Glucosidases are an indispensable class of enzymes in the sugar metabolism of organisms. To investigate the biological functions and expression patterns of α-glucosidases (AGLUs) and β-glucosidases (BGLUs), we identified the two family members in the genome of melon (Cucumis melo). The number, location on chromosomes, gene structure, subcellular localization, conserved motifs, and phylogenetic relationship of the two family members were analyzed. Based on the cis-acting elements in the promoter region and protein interaction models, their functions were preliminarily predicted. Furthermore, the gene expression of the two family members was determined by qRT-PCR. The results showed that the melon genome contained five AGLU family members on five chromosomes, and all of the five members were located in the extracellular matrix, with the amino acid sequence lengths ranging from 899 aa to 1 060 aa. The melon genome carried 18 BGLU family members on 8 chromosomes, and all the members were located in the cell membrane or cytoplasm, with the amino acid lengths ranging from 151 aa to 576 aa. The qRT-PCR results showed that the expression of about 50% of the genes was down-regulated upon cold stress. CmAGLU5 and CmBGLU7 may be key members of the two families, respectively, in response to cold stress. The expression of all members of the two families was up-regulated under abscisic acid (ABA), high salt, and drought stress. In the AGLU family, CmAGLU3 was the key gene in response to ABA and high salt stress, while CmAGLU4 was the key gene in response to drought stress. In the BGLU family, CmBGLU18 was the key gene in response to ABA, while CmBGLU6 was the key gene in response to high salt and drought stress.
beta-Glucosidase/metabolism* ; Phylogeny ; alpha-Glucosidases/metabolism* ; Gene Expression Regulation, Plant ; Cucurbitaceae/enzymology* ; Multigene Family ; Cucumis melo/enzymology* ; Stress, Physiological

Sugarcane (Saccharum spp.) is an important sugar crop. Biotic and abiotic stresses such as diseases, cold, and drought are major factors limiting sugarcane production. β-1,3-glucanase (EC 3.2.1.39), a member of the pathogenesis-related protein family, plays an essential role not only in the plant defenses against pathogens but also in plant growth, development, and abiotic stress responses. To systematically investigate the sugarcane β-1,3-glucanase gene family, 132 glycoside hydrolase (GH) 17 family members were identified in the genomes of the sugarcane wild species Saccharum spontaneum 'Np-X', the tropical species S. officinarum 'LA-Purple', and the Saccharum spp. hybrid cultivar 'R570'. The results of the phylogenetic analysis categorized them into four subfamilies, of which subfamily Ⅳ had the largest proportion of members (102). The members of the sugarcane GH17 gene family contained five conserved motifs and 0-16 introns. The majority of the GH17 genes exhibited a genome-wide replication pattern, with 89.50% originating from S. spontaneum 'Np-X' and S. officinarum 'LA-Purple', while 58.10% of them in the Saccharum spp. hybrid cultivar 'R570' belonged to the discrete replication type. Four major classes of cis-acting elements were identified in the promoters, including the elements related to plant growth, development, and tissue-specific expression (14.21%), light-responsive elements (38.24%), biotic or abiotic stress-responsive elements (9.18%), and hormone-responsive elements (38.37%), which suggested that this gene family was involved in plant growth, development, hormone responses, and stress responses. Transcriptome and quantitative real-time PCR (RT-qPCR) analyses showed that the sugarcane GH17 genes exhibited tissue-specific expression and were differentially expressed under low temperature, drought, and hormone treatments, as well as during the interactions between different sugarcane genotypes and Sporisorium scitamineum, suggesting their potential roles in plant defenses. In addition, some SsGlu genes (SsGlu5, SsGlu20, SsGlu21, SsGlu25, SsGlu28, and SsGlu39) were expected to serve as candidate stress-related genes. This study lays a foundation for further revealing the molecular mechanisms of the stress resistance of sugarcane via β-1,3-glucanase genes.
Saccharum/physiology* ; Stress, Physiological/genetics* ; Glucan 1,3-beta-Glucosidase/metabolism* ; Multigene Family ; Phylogeny ; Gene Expression Regulation, Plant ; Plant Proteins/genetics*

Six new prenylated flavonoid glycosides, including four new furan-flavonoid glycosides wushepimedoside A-D (1-4) and two new prenyl flavonoid derivatives wushepimedoside E-F (5-6), and one know analog epimedkoreside B (7) were isolated from biotransformation products of the aerial parts of Epimedium wushanense. Their structures were elucidated according to comprehensive analysis of HR-MS and NMR spectroscopic data, and the absolute configurations were assigned using experimental and calculated electronic circular dichroism (ECD) data. The regulatory activity of compounds 1-7 on the production of testosterone in primary rat Leydig cells were investigated, and 4 and 5 exhibited testosterone production-promoting activities. Molecular docking analysis suggested that bioactive compounds 4 and 5 showed the stable binding with 3β-HSD and 4 also had good affinity with Cyp17A1, which suggested that these compounds may regulate testosterone production through stimulating the expression of the above two key proteins.
Animals ; Epimedium/chemistry* ; Flavonoids/chemistry* ; Furans ; Glycosides/chemistry* ; Hydrolysis ; Male ; Molecular Docking Simulation ; Molecular Structure ; Rats ; Testosterone ; beta-Glucosidase/metabolism*

β-glucosidase has important applications in food, medicine, biomass conversion and other fields. Therefore, exploring β-glucosidase with strong stability and excellent properties is a research hotspot. In this study, a GH3 family β-glucosidase gene named Iubgl3 was successfully cloned from Infirmifilum uzonense. Sequence analysis showed that the full length of Iubgl3 was 2 106 bp, encoding 702 amino acids, with a theoretical molecular weight of 77.0 kDa. The gene was cloned and expressed in E. coli and the enzymatic properties of purified IuBgl3 were studied. The results showed that the optimal pH and temperature for pNPG hydrolysis were 5.0 and 85 ℃, respectively. The enzyme has good thermal stability, and more than 85% of enzyme activity can be retained after being treated at 80 ℃ for2 h. This enzyme has good pH stability and more than 85% of its activity can be retained after being treated at pH 4.0-11.0 for 1 h. It was found that the enzyme had high hydrolysis ability to p-nitrophenyl β-d-glucoside (pNPG) and p-nitrophenyl β-d-xylopyranoside (pNPX). When pNPG was used as the substrate, the kinetic parameters K_m and V_max were 0.38 mmol and 248.55 μmol/(mg·min), respectively, and the catalytic efficiency k_cat/K_m was 6 149.20 s^-1mmol^-1. Most metal ions had no significant effect on the enzyme activity of IuBgl3. SDS completely inactivated the enzyme, while EDTA increased the enzyme activity by 30%. This study expanded the β-glucosidase gene diversity of the thermophilic archaea GH3 family and obtained a thermostable acid bifunctional enzyme with good industrial application potential.
beta-Glucosidase/chemistry* ; Archaea/metabolism* ; Escherichia coli/metabolism* ; Hydrogen-Ion Concentration ; Temperature ; Glucosides ; Enzyme Stability ; Substrate Specificity ; Kinetics

This study aims to observe different factors which affected the bionic enzymatic hydrolysis of icariin into baohuoside I and to optimize the reaction conditions in order to provide research foundation for building a novel bionic enzymolysis drug delivery system. To simulate the environment in vivo, 37 degrees C was set as the temperature and artificial intestinal juice and gastric juice were selected as the buffer solutions. Taking the conversion of baohuoside I as index, the effects of the kinds of enzyme, enzyme activity, substrate concentration, reaction time, pancreatin in artificial intestinal juice and surfactant on the conversion of baohuoside I were investigated. The results showed that cellulase, beta-glucosidase and snailase were all inactive in artificial gastric juice and no baohuoside I generated. Pancreatin in artificial intestinal juice couldn't significantly influence the activity of beta-glucosidase or snailase (P > 0.05), but noticeably decrease the activity of cellulase (P < 0.05). In artificial intestinal juice, the conversion of baohuoside I was highest by using beta-glucosidase, and the optimum reaction conditions were determined as follows: enzyme activity 10 U x mL(-1), substrate concentration 1 mg x mL(-1), 3 g x L(-1) rhamnolipid and reaction time 3 h. Under this condition, the conversion of baohuoside I was 99.8%.
Animals ; Cellulase ; chemistry ; Flavonoids ; biosynthesis ; metabolism ; Hydrolases ; chemistry ; isolation & purification ; Hydrolysis ; Pancreatin ; chemistry ; Snails ; enzymology ; Surface-Active Agents ; chemistry ; beta-Glucosidase ; chemistry

Constructing ethanologenic strains with cellulose activity is important to achieve consolidated bioprocessing of lignocellulose for ethanol production. In this study, we integrated the pyruvate decarboxylase gene pdc and alcohol dehydrogenase gene adhB from Zymomonas mobilis ZM4 into Escherichia coli JM109 by Red recombination method to generatea recombinant strain E. coli P81 that could produce ethanol from glucose. Abeta-glucosidase gene bglB from Bacillus polymyxa 1.794 was cloned into the recombinant E. coli P81 and beta-glucosidase was expressed to give a new recombinant strain E. coli P81 (pUC19-bglB) with dual functions of cellobiose degradation and ethanol production. The extracellular beta-glucosidaseactivity was 84.78 mU/mL broth and the extracellular cellobiase activity of E. coli P81 (pUC19-bglB) was 32.32 mU/mL broth. E. coli P81 (pUC19-bglB) fermented cellobiose to ethanol with a yield of 55.8% of the theoretical value, and when glucose and cellobiose were co-fermented, the ethanol yield reached 46.5% of thetheoretical value. The construction of consolidated bioprocessing strain opens the possibility to convert cellobiose to ethanol in a single bioprocess.
Bacterial Secretion Systems ; Cellulose ; metabolism ; Escherichia coli ; genetics ; metabolism ; Ethanol ; metabolism ; Fermentation ; Recombinant Proteins ; biosynthesis ; genetics ; beta-Glucosidase ; biosynthesis ; genetics

Exploring new beta-glucosidase genes is of great importance to industrialize beta-glucosidase. The genomes of Aspergillus fumigatus contain a bgl gene, which encodes a 65 kDa putative beta-glucosidase. The bgl gene was cloned into an expression plasmid and transformed to Escherichia coli BL21 (DE3). The bgl was expressed upon induction of Isopropyl beta-D-1-thiogalactopyranoside (IPTG). The recombinant protein was purified by GST-tag affinity chromatography. The purified recombinant Bgl was characterized using Esculin as substrate. The optimum temperature and pH were 45 degrees C and 5.0-6.0, respectively. The K(m) for Esculin was 17.7 mmol/L. The enzyme was stable in the range of pH 4-7. After incubation at 70 degrees C for 2 h, the recombinant Bgl remained 60% of its activity. Metal ions and chemical reagents had different influences on the activity of beta-glucosidase. Ca2+ (1 mmol/L) could increase enzyme activity slightly. On the contrary, the enzyme activity was greatly inhibited by 5 mmol/L Sodium dodecyl sulfate (SDS). Based on our results, the A. fumigatus Bgl was thermostable beta-glucosidase.
Aspergillus fumigatus ; enzymology ; Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; beta-Glucosidase ; biosynthesis ; genetics ; metabolism

In order to explore the influence of reaction temperature on the product composition, the effect of continuous temperature change (22 degrees C-60 degrees C, +/-0.1 degree C) on hydrolysis of yeast beta-glucan by endo-beta-1,3-glucanase was determined by using self-developed Biochem-temperature Characteristic Apparatus. The activation energy of enzymatic hydrolysis of yeast beta-glucan was 84.17 kJ/mol. The optimum temperature represented by accumulation of products decreased exponentially within a certain period of time. The components of the products were changed with reaction temperature. The length of oligosaccharides decreased with the increase of temperature. The main products were laminaribiose and laminaritriose at the temperature higher than 46 degrees C, while the main products were laminaripentaose and larger molecular weight components at the temperature lower than 30 degrees C. The results can provide precise parameters to control the reaction temperature of the production of 1,3-beta-D-glucooligosaccharides.
Enzyme Activation ; Glucan Endo-1,3-beta-D-Glucosidase ; chemistry ; metabolism ; Hydrolysis ; Oligosaccharides ; chemistry ; metabolism ; Temperature ; Yeasts ; metabolism ; beta-Glucans ; metabolism

The interaction between genistein and beta-glucosidase was studied using fluorescence quenching method and synchronous fluorimetry. The binding reaction was simultaneously studied by the AutoDock 4.2 molecular docking model. Data from fluorescence spectroscopy indicated that these interactions resulted in the endogenous fluorescence quenching of beta-glucosidase, which belongs to a static quenching mechanism. The calculated binding constants were 3.69 x 10(4), 3.06 x 10(4) and 2.36 x 10(4) L x mol(-1) at 17, 27 and 37 degrees C, respectively. The evidences from synchronous fluorescence showed the effect of genistein on the microenvironment around beta-glucosidase in aqueous solution. The inhibition test showed that the activity of beta-glucosidase could be inhibited by genistein. The determined bimolecular rate constant (k(i)) was 1.2 x 10(3) (mol x L(-1)(-1) x min(-1). Molecular docking was performed to reveal the possible binding mode or mechanism and suggested that genistein could bind strongly to beta-glucosidase. The results revealed that genistein tended to bind with beta-glucosidase mainly by hydrogen bond and hydrophobic interaction as well as electrostatic forces.
Genistein ; metabolism ; Molecular Docking Simulation ; Protein Binding ; Protein Kinase Inhibitors ; metabolism ; Spectrometry, Fluorescence ; beta-Glucosidase ; metabolism

OBJECTIVEThis study was conducted to compare serum cytosolic &beta;-glucosidase (CBG) levels of age-matched control patients with those of infants with necrotizing enterocolitis (NEC) thereby to determine the eventual association between serum CBG levels with extensive disease in infants with NEC.

METHODA total of 96 premature infants were divided into the early NEC group (n = 25), confirmed NEC group (n = 23) and the control group (n = 48). Serum CBG concentration, C-reactive protein (CRP) and peripheral blood white blood cells (WBC) were measured at the onset of the disease in patients in early NEC or confirmed NEC groups and at weeks 2-3 in control infants. Data were analyzed using descriptive statistics, non-parametric tests, Student's t-test, linear correlation, Spearman correlation analysis, receiver operating characteristic (ROC) curve were used for statistical analysis.

RESULTSThe median birth weights (mean ± SE) in the three groups were not statistically significant (P > 0.05). Serum CBG concentration in the 3 groups were (112.369 ± 108.539) nmol/L, (693.013 ± 211.614) nmol/L and (36.478 ± 28.31) nmol/L, respectively. The infants in the confirmed NEC group had highest CBG levels, compared with the other 2 groups (P < 0.05). When the levels of CBG ≥ 65 ng/ml, CRP ≥ 2 mg/L and WBC < 5 × 10(9)/L within 3 days after birth or > 20 × 10(9)/L 3 days after birth were considered as positive parameters, the sensitivity of CBG and CRP was higher than that of WBC (P < 0.05). Among these indices, CBG had the highest specificity (87.4%), positive predictive (95.6%) and Youden's index (81.3%). CBG is correlated with CRP (the Spearman correlation coefficient was 0.379, P < 0.01).

CONCLUSIONSerum CBG concentration increases early in NEC. Serum CBG level was associated with extensive disease in infants with NEC. Therefore CBG can be used as a marker in the early diagnosis of NEC.

Case-Control Studies ; Enterocolitis, Necrotizing ; blood ; diagnosis ; Humans ; Infant, Newborn ; Infant, Premature ; Leukocyte Count ; Serum ; metabolism ; beta-Glucosidase ; blood