation of the inhibitory activity of safflower oil against the enzymes α-amylase and α-glucosidase, where one of the samples showed significant inhibition of α-amylase activity and the other α-glucosidase activity, indicating their potential use as natural inhibitors. The safety control of heavy metal content showed that the levels of lead, cadmium and other toxic elements in the oil are well below the permissible limits, confirming its safety for consumption. These data indicate that, although some safflower oil samples do not meet the standards for fatty acid composition, its potential as a natural enzyme inhibitor and safety in terms of heavy metal content opens up prospects for further research and possible optimisation of production processes. [ABSTRACT FROM AUTHOR]

Epigallocatechin gallate (EGCG) has an effect in reducing sugar levels in the blood by inhibiting α-glucosidase enzyme, which is connected explicitly by hydrogen bonds and modifies the secondary structure and micro-environment of the enzyme reversibly and noncompetitive. This study looks at the activity and interaction of EGCG as α-glucosidase inhibitors in the form of binding affinity and compound bonding profiles with receptors, including toxicity predictions and drug-likeness results. The research was performed in silico with molecular docking on Autodock Vina that integrated through PyRx, then viewed the compound's binding profile with receptor using Discovery Studio 2021 Client, toxicity prediction using ProTox-II and determination of drug-likeness using SwissADME based on Lipinski's rule of five guidelines. The control drugs used were acarbose and miglitol. The molecular docking results obtained that the binding affinity of EGCG is -8.4 kcal/mol while acarbose and miglitol are -13.8 kcal/mol and -5.3 kcal/mol respectively. There are amino acid residues similar to the drug control with various interactions like electrostatic, hydrophobic, and hydrogen bonds; then it has an inactive target for each toxicity parameter and has a molecular weight of 458.37 g/mol; Log P value of 1.01; H-bond donor of 8; and H-bond acceptor of 11 in the determination of drug-likeness. Based on these results, EGCG has effectiveness as α-glucosidase inhibitors predicted to be non-toxic; however, there are violations in determining drug-likeness. [ABSTRACT FROM AUTHOR]

Breonadia salicina (Vahl) Hepper & J.R.I. Wood is widely distributed throughout Africa. It is used ethnobotanically to treat various diseases. However, the metabolic profile of the Breonadia species is not well characterized and the metabolites that are responsible for the bioactivity of this plant remain unknown. Therefore, there is a need to determine the phytochemical and bioactivity profile to identify metabolites that contribute to the antidiabetic, anti-inflammatory and antiproliferation activity, including the genotoxicity and cytotoxic effects, of Breonadia salicina. The study is aimed at exploring the metabolomic profile antidiabetic, anti-inflammatory and antiproliferation activity, as well as the genotoxicity and cytotoxicity effects, of constituents of B. salicina. The compounds in the B. salicina extract were analyzed by ultra-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS), and the resultant data were further analyzed using a molecular networking approach. The crude stem bark and root extracts showed the highest antidiabetic activity against α-amylase at the lowest test concentration of 62.5 µg/mL, with 74.53 ± 0.74% and 79.1 ± 1.5% inhibition, respectively. However, the crude stem bark and root extracts showed the highest antidiabetic activity against α-glucosidase at the lowest test concentration of 31.3 µg/mL, with 98.20 ± 0.15% and 97.98 ± 0.22% inhibition, respectively. The crude methanol leaf extract showed a decrease in the nitrite concentration at the highest concentration of 200 µg/mL, with cell viability of 90.34 ± 2.21%, thus showing anti-inflammatory activity. No samples showed significant cytotoxic effects at a concentration of 10 µg/mL against HeLa cells. Furthermore, a molecular network of Breonadia species using UPLC-QTOF-MS with negative mode electrospray ionization showed the presence of organic oxygen compounds, lipids, benzenoids, phenylpropanoids and polyketides. These compound classes were differentially distributed in the three different plant parts, indicating the chemical differences between the stem bark, root and leaf extracts of B. salicina. Therefore, the identified compounds may contribute to the antidiabetic and anti-inflammatory activity of Breonadia salicina. The stem bark, root and leaf extracts of B. salicina yielded thirteen compounds identified for the first time in this plant, offering a promising avenue for the discovery of new lead drugs for the treatment of diabetes and inflammation. The use of molecular networking produced a detailed phytochemical overview of this Breonadia species. The results reported in this study show the importance of searching for bioactive compounds from Breonadia salicina and provide new insights into the phytochemical characterization and bioactivity of different plant parts of Breonadia salicina. [ABSTRACT FROM AUTHOR]

Galacto-oligosaccharides (GOS) are prebiotic compounds that are mainly used in infant formula to mimic bifidogenic effects of mother's milk. They are synthesized by β-galactosidase enzymes in a trans-glycosylation reaction with lactose. Many β-galactosidase enzymes from different sources have been studied, resulting in varying GOS product compositions and yields. The in vivo role of these enzymes is in lactose hydrolysis. Therefore, the best GOS yields were achieved at high lactose concentrations up to 60%wt, which require a relatively high temperature to dissolve. Some thermostable β-glucosidase enzymes from thermophilic bacteria are also capable of using lactose or para nitrophenyl-galactose as a substrate. Here, we describe the use of the β-glucosidase BglA from Thermotoga maritima for synthesis of oligosaccharides derived from lactose and cellobiose and their detailed structural characterization. Also, the BglA enzyme kinetics and yields were determined, showing highest productivity at higher lactose and cellobiose concentrations. The BglA trans-glycosylation/hydrolysis ratio was higher with 57%wt lactose than with a nearly saturated cellobiose (20%wt) solution. The yield of GOS was very high, reaching 72.1%wt GOS from lactose. Structural elucidation of the products showed mainly β(1 → 3) and β(1 → 6) elongating activity, but also some β(1 → 4) elongation was observed. The β-glucosidase BglA from T. maritima was shown to be a very versatile enzyme, producing high yields of oligosaccharides, particularly GOS from lactose. Key points: • β-Glucosidase of Thermotoga maritima synthesizes GOS from lactose at very high yield. • Thermotoga maritima β-glucosidase has high activity and high thermostability. • Thermotoga maritima β-glucosidase GOS contains mainly (β1-3) and (β1-6) linkages. [ABSTRACT FROM AUTHOR]

Beta-glucosidases catalyze the hydrolysis of the glycosidic bonds of cellobiose, producing glucose, which is a rate-limiting step in cellulose biomass degradation. In industrial processes, β-glucosidases that are tolerant to glucose and stable under harsh industrial reaction conditions are required for efficient cellulose hydrolysis. In this study, we report the molecular cloning, Escherichia coli expression, and functional characterization of a β-glucosidase from the gene, CelGH3_f17, identified from metagenomics libraries of an Ethiopian soda lake. The CelGH3_f17 gene sequence contains a glycoside hydrolase family 3 catalytic domain (GH3). The heterologous expressed and purified enzyme exhibited optimal activity at 50 °C and pH 8.5. In addition, supplementation of 1 M salt and 300 mM glucose enhanced the β-glucosidase activity. Most of the metal ions and organic solvents tested did not affect the β-glucosidase activity. However, Cu2+ and Mn2+ ions, Mercaptoethanol and Triton X-100 reduce the activity of the enzyme. The studied β-glucosidase enzyme has multiple industrially desirable properties including thermostability, and alkaline, salt, and glucose tolerance. [ABSTRACT FROM AUTHOR]

Rosa damascena Mill. is widely known for its essential oil. The root part of this plant is not explored. Fresh, shade-dried roots of R. damascena were used for hydrodistillation and a 0.02 % (v/w) yield of volatile organic compounds (VOCs) was obtained. The chemical profile of VOCs was determined by gas chromatography-mass spectrometry (GC–MS) and further quantified by a gas chromatography-flame ionization detector (GC-FID). Screening for antidiabetic potential has been conducted by determining inhibitory effect of α-amylase and α-glucosidase with IC 50 values of 0.95±0.20 mg/mL and 0.92±0.11 mg/mL, respectively. In-vitro studies for the determination of antimicrobial potential against, three Gram-positive, three Gram-negative pathogenic bacterial strains, and two fungal pathogenic strains were also conducted, which showed promising results. Among all the tested pathogenic bacterial strains, Micrococcus luteus MTCC 2470 was found highly sensitive against VOCs with the inhibition zone diameter of 41 ± 1 mm and the minimum inhibitory concentration (MIC) of 0.39 mg/mL. Further, out of two tested pathogenic fungal strains, VOCs showed the highest inhibition potential against Fusarium graminearum MTCC 1893 with the inhibition zone diameter of 39.33±1.53 mm. This study revealed that hexadecane (12.06 %), octadecane (10.73 %), n-hexadecanoic acid, (8.32 %) tetradecane (7.93 %), tetradec‑1-ene (7.53 %), icosane (7.49 %), and hexadec‑1-ene (7.47 %), were the most abundant volatile components. In silico study has been conducted for antidiabetic and antimicrobial. However, after 10–12 years of planting, uprooting is done. After uprooting, the root part is usually burned or disposed. In this study, we demonstrated a possible utilization of root part by extracting VOCs and evaluated their biological significance. [Display omitted] • Rosa damascena Mill. is well known species and it is globally cultivated. • After uprooting of plant, root becomes the waste, usually burned, or discarded. • Around 2,49,750 tonnes of uprooting is done in every decade. • Qualitative and quantitative chemical profiling of VOCs present in root part. • Antidiabetic and antimicrobial potential studied through In-vitro and In-silico. [ABSTRACT FROM AUTHOR]

In this study, the polysaccharide (RHCP) extracted from Houttuynia cordata rhizome was acetylated through the acetic anhydride method. The physicochemical properties of RHCP and its acetylated derivatives (Ac‐RHCP) were determined by infrared spectra, scanning electron microscopy, and Congo red test. Meanwhile, the α‐glucosidase inhibition mechanism of RHCP and Ac‐RHCP was analyzed by inhibition kinetics, and circular dichroism and fluorescence spectroscopy. Ac‐RHCP resulted in a more porous surface structure and 1.83‐fold higher solubility compared with RHCP. At a concentration of 6 mg/mL, the α‐glucosidase inhibition rate of Ac‐RHCP was 75.40%, while that of RHCP was 44.68%. RHCP and Ac‐RHCP inhibited α‐glucosidase in a mixed‐type manner, reduced the endogenous fluorescence of α‐glucosidase, affected the microenvironment of amino acid residues, and changed the conformation of α‐glucosidase. The study indicates that Ac‐RHCP exhibits a certain level of α‐glucosidase inhibition, demonstrating its potential as a functional food for glycemic control. [ABSTRACT FROM AUTHOR]

β-Glucosidases are widely applied to biofuel production, food industry, and improvement in flavors. The novel β-glucosidase gene debgl was identified from the Arctic bacterium Devosia sp. Arc12 and expressed in Escherichia coli BL21 (DE3). The putative gene, belonging to the glycoside hydrolase family 1, consists of 1311 nucleotides and encodes 437 amino acid residues. The optimal temperature and pH of the recombinant DeBgl were 55 °C and 7, respectively. The recombinant DeBgl displayed salt and ethanol tolerance, and the V max , K m , and k cat for the substrate cellobiose were 754 μmol min−1 mg−1, 12.8 mM, and 661.4 s−1, respectively. The concentration of glucose was reached 51.6 g L−1 after incubation at 40 °C for 8 h in 100 g L−1 cellobiose solution. It suggests that the recombinant DeBgl is a good candidate enzyme to degrade complex celluloses. The signal peptide (SP) citH has been proven to be the most suitable for the secretory expression of β-glucosidase by screening the signal peptide library in Bacillus subtilis. The secretory activity of the SP citH fused DeBgl in the vector pMA5 was 9-fold higher than that of the DeBgl without signal peptide. This secretory expression system of β-glucosidase may benefit its industrial application. [Display omitted] • A novel β-glucosidase was isolated from the genus Devosia. • The recombinant DeBgl was highly tolerant to many types of chemicals. • The recombinant DeBgl is a good candidate to degrade complex celluloses. • DeBgl was expressed in Bacillus subtilis WB600 and could be secreted. • Signal peptide citH was the best in DeBgl in WB600. [ABSTRACT FROM AUTHOR]

An extract from the rhizomes of Cassumunar ginger (Zingiber purpureum Roscoe). was found to have significant α-glucosidase inhibitory activity with an IC50 value of 6.3 µg/mL. Two new phenylbutenoids, cassudimin A (1) and cassumunol N (2), and seven known compounds (3–9) were isolated. Their structures and relative configurations of two new compounds were elucidated based on spectra interpretation. Compounds 1–3, 6–9 showed more potent α-glucosidase inhibitory activity than a positive control, acarbose (IC50 = 168.0 µM). Dehydrozingerone (6) exhibited the most potent α-glucosidase inhibition with an IC50 value of 8.3 μM. Compounds 7 and 9 were found in Z. purpureum rhizomes for the first time. [ABSTRACT FROM AUTHOR]

This study aimed to establish a novel method for catalysing salidroside synthesis by immobilized β-D-glucosidase with deep eutectic solvents (DESs) as a new green non-aqueous medium. To this end, the catalytic properties of β-D-glucosidase (i.e. activity, stability, and enzymatic properties) were analysed within sixteen different DESs systems. Although most DESs significantly improved enzyme stability, they also reduced enzymatic activity. This was verified at the molecular level by employing fluorescence spectroscopy to evaluate conformational changes in β-D-glucosidase molecules. Subsequently, chitosan microspheres were used to immobilize β-D-glucosidase; the structures of the microspheres and immobilized enzyme were observed by scanning electron microscopy (SEM). Through single factor experiments and application of a response surface design, the optimal conditions for production of salidroside by the reaction of tyrosol with immobilized β-D-glucosidase were established. These conditions comprised 80 vol% ChCl/G (1:2), 50 °C reaction temperature, 5.8 pH, 100 h reaction time, 45 U/mL enzyme dosage, and a tyrosol/D-glucose molar ratio of 10. Under these conditions, the average conversion rate of the tyrosol substrate was 31.6%. Moreover, a salidroside crude product was obtained with a purity >70% (g/g) and a substrate conversion rate of immobilized β-D-glucosidase >50% of its initial value after five repeated uses. Collectively, this study establishes a novel green reaction system for the efficient preparation of salidroside and other glycoside compounds, while also providing a theoretical foundation for the analysis of β-D-glucosidase catalytic reactions in DESs systems. [ABSTRACT FROM AUTHOR]

Type 2 diabetes mellitus is a metabolic dreadful disease caused by an uncontrolled glucose level in the bloodstream, particularly high after a meal. Inhibitors of glucosidases, involved in the digestion of carbohydrates, can regulate this post‐prandial increase in glucose concentration. The traditional drugs act as competitive inhibitors of both pancreatic α‐amylase and α‐glucosidases and this unselective inhibition is behind severe gastrointestinal side effects related to the concomitant inhibition of α‐amylase. We described herein some perglycosylated cyclodextrins as efficient and selective inhibitors of α‐glucosidase with low micromolar IC50 (3.64‐7.98 μM) compared to the acarbose (IC50 212 μM), clinically used for patients suffering from type 2 diabetes. On the other hand, they do not inhibit α‐amylase (IC50>500 μM). Structure/activity relationship rationalization suggests multiple interactions between the described inhibitors and α‐glucosidase, which support the existence of both active site and allosteric interactions. [ABSTRACT FROM AUTHOR]

The current research focused on enhancing the extraction and purification processes of triterpenoids found in persimmon leaves, as well as assessing their biological effects. The extraction yield of triterpenoids was 8.26%. Activated carbon was used to purify the extracts, resulting in an increase in the content of triterpenoids in the final product from 24.43% to 79.66%. The recovery yield of triterpenoids from the ethanol extract was 73.57%. Ethanol was recovered and reused as a solvent. The total triterpenoids exhibited cytotoxic activities against several cancer cell lines, with IC 50 values of approximately 32.49–55.02 μg/mL, as well as anti-inflammatory activities (IC 50 = 29.55 ± 1.67 μg/mL). The total extract and total triterpenoids demonstrated significant inhibition against α-glucosidase, having IC 50 values of 0.18 ± 0.05 μg/mL and 2.12 ± 0.15 μg/mL, respectively. Additionally, for antioxidant activity, the IC 50 values of the total extract and total triterpenoids were 12.53 ± 0.20 and 13.92 ± 0.18 μg/mL, respectively. The triterpenoids showed higher inhibition of acetylcholinesterase compared with the total extract (IC 50 : 40.07 ± 2.54 and 112.22 ± 9.68 μg/mL, respectively). However, they did not show inhibitory activity against bacteria or fungi. In conclusion, information about the extraction, purification and biological activities of total triterpenoids from persimmon leaves was fully provided for the first time. [Display omitted] • Triterpenoids of persimmon leaves were extracted, purified, and tested of bioactivities. • Extraction yield of total triterpenoids was determined to be 8.26%. • Total triterpenoids were purified from 24.43% to 79.66%. • Ethanol was recovered and reused at least 3 times to extract triterpenoids. • Bioactivities of triterpenoids show potentials in drug development. [ABSTRACT FROM AUTHOR]

Lactiplantibacillus plantarum SN13T is a probiotic plant-derived lactic acid bacterium that can grow in various medicinal plant extracts. In this study, we fermented an aqueous extract of gardenia fructus, the fruit of a medicinal plant, with SN13T, such that the bioactivity of the extract was potentiated after fermentation to suppress the release of inflammatory mediators, such as nitric oxide (NO), reactive oxygen species (ROS), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α), as well as downregulate inflammatory genes in lipopolysaccharides (LPS)-stimulated RAW 264.7 cells. This increased antioxidant and anti-inflammatory activity was mediated through bioconversion of the iridoid glycoside geniposide to its aglycone genipin via the supposed hydrolytic action of β-glucosidases harbored by SN13T. In the complete genome of SN13T, ten putative genes encoding β-glucosidases of glycosyl hydrolase (GH) family 1 organized among eight gene operons were identified. Transcriptional profiling revealed that two 6-phospho-β-glucosidase genes, pbg9 and SN13T_1925, located adjacently in the gene operon SN13T_1923, were transcribed significantly more than the remaining genes during fermentation of the gardenia extract. This suggests the role of these β-glucosidases in bioconversion of geniposide to genipin and the subsequent enhanced bioactivity of the gardenia fructus extract after fermentation with SN13T. [ABSTRACT FROM AUTHOR]

Nanozymes are nanomaterials with intrinsic enzyme-mimic activity, but their large-scale application is generally limited by their low catalytic activity. Herein, we demonstrated that highly exposed Cu active sites on twodimensional (2D) nitrogen-doped carbon (Cux/NC) can serve as efficient peroxidase-like (POD) catalysts with high atomic utilization. Specially, the uniformly distributed Cu active sites could react with H2 O2 to produce singlet oxygen (¹O2) under acidic conditions, which can efficiently oxidizes colorless 3,3 ', 5,5 ' -tetramethylbenzidine (TMB) to blue oxidized TMB (oxTMB). Among various Cux/NC nanozymes studied, the Cu0.14 /NC exhibited smaller maximum catalytic velocities (Vmax) and Menten constant (Km) for TMB and H2 O2. Benefiting from the highly active peroxidase-like activity, the Cu0.14 /NC nanozyme could be successfully applied for the hydroquinone (HQ) and ascorbic acid (AA) detection applications through the inhibitory effect of HQ and AA. More interestingly, α-glucosidase (α-Glu) detection sensing platform could be constructed based on HQ as a signal transmitter, with the detection range ranging from 0 to 12 U/L and the minimum detection limit being 0.68 U/L. This work provides not only an idea for the rational design of highly exposed Cu active sites but also fabricate an effective detection sensing platform for HQ, AA, and α-Glu detection. [ABSTRACT FROM AUTHOR]

Background. Fermented soy foods exhibit the capability to inhibit the α-glucosidase enzyme. The bacteria isolated from these fermented soy foods may contribute to the production of a higher quantity of α-glucosidase inhibitor (α-GI) in optimized condition. Aim. The present study aims to isolate α-glucosidase inhibitor producing bacterial strain from peruyaan, a traditional fermented soy food of Arunachal Pradesh, optimize the production of α-GI for maximum yield, and assess the compounds. Result. Bacillus subtilis PM NEIST_4 (identified by 16S rRNA gene sequencing) was isolated as a potential strain that exhibit maximum α-glucosidase inhibition. This strain showed maximum α-GI productivity in Mueller Hinton Broth (MHB). The cultivation parameters for fermentation of MHB were optimized in order to maximize the yield of α-GIs. The α-GI productivity of 1616.613 ± 84.54 U/mL was observed before optimization, which increased by almost three times after optimization (4808.324 ± 13 U/mL). In vitro assays against α-glucosidase enzyme revealed a significant IC50 value of 596.532 ± 44.80 μg/mL after the optimized study compared to that before optimization (IC50 1705.617 ± 43.95 ug/mL). Besides, α-glucosidase enzyme inhibitory property of fermented MHB (FMHB) was found to be unaffected at varied range of pH and temperature. Oral gavaging of FMHB in sucrose-loaded Wistar rat exhibited maximum reduction in postprandial blood glucose at 400 mg/kg body weight. Furthermore, FMHB was fractionated using solvents of increasing polarity including n-hexane, ethyl acetate, and n-butanol. Ethyl acetate fraction that exhibited the strongest inhibitory action against the α-glucosidase enzyme (IC50 value of 251.55 ± 19.65 μg/mL), showed the presence of pyrrolopyrazine derivatives through GC-MS and LC-MS analyses. Conclusion. The present study demonstrated the isolation of a potent α-GI producing bacterial strain from traditional fermented soy food, and the optimization of the bacterial fermentation process was achieved to maximize the yield of α-GI. These findings underscore promising industrial potential of B. subtilis PM NEIST_4, advocating the use of indigenous microbial resources for the production of α-GI. [ABSTRACT FROM AUTHOR]

β-glucosidase is an essential enzyme for the enzymatic hydrolysis of lignocellulosic biomass, as it catalyzes the final stage of cellulose breakdown, releasing glucose. This paper aims to produce β-glucosidase from Saccharomyces cerevisiae and evaluate the enzymatic degradation of delignified sugarcane bagasse. S. cerevisiae was grown in yeast peptone dextrose medium. Partial purification of the enzyme was achieved through precipitating proteins with ethanol, and the optimal activity was measured by optimizing pH and temperature. The effects of ions, glucose tolerance, and heat treatment were evaluated. Delignified sugarcane bagasse was hydrolyzed by the enzyme. β-glucosidase showed a specific activity of 14.0712 ± 0.0207 U mg−1. Partial purification showed 1.22-fold purification. The optimum pH and temperature were 6.24 and 54 °C, respectively. β-glucosidase showed tolerance to glucose, with a relative activity of 71.27 ± 0.16%. Thermostability showed a relative activity of 58.84 ± 0.91% at 90 °C. The hydrolysis of delignified sugarcane bagasse showed a conversion rate of 87.97 ± 0.10% in the presence of Zn2+, an ion that promoted the highest increase in enzymatic activity. S. cerevisiae produced an extracellular β-glucosidase with good stability at pH and temperatures conventionally applied in the hydrolysis of lignocellulosic biomass, showing viability for industrial application. [ABSTRACT FROM AUTHOR]

Fructose is the most preferred sugar to provide benefits for sweetening and health. As many industrial enzymes are used to produce High Fructose Syrup (HFS), it is vital to explore alternative enzymes for fructose production. Oligo-α-1,6-glucosidase (O-1-6-glucosidase) hydrolyzes non-reducing ends of isomaltooligosaccharides, panose, palatinose, and an a-limit dextrin by breaking α-1,6-glucoside bonds, although it generally has no activity on α-1,4-glucoside bonds of maltooligosaccharides. In this study, sucrose-hydrolyzing activity of O-1-6-glucosidase of thermophilic A. gonensis was evaluated. For this purpose, O-1-6-glucosidase gene region of A. gonensis was cloned in the pET28(a)+ expression vector, the expression product was purified, modeled, and biochemically characterized. The optimal activity of the enzyme was to be at pH 7.0 and 60 °C. The enzyme activity was halved at the end of the 276th h at 60 °C. The enzyme maintained its activity even after 300 h at pH 6.0-10.0. The values of Km, Vmax, kcat, and kcat/Km were determined as 44.69 ± 1.27 mM, 6.28 ± 0.05 µmoL/min/mg protein, 6.70 1/s and 0.15 1/mMs−1, respectively. While Zn2+, Cu2+, Pb2+, Ag2+, Fe3+, Hg2+, and Al2+ metal ions inhibited O-1-6-glucosidase, Mn2+, Fe2+, and Mg2+ ions activated the enzyme. Consequently, A. gonensis O-1-6-glucosidase (rAgoSuc2) has interesting properties, especially for HFS production. [ABSTRACT FROM AUTHOR]

Introduction: Computational simulation study was carried out on bioactive compounds of Ganoderma lucidum (G. lucidum). Methods: Molecular docking and molecular dynamics (MD) simulations were performed. The input files for protein and ligands were retrieved from Protein Data Bank (PDB) and PubChem database. Human maltase-glucoamylase (PDB ID: 3L4Y) is the protein (a-glucosidase enzyme). The ligands are thirteen compounds derived from G. lucidum together with acarbose and miglitol as controls. Results: Docking result showed the lowest binding energy is from Ganomycin B (-7.8 kcal/mol) compared to acarbose and miglitol (-5.0 kcal/mol and -4.4 kcal/mol) respectively. MD simulation showed interaction of 3L4Y-Ganomycin B achieved stable interaction and conformation as follows: root mean square deviation (RMSD) is ± 2.7 Å, average distance of ±1.8 Å and constant hydrogen bonds around 1 - 3. Conclusion: Ganomycin B was found to have good binding affinity, embarking its potential as a potent α-glucosidase inhibitor. [ABSTRACT FROM AUTHOR]

As a crucial enzyme for cellulose degradation, β-glucosidase finds extensive applications in food, feed, and bioethanol production; however, its potential is often limited by inadequate thermal stability and glucose tolerance. In this study, a functional gene (lq-bg5) for a GH1 family β-glucosidase was obtained from the metagenomic DNA of a hot spring sediment sample and heterologously expressed in E. coli and the recombinant enzyme was purified and characterized. The optimal temperature and pH of LQ-BG5 were 55 °C and 4.6, respectively. The relative residual activity of LQ-BG5 exceeded 90% at 55 °C for 9 h and 60 °C for 6 h and remained above 100% after incubation at pH 5.0–10.0 for 12 h. More importantly, LQ-BG5 demonstrated exceptional glucose tolerance with more than 40% activity remaining even at high glucose concentrations of 3000 mM. Thus, LQ-BG5 represents a thermophilic β-glucosidase exhibiting excellent thermal stability and remarkable glucose tolerance, making it highly promising for lignocellulose development and utilization. [ABSTRACT FROM AUTHOR]

Objectives: Silver nanoparticles (AgNPs) are gaining increasing attention in biomedical applications due to their unique properties. Green synthesis methods are environmentally friendly and have demonstrated potential for AgNP production. This study explores the green synthesis of AgNPs using the methanolic extract of Euphorbia milii, a plant known for its medicinal properties. The primary objectives of this research were to synthesize AgNPs using E. milii extract, characterize the nanoparticles (NPs) using various techniques, and evaluate their antibacterial and enzyme inhibitory activities. Methods: E. milii plant extract was utilized for the green synthesis of AgNPs. The characterization of the NPs was performed through ultraviolet-visible spectroscopy (UV-Vis), Fourier-transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy (EDX). Antibacterial activity was assessed against Staphylococcus aureus, while enzyme inhibitory assays were conducted against urease, α-glucosidase, carbonic anhydrase II, and xanthine oxidase. Results: The synthesized AgNPs exhibited significant antibacterial effects, with a remarkable 20-mm zone of inhibition against S. aureus, surpassing the efficacy of the plant extract alone. Furthermore, the AgNPs demonstrated remarkable enzyme inhibition, achieving impressive percentages of 77.98% against α-glucosidase and 88.54% against carbonic anhydrase II. Half-maximal inhibitory concentration values for enzyme inhibition were highly promising, including 78.09 ± 1.98 µM for α-glucosidase, 0.22 ± 0.10 µM for carbonic anhydrase II, and 7.11 ± 0.55 µM for xanthine oxidase. Conclusion: In this study, AgNPs were successfully synthesized using E. milii extract and characterized using various techniques. The AgNPs exhibited significant antibacterial and enzyme-inhibitory activities, showcasing their potential for biomedical applications. [ABSTRACT FROM AUTHOR]

In the pursuit of identifying the novel resin glycoside modulators glucose-6-phosphatase and α-glucosidase enzymes, associated with blood sugar regulation, methanol-soluble extracts from the flowers of Ipomoea murucoides (cazahuate, Nahuatl), renowned for its abundance of glycolipids, were employed. The methanol-soluble extracts were fractionated by applying the affinity-directed method with glucose-6-phosphatase enzymes from a rat's liver and α-glucosidase enzymes from its intestines. Mass spectrometry and nuclear magnetic resonance were employed to identify the high-affinity compound as a free ligand following the release from the enzymatic complex. Gel permeation through a spin size-exclusion column allowed the separated high-affinity molecules to bind to glucose-6-phosphatase and α-glucosidase enzymes in solution, which led to the identification of some previously reported resin glycosides in the flowers of cazahuate, where a glycolipid mainly structurally related to murucoidin XIV was observed. In vitro studies demonstrated the modulating properties of resin glycosides on the glucose-6-phosphatase enzyme. Dynamic light scattering revealed conformational variations induced by resin glycosides on α-glucosidase enzyme, causing them to become more compact, akin to observations with the positive control, acarbose. These findings suggest that resin glycosides may serve as a potential source for phytotherapeutic agents with antihyperglycemic properties. [ABSTRACT FROM AUTHOR]

The industry has increasingly explored the development of foods with functional properties, where supplementation with probiotics and bioactive compounds has gained prominence. In this context, the study aimed to evaluate the influence of in vitro biological digestion on the content of phenolic compounds, antioxidant activity, and inhibition of α-amylase and α-glucosidase activities of probiotic yogurt supplemented with the lactic acid bacteria Lactococcus lactis R7 and red guava extract (Psidium cattleianum). A yogurt containing L. lactis R7 (0.1%) and red guava extract (4%) was characterized for the content of phenolic compounds, antioxidant activity, and potential for inhibition of digestive enzymes after a simulated in vitro digestion process. After digestion, the caffeic and hydroxybenzoic acids remained, and sinapic acid only in the last digestive phase. Antioxidant activity decreased during digestion by 28.93, 53.60, and 27.97% for DPPH, nitric oxide and hydroxyl radicals, respectively, and the inhibition of the α-amylase enzyme decreased only 4.01% after the digestion process. α-glucosidase was more efficient in intestinal digestion, demonstrating an increase of almost 50% in probiotic yogurt with red guava extract before digestion. Possibly, the phenolics change their conformation during digestion, generating new compounds, reducing antioxidant activity, and increasing the inhibitory activity of α-glucosidase digestive enzymes. It was concluded that the probiotic yogurt formulation supplemented with red guava extract could interfere with the concentration of phenolic compounds and the formation of new compounds, suggesting a positive and effective inhibition of the digestive enzymes, even after the digestive process. [ABSTRACT FROM AUTHOR]

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Four new glycosides, named amplexicosides A-D (1–4), and five known compounds: benzyl 2-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyloxy]-benzoate (5), benzyl 2-neohesperidosyloxy-6-hydroxybenzoate (6), chrysandroside A (7), chrysandroside B (8) and camelliquercetiside C (9) were isolated from the branches and leaves of Camellia amplexicaulis (Pit.) Cohen-Stuart. Their structures were elucidated using HR-ESI-MS and 1D- and 2D-NMR spectra and compared to reported NMR data. All of the isolated compounds were screened in an α-glucosidase assay. Compounds 4, 8, and 9 significantly inhibited α-glucosidase with respective IC50 values of 254.9 ± 4.2, 304.8 ± 11.9 and 228.1 ± 16.4 μM. [ABSTRACT FROM AUTHOR]

Debranching enzymes are unable to fully meet the requirements of green production of starch sugars and comprehensive utilization of by-products. Therefore, it is necessary to explore suitable new types of debranching enzymes. This study screened an oligo-1,6-glucosidase gene (oga) from Paenibacillus sp. STB16 and constructed its Bacillus subtilis expression system. This system achieved heterologous expression of the oligo-1,6-glucosidase. The enzymatic properties and substrate specificity of the recombinant oligo-1,6-glucosidase were explored. The results showed that the enzyme activity of the fermentation supernatant of B. subtilis could reach 162.33 U/mL. The recombinant oligo-1,6-glucosidase had the optimum reaction temperature at 50 °C and optimum reaction pH at 6.0. It specifically acted on the &#945-1, 6-glycosidic bonds of branches with degree of polymerization of 1. Compared to other types of debranching enzymes (substrate conversion rate under the detection limit), the recombinant oligo-1,6-glucosidase exhibited stronger hydrolytic activity towards isomaltose, isomaltotriose, and panose. The substrate conversion rate reach 97.4%-100%, with glucose as the product. Therefore, it is more suitable for treating glucose mother liquor. The glucose content in the first mother liquor, second mother liquor and tail liquor of chromatographic separation was significantly increased by 3.6%, 12.7% and 34.4%, respectively. The research results provide a theoretical basis for the development and utilization of new types of debranching enzymes, and also serve as an important reference for the comprehensive utilization of by-products in starch sugar processing. [ABSTRACT FROM AUTHOR]

Diabetes is often associated with increased oxidative stress caused by an imbalance between detoxification and ROS production. Unfortunately, many commercial drugs available today for treating this disease have adverse side effects and ultimately fail to restore glucose homeostasis. Therefore, finding a dietary anti-diabetic remedy that is safe, effective, and economical is crucial. In this study, GC–MS analysis, subsequent HPLC-assisted fractionation, and SPE-based purification led to identifying and purifying of key components such as phloroglucinol and total procyanidin dimer (procyanidin dimer and procyanidin dimer gallate) from methanolic seed extract of Vitis vinifera. In-vitro anti-diabetic screening of various fractions derived from methanolic extract along with individual components and their combinations revealed the potential synergistic behaviour of phloroglucinol and total procyanidin dimer with the lowest IC50 of 48.21 ± 3.54 µg/mL for α-glucosidase and 63.06 ± 5.38 µg/mL for α-amylase inhibition which is found to be superior to the effect shown by the standard Epigallocatechin gallate. Later Glucose utilization studies demonstrated the concentration-dependent effect of Phloroglucinol and total procyanidin dimer, and that has raised the glucose uptake by approximately 36–57% in HepG2 cells and 35–58% in L6 myocytes over a concentration of 50–100 µg/mL. The superior anti-diabetic effect of Phloroglucinol and total procyanidin dimer was proved by the suppression of oxidative stress with an IC50 of 7.92 ± 0.36 µg/mL for DPPH scavenging and 16.87 ± 1.24 µg/mL for SOD scavenging which is competent with the standard ascorbic acid. According to this study, suppressing ROS levels by phloroglucinol and total procyanidin dimer would be the underlying mechanism for the synergistic anti-diabetic effect of this combination. [ABSTRACT FROM AUTHOR]

The cellobiose-responsive regulator ClbR, a Zn(II)2Cys6 binuclear-cluster transcription factor, is a positive regulator of carbohydrate-active enzyme (CAZyme) genes responsive to cellulose in Aspergillus aculeatus. Because Zn(II)2Cys6 transcription factors tend to dimerize with proteins of the same family, we searched for a counterpart of ClbR and identified ClbR2, which is 42% identical to ClbR, as an interacting partner of ClbR by yeast two-hybrid screening. Genetic analyses suggested that ClbR and ClbR2 cooperatively regulate the expression of CAZyme genes in response to cellulose and 1,4-β-mannobiose in A. aculeatus. CAZyme genes under the control of the transcription factor ManR were regulated by ClbR and ClbR2, whereas those controlled by the transcription factor XlnR were regulated by ClbR, but not ClbR2. These findings suggest that ClbR participates in multiple regulatory pathways in A. aculeatus by altering an interacting factor. [ABSTRACT FROM AUTHOR]

The regulation of plant biomass degradation by fungi is critical to the carbon cycle, and applications in bioproducts and biocontrol. Trichoderma harzianum is an important plant biomass degrader, enzyme producer, and biocontrol agent, but few putative major transcriptional regulators have been deleted in this species. The T. harzianum ortholog of the transcriptional activator XYR1/XlnR/XLR-1 was deleted, and the mutant strains were analyzed through growth profiling, enzymatic activities, and transcriptomics on cellulose. From plate cultures, the Δxyr1 mutant had reduced growth on D-xylose, xylan, and cellulose, and from shake-flask cultures with cellulose, the Δxyr1 mutant had ~90% lower β-glucosidase activity, and no detectable β-xylosidase or cellulase activity. The comparison of the transcriptomes from 18 h shake-flask cultures on D-fructose, without a carbon source, and cellulose, showed major effects of XYR1 deletion whereby the Δxyr1 mutant on cellulose was transcriptionally most similar to the cultures without a carbon source. The cellulose induced 43 plant biomass-degrading CAZymes including xylanases as well as cellulases, and most of these had massively lower expression in the Δxyr1 mutant. The expression of a subset of carbon catabolic enzymes, other transcription factors, and sugar transporters was also lower in the Δxyr1 mutant on cellulose. In summary, T. harzianum XYR1 is the master regulator of cellulases and xylanases, as well as regulating carbon catabolic enzymes. [ABSTRACT FROM AUTHOR]

In response to the increasing prevalence of diabetes mellitus and the limitations associated with the current treatments, there is a growing need to develop novel medications for this disease. This study is focused on creating new compounds that exhibit a strong inhibition of alpha-glucosidase, which is a pivotal enzyme in diabetes control. A set of 33 triazole derivatives underwent an extensive QSAR analysis, aiming to identify the key factors influencing their inhibitory activity against α-glucosidase. Using the multiple linear regression (MLR) model, seven promising compounds were designed as potential drugs. Molecular docking and dynamics simulations were employed to shed light on the mode of interaction between the ligands and the target, and the stability of the obtained complexes. Furthermore, the pharmacokinetic properties of the designed compounds were assessed to predict their behavior in the human body. The binding free energy was also calculated using MMGBSA method and revealed favorable thermodynamic properties. The results highlighted three novel compounds with high biological activity, strong binding affinity to the target enzyme, and suitability for oral administration. These results offer interesting prospects for the development of effective and well-tolerated medications against diabetes mellitus. Dataset License: License under which the dataset is made available (CC0, CC-BY, CC-BY-SA, CC-BY-NC, etc.) [ABSTRACT FROM AUTHOR]