Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.2.1.21 (beta-glucosidase)
3,280 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Chemiluminescent assays of various enzymes have been developed using indoxyl derivatives as substrates. The principle of the method is as follows: an enzyme causes hydrolysis of an indoxyl derivative to an intermediate indoxyl that is readily oxidized to indigo dye and simultaneously produces hydrogen peroxide (H2O2). Hydrogen peroxide is detected chemiluminescently using isoluminol-microperoxidase. Alkaline phosphatase (ALP), beta-D-galactosidase (beta-gal), and beta-glucosidase were assayed by this method using 5-bromo-4-chloro-3-indolyl phosphate (BCIP), 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside (X-Gal), and 5-bromo-4-chloro-3-indolyl-beta-D-glucoside, respectively, as substrates. Using BCIP and X-Gal substrates, we have been able to detect 10(-19) mol of ALP and beta-gal, respectively. This assay system can be applied to enzyme immunoassay and DNA probe assay.
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PMID:Chemiluminescent assay of various enzymes using indoxyl derivatives as substrate and its applications to enzyme immunoassay and DNA probe assay. 172 50

Nine healthy volunteers were studied before, during, and after ingesting a fermented dairy product containing Lactobacillus acidophilus, Bifidobacterium bifidum, and mesophilic cultures (Streptococcus lactis and S cremoris) for 3 wk. Hydrogen and methane productions and fecal beta-galactosidase and beta-glucosidase activities were measured as indicators of fermentation capacity of the colonic flora. Fecal concentrations of nitroreductase, azoreductase, and beta-glucuronidase, which may be implicated in colonic carcinogenesis, were also assessed. Hydrogen and methane productions, fecal beta-galactosidase, beta-glucuronidase, and azoreductase activities did not change over three 3-wk periods whereas fecal beta-glucosidase activity increased (42 +/- 6, 91 +/- 12, and 40 +/- 6 IU/g N, P less than 0.01) and nitroreductase decreased (0.87 +/- 0.13, 0.54 +/- 0.11, and 0.57 +/- 0.08 IU/g N, P less than 0.05).
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PMID:Effect of chronic ingestion of a fermented dairy product containing Lactobacillus acidophilus and Bifidobacterium bifidum on metabolic activities of the colonic flora in humans. 211 57

Strained ruminal fluid was collected from cattle fed five diets at two locations to determine in vitro rates of cyanogenesis from the glycosides amygdalin, prunasin and linamarin. Rates of dissociation for the corresponding aglycones, benzaldehyde cyanohydrin and acetone cyanohydrin, also were determined. Hydrogen cyanide (HCN) in ruminal fluid was determined with a modified method of HCN analysis that independently measured the overall rate of cyanogenesis and the nonenzymatic dissociation of cyanohydrins, the intermediate products in the degradation of cyanogenic glycosides to HCN. Rate of dissociation of cyanohydrins in ruminal fluid was pH-dependent, with high rates of dissociation (as expressed by the rate constant or half-life of the reaction) occurring at pH greater than 6 and slower rates at pH 5 to 6. Cyanohydrin dissociation was most rapid when cattle were fasted for 24 to 48 h and ruminal pH was high; rate of dissociation was much slower during feeding and digestion. When the glycosides were examined, highest rates of cyanogenesis (mg HCN.liter-1.s-1) were observed after a 24-h postprandial period. Hence, cattle are most susceptible to poisoning by cyanogenic plants when the pH of ruminal fluid is elevated (for rapid dissociation) and also when the activity of microbial beta-glucosidase is adequate for rapid hydrolysis of glycosidic bonds. Rates of cyanogenesis were higher when ruminal inocula were from cattle fed fresh alfalfa or cubed alfalfa hay rather than grain or long hay. Rates of HCN production were slowest using inocula from cattle fed grain; rates for the three glycosides were negligible at the 3 and 6 h postprandial sampling times.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Factors that determine rates of cyanogenesis in bovine ruminal fluid in vitro. 216 29

Five anaerobic fungal isolates from the bovine rumen were grown on Coastal Bermuda grass (CBG) leaf blades and monitored over a 9-day period for substrate utilization, fermentation products, cellulase, and xylanase activities. Two of the fungal isolates showed monocentric growth patterns; one (isolate MC-1) had monoflagellated zoospores and morphologically resembled members of the genus Piromyces; the other (isolate MC-2) had multiflagellated zoospores and resembled members of the genus Neocallimastix. Three other isolates (PC-1, PC-2, and PC-3) exhibited polycentric growth and have not yet been described in the literature; these isolates were characterized by differences in morphology. All of the isolates degraded CBG to approximately the same extent (70% [dry weight]) in 9 days. Fermentation product accumulation was concurrent with substrate utilization. The major fermentation products for all isolates were formate, acetate, D-(-)-lactate, L-(+)-lactate, ethanol, carbon dioxide, and hydrogen. Succinate was produced by all cultures, with the exception of MC-1. Fermentation balances revealed different profiles for each isolate. As a group, monocentric isolates produced a greater ratio of oxidized to reduced products when grown on glucose or CBG than did the polycentric isolates, which produced a nearly equal ratio of these products. All isolates exhibited cellulolytic and xylanolytic activities, including endoglucanase, exoglucanase, beta-glucosidase, xylanase, and beta-xylosidase activities. Increasing enzyme activity correlated with the accumulation of fermentation products and substrate utilization. The optimum pH for the enzymatic activity of polycentric isolates was within a more narrow range (pH 6.4 to 7.0) than that of the monocentric isolates (pH 5.5 to 7.5). Activity toward cellulosic substrates was not detected until after the disappearance of reducing sugars. Xylanase activity was found to be five to seven times that of carboxymethyl cellulase activity for all cultures grown on CBG.
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PMID:Fermentation products and plant cell wall-degrading enzymes produced by monocentric and polycentric anaerobic ruminal fungi. 275 72

A beta-glucosidase has been purified from culture filtrates of the fungus Trichoderma reesei QM9414 grown on microcrystalline cellulose. The beta-glucosidase was purified using two successive DEAE-Sephadex anion-exchange chromatography steps, followed by SP-Sephadex cation-exchange chromatography and concanavalin-A--agarose chromatography. Evidence for homogeneity is provided by polyacrylamide disc gel electrophoretic patterns, which show a single protein band. Sedimentation equilibrium analysis yielded a molecular mass of 74.6 +/- 2.4 kDa. Sodium dodecyl sulfate/polyacrylamide gel electrophoresis yielded a single protein band with a molecular mass of 81.6 kDa. Thus, the enzyme appears to be a single, monomeric polypeptide. The beta-glucosidase is isoelectric at pH 8.5. The enzyme is rich in basic amino acids and contains few half-cystine and methionine residues. The purified beta-glucosidase contains less than 1% by weight of neutral carbohydrate. The beta-glucosidase catalyzes the hydrolysis of cellobiose, p-nitrophenyl beta-D-glucopyranoside and 4-methylumbelliferyl beta-D-glucopyranoside; the values of V/Km for each substrate were determined to be 2.3 X 10(4), 6.9 X 10(5) and 2.9 X 10(6) M-1 S-1 respectively. The enzyme is optimally active from pH 4.5 to 5.0 and is labile at higher hydrogen ion concentrations. The beta-glucosidase has an unusually high affinity for D-glucose (Ki = 700 microM). Comparison of inhibition constants for cello-oligosaccharides suggests that the substrate-binding region of the beta-glucosidase comprises multiple subsites.
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PMID:Purification and characterization of a beta-glucosidase from Trichoderma reesei. 310

The anaerobic rumen fungus Piromonas communis, when cultured on cotton fibre as the carbon source, produces an extracellular cellulase that is capable of solubilizing "crystalline" hydrogen-bond-ordered cellulose, in the form of the cotton fibre, at a rate that is greater than that of any other cellulases reported in the literature hitherto. The cell-free culture fluid is also very rich in xylan-degrading enzymes. The activity towards crystalline cellulose resides in a high-molecular-mass (approximately 700-1000 kDa) component (so-called crystalline-cellulose-solubilizing component, CCSC) that comprises endo (1-->4)-beta-D-glucanase (carboxymethylcellulase), beta-D-glucosidase and another enzyme that appears to be important for the breakdown of hydrogen-bond-ordered cellulose. The CCSC is associated with only a small amount of the endo(1-->4)-beta-D-glucanase (1.9%), beta-D-glucosidase (0.7%) and protein (0.5%) found in the crude cell-free cellulase preparation. The CCSC, unlike the bulk of the endo(1-->4)-beta-D-glucanase and beta-D-glucosidase, is very strongly absorbed on the microcrystalline cellulose, Avicel.
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PMID:Studies on the capacity of the cellulase of the anaerobic rumen fungus Piromonas communis P to degrade hydrogen bond-ordered cellulose. 763 4

Peroxidases (E.C. 1.11.1.7., hydrogen donor oxidoreductase) utilize hydrogen peroxide or substituted peroxides for the oxidation of a large number of substrates. Peroxidases are widely distributed and have been isolated from many higher plants (1). The wide distribution of the enzyme suggests that it could be of great biological importance, but the physiological functions and metabolic control of these enzymes are still poorly understood. The simultaneous presence of amine oxidase and peroxidase in cell walls suggests that the peroxide generated on oxidation of the amines could be utilized by the peroxidase (2,3). Recently we have purified an amine oxidase from Hordeum vulgare (4) and we have attempted to purify the peroxidase in order to study in vitro the reconstituted coupled system. beta-glucosidase (beta-D-glucoside glucohydrolase E.C. 3.2.1.21.) is capable of transforming glucosides in glucose and the corresponding aglycone or disaccharides as cellobiose, sophorose, gentiobiose. This enzyme is widely distributed in plants, fungi, bacteria, yeasts and animals (5,6). In the homogenate of Hordeum vulgare seedlings we also found beta-glucosidase activity and also attempted to purify beta-glucosidase. This enzyme copurified with peroxidase up to the last step. We report here the isolation of peroxidase and beta-glucosidase from Hordeum vulgare seedlings: some molecular and kinetic properties are given.
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PMID:Improved chromatographic purification of peroxidase and beta-glucosidase from Hordeum vulgare seedlings. 824 30

The role of noncovalent interactions in the catalytic mechanism of the Agrobacterium faecalis beta-glucosidase was investigated by steady-state and pre-steady state kinetic analysis of the hydrolysis of a series of monosubstituted aryl glycosides, in which the hydroxyl groups on the glycone were substituted by hydrogen or fluorine. Contributions of each hydroxyl group to binding of these substrates at the ground state are relatively weak (interaction energies of 3.3 kJ/mol or smaller) but are much greater at the two transition states (glycosylation and deglycosylation). The strongest transition state interactions were at the 2 position (at least 18 and 22 kJ/mol for glycosylation and deglycosylation, respectively) with the interactions at the 3 and 6 positions contributing at least another 9 kJ/mol of binding energy at both transition states. The interaction at the 4 position is less crucial to transition state binding but important for stabilization of the glycosyl-enzyme intermediate. Comparison of observed rates with those for spontaneous hydrolysis of the same substrates provides evidence for oxocarbenium ion character at both transition states, that for deglycosylation apparently having the greater positive charge development at the anomeric center.
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PMID:Mechanism of Agrobacterium beta-glucosidase: kinetic analysis of the role of noncovalent enzyme/substrate interactions. 851 77

The inhibition of four beta-glucosidases of plant, fungal, and mammalian origin by N1-butyl- and N1-dodecyl-D-gluconamidine was determined. Comparison with the inhibition by the corresponding N-alkyl-D-glucosylamines revealed that the strongly basic amidines (pKa 10.8) were at the most 10-times more inhibitory than the weakly basic glucosylamines (pKa 6.5). The small enhancement of inhibitory potency, resulting from transforming the tetrahedral C-1 geometry of the glucosylamines to the planar sp2-geometry of the amidines, was ascribed to the inability of the fully protonated amidines to function as hydrogen bond acceptors with the catalytic acid of the enzyme. Additional evidence for the importance of a hydrogen bond for strong inhibition came from the comparison of K1-values of the weakly basic 5-amino-5-deoxyhexopyranoses and 1,5-iminohexitols with those of the corresponding glyconamidrazones (pKa 8.4), which also have a planar C-1 geometry but are largely protonated under the assay conditions and which had similar or up to 10(4)-times larger K1-values than the former. Transition state resemblance was judged from the ratio KS(alkyl beta-glucoside)/K1(alkyl gluconamidine) relative to the rate acceleration factor kcat/kuncat (Wolfenden, Acc, Chem. Res., 5 (1972) 10-16). Compared to ratios of kcat/kuncat from > or = 10(11) to > or = 10(13), the ratios for KS/K1 were only from 10(3) to 2 x 10(4) except for beta-glucosidase A3 from Asp. wentii which had KS/K1 2.8 x 10(6). This enzyme differs from the others by being strongly inhibited by cationic glycon and substrate analogues rather than by basic ones. The pH-dependence of 1/K1 and the 'slow' approach to the inhibition is discussed with respect to transition state resemblance.
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PMID:N1-alkyl-D-gluconamidines: are they 'perfect' mimics of the first transition state of glucosidase action? 887 Feb 40

Recent work in the synthesis of cyclic phosphonate analogues of glucose [Darrow, J.W.; Drueckhammer, D.G. (1994) J. Org. Chem. 1994, 59, 2976] has been extended to the synthesis of a corresponding phosphonamidate analogue. A phosphonate salt, phosphonate methyl ester, and phosphonamidate analogue were tested as inhibitors of two inverting alpha-glycosidases, (trehalase and glucoamylase), and two retaining glycosidases, (alpha-glucosidase and beta-glucosidase). No inhibition of any of these enzymes was observed with the phosphonate salt or methyl ester. However, the phosphonamidate gave moderate competitive inhibition of the two inverting glycosidases and the retaining alpha-glucosidase but no inhibition of beta-glucosidase. The phosphonamidate showed enhanced binding relative to a simple monosaccharide only with the inverting glycosidases. This enhanced binding is believed to be due to hydrogen bonding interactions between the phosphonamidate group and two active site carboxylate residues implicated in catalysis. The selectivity toward inverting glycosidases is consistent with differences in distance of an active site carboxylate from the anomeric carbon of the glycoside substrate for the inverting versus the retaining glycosidases.
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PMID:A cyclic phosphonamidate analogue of glucose as a selective inhibitor of inverting glycosidases. 887 56


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