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Β-glucosidase (BGLs) act synergistically with endoglucanases and exoglucanases and then are of great interest for biomass conversion into bioethanol. Thus, the aim of the current study is to produce a recombinant β-glycosidase from Moniliophtora perniciosa expressed in Escherichia coli cells. Enzyme coding sequence expression was confirmed through Sanger sequencing after using wheat bran (WB) and carboxymethylcellulose (CMC) as fungal growth media. Synthetic gene betaglyc-GH1 with optimized codons for E. coli expression was cloned in pET-28a. β-glucosidase recombinant (GH1chimera) was purified using a nickel column and its identity was confirmed through mass spectrometry. The recombinant enzyme presented an apparent molecular mass of 53.23 kDa on SDS-PAGE. Recombinant β-glucosidase has shown hydrolytic activity using p-nitrophenyl-β-D-glycopyranoside (pNPG) as substrate and maximum activity at pH 4.6 and 65 °C. Thus, the results indicate that the application of the GH1chimera in the hydrolysis of lignocellulosic materials to obtain glucose monomers can be efficient. [ABSTRACT FROM AUTHOR]

Baicalein and wogonin, two health-promoting flavonoid aglycones in Scutellaria baicalensis Georgi roots, have better bioactivity and bioavailability than their glycoside precursors (baicalin and wogonoside). The search for novel microorganisms for the fermentation of S. baicalensis roots to improve the yields of baicalein and wogonin is of great interest in the food and nutraceutical industries. In this study, a novel endophytic fungus (Penicillium rubens) capable of producing cellulase (0.82 ± 0.12 U/mL) and β-glucosidase (4.55 ± 0.97 U/mL) was used to ferment S. baicalensis roots to obtain high yields of baicalein and wogonin. Semi-solid-state fermentation with homogeneous fungal cultures was found to be the appropriate strategy. Under the optimal fermentation parameters (temperature 30 °C, inoculation dose of fungal cultures 4 mL/g, and time 60 h), the yields of baicalein and wogonin (46.95 ± 4.98 mg/g DW and 31.41 ± 9.36 mg/g DW) from S. baicalensis roots were tremendously increased by 16.13-fold and 15.47-fold over control, respectively. Results of industrial enzyme bioprocessing and scanning electron microscope indicated that the remarkable increase in aglycone product yields could be due to the synergistic effects of cellulase and β-glucosidase produced by P. rubens on cell wall hydrolysis and glycoside deglycosylation. In addition, extracts of S. baicalensis roots fermented by P. rubens showed a significant increase in antioxidant and antimicrobial activities. Overall, the semi-solid-state fermentation of S. baicalensis roots using the novel P. rubens was an effective strategy that could yield high quantities of baicalein and wogonin, which had promising potential in the food and nutraceutical industries. [ABSTRACT FROM AUTHOR]

A study was conducted to investigate the anti-diabetic and anti-urease potential of Osbeckia nutans leaves (ONL). Six compounds, i.e. quercetin-3-O-glucoside, myricetin, shikimic acid, catechin, trans-ferulic acid and luteolin were identified from the butanol sub-fraction, BE2 and the ethyl acetate sub-fraction, EA5 of ONL. BE2 inhibited α-glucosidase and Jack bean urease with IC50 values of 0.036 μg/mL (437.46 μg/mL for acarbose) and 0.327 mg/mL (0.039 mg/mL for thiourea), respectively. In the glucose uptake experiment, BE2 (0.05 mg/mL) treatment resulted in a substantial increase in glucose uptake in free fatty acid (FFA)-treated cells at a concentration 10 times lower than that seen in EA5 (0.5 mg/mL) treated cells. The binding energies of quercetin-3-O-glucoside with α-glucosidase, glucose transporter GLUT4 and H. pylori urease were found to be −94.2585, −219.8271 and −254.391 kcal/mol, respectively. This study revealed that ONL has anti-diabetic and anti-urease abilities and further in-depth research may unveil its full potential. [ABSTRACT FROM AUTHOR]

Abstractβ-D-xylosidase, an important glycoside hydrolase, hydrolyzes xylo-oligosaccharides to yield xylose and biotransforms specific saponins by cleaving β-xylose at the non-reducing end. It is extensively employed in renewable resources, including fuels, food, and pharmaceuticals, as a xylanolytic enzyme. In this study, a 1197 bp β-xylosidase gene (BaXyl5B) derived from Bifidobacterium adolescentis was cloned and expressed in E. coli BL21. Following purification, the recombinant β-xylosidase exhibited a distinct band on SDS-PAGE; its molecular weight was determined to be 46.4 kDa. The optimal activity of BaXyl5B was observed at a pH of 6.0 and a temperature of 30 °C. Purified BaXyl5B demonstrated multifunctional activities involving p-nitrophenyl-β-D-xylopyranoside (pNPβXyl), p-nitrophenyl-α-L-arabinopyranoside (pNPαArap), and p-nitrophenyl-β-D-glucopyranoside (pNPβGlc). A significant finding of this study was that BaXyl5B exhibited high selectivity towards ginsenoside Rb3, which could hydrolyze xylose at the C-20 position of Rb3 to yield Rd and F2. Overall, this research explored a novel β-xylosidase, BaXyl5B, that could serve as a valuable biocatalyst for converting ginsenosides and synthesizing active glycosides and aglycones, making it a promising enzyme for further exploration and utilization. [ABSTRACT FROM AUTHOR]

Type-2 diabetes affects an increasing percentage of the world's population and its control through dietary management, involving the consumption of health-promoting foods or their derived supplements, is a common strategy. Several mushroom species have been demonstrated to be endowed with antidiabetic properties, resulting from their ability in improving insulin sensitivity and production, or inhibiting the carbohydrate-hydrolyzing enzymes α-amylase and α-glucosidase. This study aimed to investigate for the first time the hypoglycemic properties of the edible mushroom Leccinum scabrum (Bull.) Gray. Mushroom extracts were prepared through the microwave-assisted extraction (MAE) technique using green solvents with different polarity degrees. The inhibition activity of all the obtained extracts on both α-glucosidase and α-amylase was evaluated and the highest activity was observed for the EtOAc extract which showed an IC50 value about 60-fold lower than the reference compound 1-deoxynojirimycin (DNJ) on α-glucosidase (0.42 ± 0.02 and 25.4 ± 0.6 µg/mL, respectively). As expected on the basis of the literature data concerning both α-glucosidase and α-amylase inhibition, a milder inhibition activity on pancreatic α-amylase was observed. Preliminary in vivo tests on Drosophila melanogaster carried out on the most active obtained extract (EtOAc) confirmed the in vitro observed hypoglycemic activity. Finally, the EtOAc extract metabolic profile was determined through GC-MS and HRMS analyses. [ABSTRACT FROM AUTHOR]

β‐Glucosidase from the thermophilic bacterium Caldicellulosiruptor saccharolyticus (Bgl1) has been denoted as having an attractive catalytic profile for various industrial applications. Bgl1 catalyses the final step of in the decomposition of cellulose, an unbranched glucose polymer that has attracted the attention of researchers in recent years as it is the most abundant renewable source of reduced carbon in the biosphere. With the aim of enhancing the thermostability of Bgl1 for a broad spectrum of biotechnological processes, it has been subjected to structural studies. Crystal structures of Bgl1 and its complex with glucose were determined at 1.47 and 1.95 Å resolution, respectively. Bgl1 is a member of glycosyl hydrolase family 1 (GH1 superfamily, EC 3.2.1.21) and the results showed that the 3D structure of Bgl1 follows the overall architecture of the GH1 family, with a classical (β/α)8 TIM‐barrel fold. Comparisons of Bgl1 with sequence or structural homologues of β‐glucosidase reveal quite similar structures but also unique structural features in Bgl1 with plausible functional roles. [ABSTRACT FROM AUTHOR]

The upwelling phenomenon plays a vital role within marine ecosystems, transporting essential nutrients from the bottom to the surface and boosting biological productivity. However, the bacterial community structure in upwelling zones along the western coast of India (WCI) is understudied. This research systematically examines bacterial diversity across three seasons—pre-monsoon (PR), monsoon (MN), and post-monsoon (PM)—using next-generation sequencing. Our findings show distinct spatial patterns of bacterial communities in the Arabian Sea and demonstrate that ecological variations influence bacterial distribution in this dynamic environment. During MN, the bacterial community exhibited greater species diversity but lower overall abundance compared to PR and PM. Non-Metric MDS cluster analysis revealed a 78% similarity (at order level) between PR and PM, indicating that MN supports unique bacterial diversity. KEGG analysis showed significant seasonal variations in metabolic functions, with increased functional potential during MN. Additionally, Carbohydrate-Active enZymes (CAZymes) analysis revealed distinct seasonal profiles, among which the GH13 enzymes were the most prevalent glycoside hydrolases during MN, predominantly being sucrose phosphorylase and glucosidase, known for breaking down glucan deposits derived from phytoplankton. The CAZymes profiles supported taxonomic and KEGG pathway findings, reinforcing that microbial communities are seasonally distinct and functionally adapted to changing availability of nutrients. [ABSTRACT FROM AUTHOR]

The glycoside hydrolase family 3 (GH3) β-glucosidases from filamentous fungi are crucial industrial enzymes facilitating the complete degradation of lignocellulose, by converting cello-oligosaccharides and cellobiose into glucose. Understanding the diverse domain organization is essential for elucidating their biological roles and potential biotechnological applications. This research delves into the variability of domain organization within GH3 β-glucosidases. Two distinct configurations were identified in fungal GH3 β-glucosidases, one comprising solely the GH3 catalytic domain, and another incorporating the GH3 domain with a C-terminal fibronectin type III (Fn3) domain. Notably, Streptomyces filamentous bacteria showcased a separate clade of GH3 proteins linking the GH3 domain to a carbohydrate binding module from family 2 (CBM2). As a first step to be able to explore the role of accessory domains in β-glucosidase activity, a screening system utilizing the well-characterised Aspergillus niger β-glucosidase gene (bglA) in bglA deletion mutant host was developed. Based on this screening system, reintroducing the native GH3-Fn3 gene successfully expressed the gene allowing detection of the protein using different enzymatic assays. Further investigation into the role of the accessory domains in GH3 family proteins, including those from Streptomyces, will be required to design improved chimeric β-glucosidases enzymes for industrial application. [ABSTRACT FROM AUTHOR]

AbstractAlpha glucosidases are multifunctional glycoside hydrolases with hydrolysis and transglycosylation ability. It can be utilized for the glycosidic bond synthesis or glycosylation of Ascorbic acid/Vitamin C to its stable analogue, Ascorbic acid 2 glucoside (AA2G), a compound with wide applications in cosmetics and pharma. The application of α-glucosidases for industrial scale transglycosylation is limited due to the low transglycosylation yield of free enzymes. Enzyme immobilization techniques could enable the development of efficient, reusable catalysts. Only a few glycoside hydrolases have been studied in immobilized form for transglycosylation reactions, and α-glucosidases are probably the least explored in this form. Transglycosylation activity of immobilized α-glucosidase from Aspergillus carbonarius BTCF 5 was studied for AA2G synthesis, where different immobilization techniques like calcium alginate encapsulation, adsorption on chitosan beads, covalent cross-linking on magnetic nanoparticles, and cross-linked enzyme aggregates (CLEA) were employed for the immobilization. The immobilization yield of calcium alginate encapsulated enzyme, enzyme immobilized on Fe-MNP support, enzyme immobilized on chitosan beads and as CLEA were 107%, 99%, 46% and 486%, respectively. CLEA was identified as the best immobilization technique for this bi-substrate reaction due to the high immobilization yield and activity retention (30% activity retained after 5 consecutive cycles). Enzyme immobilization increased the transglycosylation activity by 38%, yielding 118 mM AA2G against 72 mM by the free enzyme. This indicates the potential of immobilized α-glucosidase as a catalyst for synthesizing AA2G at an industrial scale. [ABSTRACT FROM AUTHOR]

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]

β-glucosidase is a key component of cellulase for its function in hydrolyzing cellobiose to glucose in the final step of cellulose degradation. The high-level expression of β-glucosidase is essential for cellulose conversion. Aspergillus niger ATCC 20611 has the potential for efficient protein expression because of its ability to secret enzymes for the industrial production of fructooligosaccharides, but it lacks robust promoters for high-level protein expression. Here, the development of A. niger 20611 as a powerful protein expression system exploited the conserved constitutive promoter Pgpd1 of the glyceraldehyde-3-phosphate dehydrogenase-encoding gene from Trichoerma reesei to drive the expression of the enhanced green fluorescent protein in A. niger ATCC 20611. The mycelium of the transformant AGE9 exhibited intense fluorescence. Then, the promotor Pgpd1 was used to drive the expression of β-glucosidase and the enzyme activity of transformants AGB1 and AGB33 were 1.02 and 0.51 U/mL, respectively. These results demonstrate that the promotor Pgpd1 from T. reesei was applicable for A. niger ATCC 20611. Furthermore, the T. reesei-specific robust promoter Pcdna1 was used to drive the expression of β-glucosidase. The β-glucosidase exhibited a high-level expression with a yield of 15.2 U/mL, which was over 13.9 times higher than that driven by the promoter Pgpd1. The β-glucosidase was thermally stable and accounted for 85% of the total extracellular proteins. Subsequently, the fermentation broth including β-glucosidase was directly added to the cellulase mixture of T. reesei for saccharification of the acid-treated corncob residues and the delignified corncob residues, which increased the saccharification efficiency by 26.21% and 29.51%, respectively. Thus, β-glucosidase exhibited a high level of expression in A. niger ATCC 20611 and enhanced cellulose degradation by addition in vitro. In addition, the robust promoter Pcdna1 of T. reesei could drive the high-level expression of protein in A. niger ATCC 20611. These results demonstrate that the promoters in filamentous fungi could be employed across species in A. niger ATCC 20611 and further facilitated the efficient expression of β-glucosidase to optimize cellulases for efficient cellulose transformation. [ABSTRACT FROM AUTHOR]

Pholidota cantonensis Rolfe is an edible medicinal plant in the genus Pholidota of the family Orchidaceae. This plant is used to prepare medicated food in China and has been reported to possess anti-α-glucosidase activity. To date, little is known about the active substances responsible for the observed anti-α-glucosidase activity. In the present study, we aimed to screen and characterize the α-glucosidase inhibitory fraction of P. cantonensis using ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC–Q–TOF–MS/MS) analysis and molecular docking. As a result, the 50% ethanol fraction obtained from D101 macroporous adsorption resin column chromatography (D50 fraction) had the highest total phenol content (353.83 ± 6.06 mg GAE/g) and the most prominent α-glucosidase inhibitory activity (IC50 = 30.01 ± 7.30 µg/mL). Forty-five compounds were identified from the D50 fraction by using UPLC–Q–TOF–MS/MS analysis. Molecular docking results showed that six main constituents, namely, crepidatin, 2,7-dihydroxy-4-methoxyl-9,10-dihydrophenylene, 4,4ʹ,5,6-tetrahydroxystilbene, 4,7-dihydroxy-2-methoxyl-9,10-dihydrophenylene, (-)-lariciresinol, and thunalbene, in the D50 fraction occupied the catalytic sites of α-glucosidase through strong hydrophobic interactions, hydrogen bonding, and other patterns. The binding energies were between − 29.95 and − 11.41 kJ/mol, indicating good binding between the tested compounds and α-glucosidase. The active ingredients responsible for the α-glucosidase inhibitory activity may include phenanthrenes, stilbenes, dibenzyls, and lignans. The D50 fraction has potential value for developing innovative drugs for the prevention and treatment of diabetes mellitus (DM) and is worthy of in-depth research. [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]

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]

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]

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]

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]

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]

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]