scholarly journals Functional lysosomal hydrolase size as determined by radiation inactivation analysis

1985 ◽  
Vol 226 (1) ◽  
pp. 283-288 ◽  
Author(s):  
G Dawson ◽  
J C Ellory
Keyword(s):  
Target Size ◽  
Lysosomal Hydrolase ◽  
Lysosomal Hydrolases ◽  
Range Analysis ◽  
Freeze Dried ◽  
Arylsulphatase A ◽  
Alpha Galactosidase ◽  
Beta Glucosidase ◽  
Alpha Glucosidase ◽  
Beta Galactosidase

Electron inactivation analysis with 16 MeV electrons was used to determine the functional target size of a number of commonly studied lysosomal hydrolases. Observed values ranged from a low of 62 000 +/- 4000 Da for beta-galactosidase to a high of 200 000 +/- 17 500 Da (mouse beta-glucuronidase). One group of lysosomal hydrolases (N-acetyl-beta-glucosaminidase, N-acetyl-beta-galactosaminidase, alpha-galactosidase, beta-mannosidase, beta-glucosidase, arylsulphatase A and sphingomyelinase) had target sizes in the range 100 000-120 000 Da, whereas alpha-glucosidase and alpha-fucosidase exist as complex multimers in the 150 000-160 000 Da range. Analysis of freeze-dried cell material showed little evidence of species (mouse versus human) variation in the functional size of most lysosomal hydrolases with the exception of beta-glucuronidase. Our findings suggest the potential usefulness of lysosomal hydrolases as endogenous marker enzymes in studies where the target size of proteins of unknown molecular mass is to be determined.

scholarly journals Soil hydrolase activities and kinetic properties as affected by wheat cropping systems of Northeastern China

2010 ◽  
Vol 56 (No. 11) ◽  
pp. 526-532 ◽  
Author(s):  
Y.L. Zhang ◽  
L.J. Chen ◽  
C.X. Sun ◽  
Z.J. Wu ◽  
Z.H. Chen ◽  
...  
Keyword(s):  
Cropping Systems ◽  
Reaction Rates ◽  
Total Carbon ◽  
Kinetic Properties ◽  
Total N ◽  
Alpha Galactosidase ◽  
Beta Glucosidase ◽  
Alpha Glucosidase ◽  
Beta Galactosidase ◽  
Sequential Cropping

Agricultural practices that reduce soil degradation and improve agriculture sustainability are important particularly for dry hilly land of Chaoyang County in the Liaoning Province, North-east China, where cinnamon soils are widely distributed and mainly for wheat production. The impacts of 10-year cropping systems (wheat-cabbage sequential cropping, wheat-corn intercrop, wheat-sunflower rotation, wheat-soybean rotation) on soil enzyme properties of surface-soil (0&ndash;20 cm) were studied. Total carbon, nitrogen, phosphorus and sulfur, and nine soil hydrolases related to nutrient availabilities (&beta;-galactosidase, &alpha;-galactosidase, &beta;-glucosidase, &alpha;-glucosidase, urease, protease, phosphomonoesterase, phosphodiesterase, arylsulphatase) and five enzymes kinetic characters were examined. Wheat-corn intercrop systems had higher total C, total N, total P and total S concentrations than wheat-soybean and wheat-sunflower rotation systems. Most test enzyme activities (&alpha;-galactosidase, &beta;-galactosidase, &alpha;-glucosidase, &beta;-glucosidase, urease, protease, phosphomonoesterase and arylsulphatase) showed the highest activities under wheat-corn intercropping system. Urease, protease and phosphodiesterase activities of wheat-cabbage sequential cropping system were significantly higher than two rotation systems. The maximum reaction rates of enzymes (V<sub>max</sub>) were higher than apparent enzyme activity, which suggests larger potential activity of enzymes, while not all kinetic parameters were adaptive as soil quality indicators in dry hilly cinnamon soil.

scholarly journals Subcellular distribution of glycosidases in human polymorphonuclear leucocytes

1978 ◽  
Vol 174 (1) ◽  
pp. 53-59 ◽  
Author(s):  
R F Rest ◽  
M H Cooney ◽  
J K Spitznagel
Keyword(s):  
Subcellular Distribution ◽  
Polymorphonuclear Leucocytes ◽  
Density Gradients ◽  
Glucosidase Activity ◽  
Mitochondrial Fractions ◽  
Specific Granules ◽  
Azurophil Granule ◽  
Alpha Galactosidase ◽  
Beta Glucosidase ◽  
Alpha Glucosidase

The subcellular distribution of nine glycosidases were studied in fractions of homogenized human polymorphonuclear leucocytes (neutrophils) obtained by isopycnic centrifugation through linear sucrose density gradients. The substrates were 4-methylumbelliferyl glycosides. All nine glycosides were hydrolysed by enzymes in neutrophil cytosol fractions, and by enzymes in at least one granule population. alpha-Glucosidase activity sedimented in sucrose density gradients to a point (p = 1.180 g/ml) just above the specific granules, possibly the ‘tertiary’ granule population. The peak corresponding to alpha-glucosidase did not co-sediment with, but considerably overlapped, the peak corresponding to lactoferrin, a marker for specific granules (p = 1.187 g/ml). alpha-Galactosidase activity was found primarily in heavy azurophil granules (p = 1.222 g/ml). alpha-Mannosidase activity was found primarily in light azurophil granules (p = 1.206 g/ml), following the distribution of myeloperoxidase, the commonly used azurophil granule marker. beta-Glucosidase activity was concentrated in mitochondrial fractions (p = 1.160 g/ml). All other glycosidases presented complex distributions, with activities not restricted to one granule class. Granule-associated glycosidase activities were increased 2–38 times when measured in the presence of 0.05% Triton X-100, indicating latency of the enzymes within granules.

scholarly journals Effects of a probiotic in combination with prebiotics on intestinal lactobacilli and coliforms and activities of bacterial enzymes in 1,2-dimethylhydrazine exposed rats

2011 ◽  
Vol 56 (No. 3) ◽  
pp. 99-106 ◽  
Author(s):  
L. Strojný ◽  
A. Bo ◽  
E. Hijová ◽  
A. Chmelárová ◽  
G. Mojžišová ◽  
...  
Keyword(s):  
High Fat Diet ◽  
Control Group ◽  
Group 4 ◽  
Group 3 ◽  
Group 5 ◽  
Group 2 ◽  
Beta Glucosidase ◽  
Alpha Glucosidase ◽  
Beta Galactosidase ◽  
Group 1

Effects of the probiotic (PRO) Lactobacillus plantarum and of the combination of PRO and the prebiotic (PRE) inulin enriched with oligofructose (2%), and PRO with Lini oleum virginale (O) on counts of lactobacilli and coliforms and enzymatic activities in faeces of rats were studied. The rats (n = 60) were divided into 5 groups of 12 subjects. The animals were fed on a high fat diet (10%) for 8 weeks of experiment. Colon cancer was induced by the application of 1,2-dimethylhydrazine (DMH) twice a week in a dose of 20 mg/kg s.c. in groups G2-G5. The rats in group 1 (control 1) received a diet without any supplements. The rats in group 2 (control 2) received 1,2 DMH without any supplements. The rats in group 3 received PRO, group 4 PRO and PRE, and group 5 received PRO and O. A significant decrease (P &lt; 0.05) of coliforms was found out after the application of PRO, PRO-O, and PRO-PRE in comparison with control group G2. Significantly higher (P &lt; 0.05) counts of lactobacilli were determined after the application of PRO-O and PRO-PRE. Significantly lower (P &lt; 0.001) activities of &beta;-galactosidase, &beta;-glucuronidase and &alpha;-glucosidase were observed in PRO, PRO-PRE and PRO-O, while in the case of the enzyme &beta;-glucosidase the activity was lower only after the addition of PRO-O. The protective effect of lactobacilli was observed in the order PRO-O, PRO-PRE, and PRO. It was shown that combinations of PRO-O and PRO-PRE had a synergistic effect which was higher than the effect of administering only PRO.

scholarly journals Specific inactivation of lysosomal glycosidases in living fibroblasts by the corresponding glycosylmethyl-p-nitrophenyltriazenes

1980 ◽  
Vol 188 (2) ◽  
pp. 337-343 ◽  
Author(s):  
O P Van Diggelen ◽  
H Galjaard ◽  
M L Sinnott ◽  
P J Smith
Keyword(s):  
Cell Viability ◽  
Enzyme Production ◽  
Zero Order ◽  
Fibroblast Cultures ◽  
Beta Glucosidase ◽  
Alpha Glucosidase ◽  
Beta Galactosidase ◽  
Lysosomal Glycosidases

1. beta-D-Galactopyranosylmethyl-p-nitrophenyltriazene can specificag detectable changes in cell viability or the activities of other hydrolases. 2. beta-D-Glucopyranosylmethyl-p-nitrophenyltriazene behaves similarly towards lysosomal beta-glucosidase, but also inactivates some alpha-glucosidase. 3. Both beta-galactosidase and beta-glucosidase activities in triazene-treated confluent fibroblast cultures recover exponentially; if zero-order enzyme production is assumed, turnover times of 10 and 5 days respectively can be estimated.

scholarly journals  α-Glucosidase and β-glucosidase from psychrotrophic strain arthrobacter sp. C2-2

2011 ◽  
Vol 23 (No. 3) ◽  
pp. 116-120 ◽  
Author(s):  
E. Benešová ◽  
M. Marková ◽  
B. Králová
Keyword(s):  
Exchange Chromatography ◽  
Gel Chromatography ◽  
Bacterial Strains ◽  
Molecular Weights ◽  
Ammonium Sulphate Precipitation ◽  
Donor And Acceptor ◽  
Alpha Galactosidase ◽  
Beta Glucosidase ◽  
Psychrotrophic Strain ◽  
Alpha Glucosidase

In this work six psychophilic and psychrotrophic bacterial strains were screened for the presence of different glycosidase activities (&alpha;-galactosidase, &alpha;-glucosidase, &beta;-glucosidase, &alpha;-mannosidase and &beta;-glucuronidase). Nine enzymes were found and their elementary characteristics were measured (t<sub>optimum</sub>, pH<sub>optimum</sub>, K<sub>m</sub>, V<sub>lim</sub>).Two enzymes with the highest activities at low temperatures were chosen for the next study, i.e. &alpha;-glucosidase and &beta;-glucosidase from the psychrotrophic strain Arthrobacter sp. C2-2. These enzymes were purified by ammonium sulphate precipitation, by chromatography with hydrophobic interaction, and by ion-exchange chromatography. Their molecular weights (&alpha;-glucosidase &ndash; 76 kDa, &beta;-glucosidase &ndash; 93 kDa) were determined by gel chromatography. In addition to this, it was verified that both of these enzymes are able to catalyse the transglycosylation reaction with the saccharidic donor and acceptor.

scholarly journals Decreased Activity of Blood Acid Sphingomyelinase in the Course of Multiple Myeloma

2019 ◽  
Vol 20 (23) ◽  
pp. 6048
Author(s):  
Marzena Wątek ◽  
Ewelina Piktel ◽  
Joanna Barankiewicz ◽  
Ewa Sierlecka ◽  
Sylwia Kościołek-Zgódka ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Human Blood ◽  
Blood Test ◽  
Acid Sphingomyelinase ◽  
Lysosomal Hydrolases ◽  
Healthy Group ◽  
Hematological Cancers ◽  
Beta Glucosidase ◽  
Beta Galactosidase

Acid sphingomyelinase (aSMase) is involved in the generation of metabolites that function as part of the sphingolipid signaling pathway. It catalyzes the breakdown of sphingomyelin into ceramide, a bioactive lipid that, among other roles, is involved in regulation of apoptosis. Dry drop blood test (DBS) and colorimetric 2-step enzymatic assay were used to assess the activity of human blood aSMase, beta-galactosidase, and beta-glucosidase, these enzymes are lysosomal hydrolases that catalyze the degradation of related sphingolipids, of sphingolipid signaling molecules. Blood was collected from a group of healthy volunteers and patients that were diagnosed with multiple myeloma (MM) in various stages of the disease. Additionally, activity of those enzymes in patients diagnosed with other hematological cancers was also assessed. We found that aSMase activity in the blood of patients with MM (at the time of diagnosis) was 305.43 pmol/spot*20 h, and this value was significantly lower (p < 0.030) compared to the healthy group 441.88 pmol/spot*20 h. Our collected data suggest a possible role of aSMase in pathogenesis of MM development.

scholarly journals Acid hydrolases in leukocytes and platelets of normal subjects and in patients with Gaucher's and Fabry's disease.

1976 ◽  
Vol 143 (4) ◽  
pp. 975-980 ◽  
Author(s):  
E Beutler ◽  
W Kuhl ◽  
F Matsumoto ◽  
G Pangalis
Keyword(s):  
Acid Phosphatase ◽  
Normal Subjects ◽  
Acid Hydrolases ◽  
Gaucher's Disease ◽  
Gaucher’S Disease ◽  
Fabry’S Disease ◽  
Fabry's Disease ◽  
Alpha Galactosidase ◽  
Beta Glucosidase ◽  
Alpha Glucosidase

Lymphocytes, monocytes, neutrophilic granulocytes and platelets were each separated to greater than 95% purity from six normal subjects, three patients with Gaucher's disease, two heterozygotes for Gaucher's disease, and one patient with Fabry's disease. Activities of the following acid hydrolases were determined: "acid" (pH 4.0) beta-glucosidase, pH 5.0 beta-glucosidase, alpha-galactosidase, alpha-arabinosidase, alpha-mannosidase, alpha-glucosidase, beta-glucuronidase, beta-galactosidase, beta-hexosaminidase, and acid phosphatase. Enzymatic activity varied greatly with cell type and the enzyme being measured; the importance of assaying pure preparations especially for heterozygote detection is emphasized. Gaucher's disease patients' cells were found to be deficient in the pH 4.0 acid beta-glucosidase, variable in the pH 5.0 beta-glucosidase, and normal in all other acid hydrolases tested, including acid phosphatase, the activity of which is known to be elevated in plasma. Blood cells of a patient with Fabry's disease were deficient in alpha-galactosidase and normal in all other acid hydrolases tested.

scholarly journals Purification and properties of a stable β-glucosidase from an extremely thermophilic anaerobic bacterium

1987 ◽  
Vol 243 (3) ◽  
pp. 779-787 ◽  
Author(s):  
M L Patchett ◽  
R M Daniel ◽  
H W Morgan
Keyword(s):  
Anaerobic Bacterium ◽  
Thermal Inactivation ◽  
First Order ◽  
First Order Kinetics ◽  
Purification And Properties ◽  
Wide Range ◽  
Optimum Ph ◽  
Exclusion Chromatography ◽  
Beta Glucosidase ◽  
Beta Galactosidase

A beta-glucosidase (EC 3.2.1.21) was purified to homogeneity from cell-free extracts of an extremely thermophilic anaerobic bacterium. The enzyme has an Mr of 43,000 as determined by molecular-exclusion chromatography, has a pI of 4.55 and shows optimum activity at pH 6.2. The enzyme is active against a wide range of aryl beta-glycosides and beta-linked disaccharides, with beta-galactosidase activity only slightly less than beta-glucosidase activity, and significant beta-xylosidase activity. Lineweaver-Burk plots for p-nitrophenyl beta-glucoside, o-nitrophenyl beta-glucoside and cellobiose substrates are biphasic concave-downwards. Inhibition of the beta-glucosidase by substrates and glucose is negligible. Thermal inactivation follows first-order kinetics, with t1/2 (65 degrees C) 45 h, t1/2 (75 degrees C) 47 min and t1/2 (85 degrees C) 1.4 min and a deactivation energy of 380 kJ/mol at pH 6.2. At pH 7.0, which is the optimum pH for thermostability, t1/2 (75 degrees C) is 130 min. At 75 degrees C, at pH 6.2, the thermostability is enhanced about 8-fold by 10% (w/v) glycerol, about 6-fold by 0.2 M-cellobiose and about 3-fold by 5 mM-dithiothreitol and 5 mM-2-mercaptoethanol.

scholarly journals Absence of step changes in activity of certain enzymes during the cell cycle of budding and fission yeasts in synchronous cultures

1983 ◽  
Vol 61 (1) ◽  
pp. 339-349
Author(s):  
J. Creanor ◽  
S.G. Elliott ◽  
Y.C. Bisset ◽  
J.M. Mitchison
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Acid Phosphatase ◽  
Kluyveromyces Lactis ◽  
The Other ◽  
Linear Pattern ◽  
Synchronous Cultures ◽  
Asynchronous Control ◽  
Alpha Glucosidase ◽  
Beta Galactosidase

Synchronous cultures prepared by selection from an elutriating rotor were used to measure activity changes during the cell cycle of the following enzymes: acid phosphatase in Schizosaccharomyces pombe and Saccharomyces cerevisiae, alpha-glucosidase in S. cerevisiae and beta-galactosidase in Kluyveromyces lactis. There was no sign of step rises in activity in acid phosphatase but there were indications in S. cerevisiae of the linear pattern with rate doublings once per cycle that had been found previously in S. pombe. There was also no sign of step rises in the other two enzymes, in contrast to earlier results using different techniques. Asynchronous control cultures showed little or no perturbations after the first hour.

scholarly journals Purification and properties of a β-1,6-glucanase from Penicillium brefeldianum

1984 ◽  
Vol 223 (3) ◽  
pp. 707-714 ◽  
Author(s):  
G P Schep ◽  
M G Shepherd ◽  
P A Sullivan
Keyword(s):  
High Pressure ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Dry Weight ◽  
Protein Band ◽  
Purification And Properties ◽  
Freeze Dried ◽  
Beta Glucosidase

An inducible endo-beta-1,6-glucanase was purified from Penicillium brefeldianum by DEAE-cellulose, Bio-Gel P-150 and high-pressure liquid chromatography. The final preparation was essentially free from beta-1,3-glucanase and beta-glucosidase activities. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis revealed one protein band with an Mr of 44000. The Vmax. and Km values were calculated to be 624 units (mumol/min)/mg and 2.78 mg/ml respectively. The glucanase had lytic activity against mycelial cells of the yeast Candida albicans. The yield of purified beta-1,6-glucanase from 100 mg dry weight of freeze-dried culture filtrate varied from 60 to 180 units.

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