Hydrolysis of steroid glucuronides with beta-glucuronidase preparations from bovine liver, Helix pomatia, and E. coli.

1977 ◽  
Vol 23 (3) ◽  
pp. 532-535 ◽  
Author(s):  
V Graef ◽  
E Furuya ◽  
O Nishikaze
Keyword(s):  
Helix Pomatia ◽  
Bovine Liver ◽  
Enzymic Activity ◽  
Enzyme Preparations ◽  
E Coli ◽  
Rate Of Hydrolysis ◽  
Polystyrene Resin ◽  
Good For ◽  
Hydrolysis Of ◽  
Urinary Steroid

Abstract We determined the enzymic activity of beta-glucuronidase preparations from bovine liver, Helix pomatia, and Escherischia coli with steroid glucuronides and nonsteroid glucuronides as substrates. We also studied the effect of Na2SO4 on the enzymic hydrolysis of several substrates with the three preparations of beta-glucuronidase. Na2SO4 increases the rate of hydrolysis of all substrates with beta-glucuronidase from bovine liver. Hydrolysis of a steroid glucoronide with beta-glucuronidase from Helix pomatia and E. coli is inhibited by Na2SO4. None of the three enzyme preparations gives complete hydrolysis of urinary steroid conjugates, because urine contains inhibitors, which can be removed by absorption chromatography of the urine on a column of neutral polystyrene resin Amberlite XAD-2. But when Amberlite XAD-2 is not used, hydrolysis of urinary glucuronides of androsterone, etiocholanolone, pregnanediol, estriol, and 17-hydroxycorticosteroids proves that, given an incubation time of 24 h, the beta-glucuronidase preparation from bovine liver, in the presence of Na2SO4, is suited for determining all of the above steroids except esriol; the preparation from Helix pomatia is good for determining estriol and 17-hydroxycorticosteroids; the preparation from E. coli is good for determining androsterone, 17-hydroxycorticosteroids, and especially estriol, the glucuronide, of which is maximally hydrolyzed in 2 h.

scholarly journals The hydrolysis of glycosyl fluorides by glycosidases

1967 ◽  
Vol 105 (2) ◽  
pp. 669-672 ◽  
Author(s):  
J. E. G. Barnett ◽  
W. T. S. Jarvis ◽  
K A Munday
Keyword(s):  
Helix Pomatia ◽  
Enzyme Preparations ◽  
Intestinal Juice ◽  
Hydrolysis Of ◽  
Specific Inhibitors

1. α-d- and β-d-Glucopyranosyl, α-d- and β-d-galactopyranosyl, α-d-mannopyranosyl and α-d-xylopyranosyl fluorides were hydrolysed specifically by the respective glycosidases from several sources. 2. Use of specific inhibitors with a mixture of glycosidases from Helix pomatia intestinal juice showed that each glycosyl fluoride was hydrolysed only by the respective glycosidase. α-d-Glucopyranosidase and α-d-xylopyranosidase activities were shown to be due to different enzymes. 3. Partially purified enzyme preparations containing only one of the glycosidase activities hydrolysed only the corresponding glycosyl fluoride. 4. The configuration at C-1 of α-d-mannopyranosyl fluoride was confirmed since it was hydrolysed by an α-d-mannosidase preparation that contained no detectable β-d-mannosidase activity. 5. An attempt to prepare o-nitrophenyl β-d-mannopyranoside led only to o-nitrophenyl α-d-mannopyranoside.

Stability and Hydrolysis of Desomorphine-Glucuronide

2019 ◽  
Vol 43 (7) ◽  
pp. 536-542 ◽  
Author(s):  
Jessica Winborn ◽  
Sarah Kerrigan
Keyword(s):  
Glucuronic Acid ◽  
Helix Pomatia ◽  
Sample Treatment ◽  
E Coli ◽  
Hydrolysis Method ◽  
Flight Mass Spectrometry ◽  
The Stability ◽  
Hydrolysis Of ◽  
Additional Sample ◽  
Analgesic Potency

AbstractDesomorphine, the principal opioid in Krokodil, has an analgesic potency approximately ten-times that of morphine. Similar to other opioids, during phase II metabolism it undergoes conjugation with glucuronic acid to form desomorphine-glucuronide. Although hydrolysis of conjugated species is sometimes required prior to analysis, desomorphine-glucuronide has not been fully investigated. In this study, six hydrolysis procedures were optimized and evaluated. Deconjugation efficiencies using chemical and enzymatic hydrolysis were evaluated and stability in aqueous solution was assessed. Acid hydrolysis was compared with five β-glucuronidase sources (BGTurbo™, IMCSzyme™, Escherichia coli, Helix pomatia and Patella vulgata). At optimal conditions, each hydrolysis method produced complete hydrolysis (≥96%). However, under simulated challenging conditions, P. vulgata was the most efficient β-glucuronidase for the hydrolysis of desomorphine-glucuronide. Both BGTurbo™ and IMCSzyme™ offered fast hydrolysis with no need for sample cleanup prior to liquid chromatography-quadrupole/time of flight-mass spectrometry (LC-Q/TOF-MS) analysis. Hydrolysates using E. coli, H. pomatia and P. vulgata underwent additional sample treatment using β-Gone™ cartridges. Additionally, the stability of free and conjugated drug was evaluated at elevated temperature (60°C) in aqueous solutions between pH 4 and 10. No degradation was observed for either desomorphine or desomorphine-glucuronide under any of the conditions tested.

scholarly journals The hydrolysis of phosphatidylinositol monolayers at an air/water interface by the calcium-ion-dependent phosphatidylinositol phosphodiesterase of pig brain

1981 ◽  
Vol 193 (2) ◽  
pp. 607-614 ◽  
Author(s):  
K Hirasawa ◽  
R F Irvine ◽  
R M C Dawson
Keyword(s):  
Oleyl Alcohol ◽  
Enzymic Activity ◽  
Lag Phase ◽  
Water Interface ◽  
Inhibitory Effects ◽  
Pig Brain ◽  
Air Water Interface ◽  
Rate Of Hydrolysis ◽  
Hydrolysis Of

1. The activity of Ca2+-dependent phosphatidylinositol phosphodiesterase (EC 3.1.4.10) of pig brain against [32P]phosphatidylinositol monolayers at an air/water interface has been measured. As the monolayer pressure was increased a sharp cut-off of enzymic hydrolysis occurred at 33 × 10(-3) N/m. 2. The addition of either phosphatidic acid, phosphatidylglycerol or oleyl alcohol increased the film pressure at which cut off occurred, as well as increasing the rate of hydrolysis at lower pressures. 3. The rate of hydrolysis, but not the cut-off pressure, was markedly increased by oleic acid and slightly increased by phosphatidylethanolamine. 4. Phosphatidylcholine, palmitoylcholine and octadecylamine decreased the cut-off pressure, as well as the enzymic activity below this pressure. 5. Stearic acid and stearyl alcohol had no effect on either the cut-off pressure or the activity. 6. All activators decreased the length of the lag phase before enzyme activity began, and phosphatidylcholine increased it. 7. These results are compared with the stimulatory and inhibitory effects of various amphiphiles observed previously with phosphatidylinositol dispersions [Irvine, Hemington & Dawson (1979) Eur. J. Biochem. 99, 525-530], and their possible relevance to the control of the phosphatidylinositol phosphodiesterase in vivo are discussed.

scholarly journals Uji Aktivitas Sitotoksik dan Antibakteri Ekstrak Daun Tumbuhan Rengas (Gluta renghas L)

2020 ◽  
Vol 11 (1) ◽  
pp. 52-60
Author(s):  
Sanusi Ibrahim ◽  
Suryati Suryati ◽  
Enda Desriansyah Aziz
Keyword(s):  
Antibacterial Activity ◽  
Ethyl Acetate ◽  
Methanol Extract ◽  
Ethyl Acetate Extract ◽  
Inhibition Zone ◽  
Cytotoxicity Activity ◽  
E Coli ◽  
Rate Of Hydrolysis ◽  
Hydrolysis Of ◽  
Ethyl Acetate Extracts

Generally, Gluta renghas L. is known because of its very toxic latex which can cause hard irritations to skin. Although, rengas latex has efficacy as an antibacterial agent. Related to previous research, it has been reported that there are urisol, rengol, glutarengol, laccol, and thitsiol in rengas latex. The woody trunk was reported to contain flavonoid, benzenoid, lipid, and steroid compounds. The leaves, were reported shows that the leaves of G. renghas contain an anticholinesterase substance and can be reduced the rate of hydrolysis of acetylcholine. Extraction of rengas leaves has been carried out. It has been shown phenols, steroids, and coumarins compounds in methanol extract, flavonoids, phenols, saponins and alkaloids compounds in ethyl acetate extract, steroids and alkaloids compounds in hexane extract. In the present, cytotoxicity and antibacterial activity have been tested. The results show that the great cytotoxicity activity by ethyl acetate extracts which have an LC50 value is 123,718 µg/mL (R2 0.9822), while the great antibacterial activity shown by methanol extract 1,000 µg/mL with a diameter of inhibition zone 19.02 mm (S. aureus) and 16.06 mm (E. coli). 

A ribosomal-boand aminopeptidase in Escherichia coli B: substrate specificity

1970 ◽  
Vol 48 (12) ◽  
pp. 1292-1296 ◽  
Author(s):  
A. T. Matheson ◽  
A. J. Dick ◽  
F. Rollin
Keyword(s):  
Escherichia Coli ◽  
Substrate Specificity ◽  
Peptide Bond ◽  
Terminal Amino Acid ◽  
Strong Activity ◽  
E Coli ◽  
Rate Of Hydrolysis ◽  
Terminal Amino ◽  
Hydrolysis Of ◽  
Escherichia Coli B

The substrate specificity of the ribosomal-bound aminopeptidase from Escherichia coli B has been studied using di-, tri-, and tetrapeptides. The enzyme shows strong activity to leucyl, methionyl, threonyl, and lysyl peptides. Of the other dipeptides tested considerable hydrolysis was observed only if the C-terminal amino acid was leucine or methionine. In a given series of peptides the rate of hydrolysis of the N-terminal peptide bond increased as the size of the peptide increased. Although leucyi dipeptides were hydroiyzed more rapidly than the corresponding methionyl dipeptide the reverse was true with the tripeptides tested. No carboxypeptidase activity was observed and peptides containing D-amino acids were not hydroiyzed. The substrate specificity of the aminopeptidase was compared with the known N-terminal sequences of E. coli proteins to determine whether the enzyme may be involved in the removal of N-formylmethionyl from newly synthesized polypeptides.

scholarly journals STUDIES ON THE LACTASE OF ESCHERICHIA COLI

1941 ◽  
Vol 24 (3) ◽  
pp. 377-397 ◽  
Author(s):  
H. P. Knopfmacher ◽  
A. J. Salle
Keyword(s):  
Escherichia Coli ◽  
Sodium Sulfide ◽  
Potassium Cyanide ◽  
Order Reaction ◽  
E Coli ◽  
First Order ◽  
Optimal Activity ◽  
Rate Of Hydrolysis ◽  
Hydrolysis Of ◽  
Slight Inhibition

A "lactase solution" was prepared from Escherichia coli. The mechanism of its action has been studied and changes in the rate of hydrolysis under various conditions investigated. The hydrolysis of lactose by the enzyme approximates the course of reaction of the integrated Michaelis-Menten equation. One molecule of enzyme combines with one molecule of substrate. E. coli lactase is readily inactivated at pH 5.0, and its optimal activity at 36°C. is reached between pH 7.0 and pH 7.5. The optimal temperature for its action was found to be 46°C. when determinations were carried out after an incubation period of 30 minutes. Its inactivation by heat follows the course of a first order reaction, and the critical thermal increment between the temperatures of 45°C. and 53°C. was calculated to be 56,400 calories per mol. The enzyme is activated by potassium cyanide, sodium sulfide, and cysteine, and irreversibly inactivated by mercuric chloride, silver nitrate, and iodine. After inactivation with copper sulfate partial reactivation is possible, while the slight inhibition brought about by hydrogen peroxide is completely reversible. The possible structure of the active groups of E. coli lactase as compared with other enzymes has been discussed.

Morphine glucuronide hydrolysis: superiority of beta-glucuronidase from Patella vulgata.

1982 ◽  
Vol 28 (1) ◽  
pp. 83-86 ◽  
Author(s):  
J Combie ◽  
J W Blake ◽  
T E Nugent ◽  
T Tobin
Keyword(s):  
Reaction Rate ◽  
Helix Pomatia ◽  
Cost Effective ◽  
Bovine Liver ◽  
Enzyme Concentration ◽  
Urine Samples ◽  
Factors Affecting ◽  
Patella Vulgata ◽  
Hydrolysis Of ◽  
Morphine Glucuronide

Abstract beta-Glucuronidase from Patella vulgata, Helix aspersa, Helix pomatia, and bovine liver were evaluated for usefulness in routine hydrolysis of drug-glucuronic acid conjugates from equine urine samples. Factors affecting the reaction rate (enzyme concentration, ligand concentration, temperature, and pH) were optimized. A 3-h incubation at 65 degrees C with 5000 U of beta-glucuronidase from P. vulgata per milliliter of urine resulted in complete hydrolysis of all morphine glucuronide in the urine samples. Not only was the enzyme preparation from P. vulgata the most cost-effective beta-glucuronidase source studied, but also its thermal stability is such that it can be used at a temperature high enough to substantially shorten the incubation interval. Preliminary work on other drugs that form glucuronide conjugates indicates that this same procedure is similarly superior for use in their hydrolysis.

scholarly journals Identification of bovine liver mitochondrial NAD+ glycohydrolase as ADP-ribosyl cyclase

1997 ◽  
Vol 326 (2) ◽  
pp. 401-405 ◽  
Author(s):  
Mathias ZIEGLER ◽  
Dierk JORCKE ◽  
Manfred SCHWEIGER
Keyword(s):  
Calcium Release ◽  
Bovine Liver ◽  
Enzymic Activity ◽  
Cyclase Activity ◽  
Partial Recovery ◽  
Nad Glycohydrolase ◽  
Adp Ribosyl Cyclase ◽  
Cyclic Adp Ribose ◽  
Hydrolysis Of ◽  
Synthesis And Degradation

The present investigation identifies bovine liver mitochondrial NADase (NAD+ glycohydrolase) as a member of the class of bifunctional ADP-ribosyl cyclases/cyclic ADP-ribose hydrolases, known to be potential second messenger enzymes. These enzymes catalyse the synthesis and degradation of cyclic ADP-ribose, a potent intracellular calcium-mobilizing agent. The mitochondrial enzyme utilized the NAD+ analogues nicotinamide guanine dinucleotide (NGD+) and nicotinamide hypoxanthine dinucleotide (NHD+) to form fluorescent cyclic purine nucleoside diphosphoriboses. ADP-ribosyl cyclase activity was also demonstrated using 32P-labelled NAD+ as substrate. The identity of NADase and ADP-ribosyl cyclase was supported by their co-migration in SDS/polyacrylamide gels. Cyclase activity was visualized directly within the gel by detecting the formation of fluorescent cyclic IDP-ribose from NHD+. The enzyme catalysed the hydrolysis of cyclic ADP-ribose to ADP-ribose. Moreover, in the presence of nicotinamide and cyclic ADP-ribose the enzyme synthesized NAD+. Both the ADP-ribosyl cyclase and NADase activities of the enzyme were strongly inhibited by reducing agents. Treatment of the NADase with dithiothreitol caused the apparent inactivation of the enzyme. Subsequent removal of the reducing agent and addition of oxidized glutathione led to a partial recovery of enzymic activity. The results support a model for pro-oxidant-induced calcium release from mitochondria involving cyclic ADP-ribose as a specific messenger, rather than the non-enzymic modification of proteins by ADP-ribose.

Improved hydrolysis of urinary 17-hydroxycorticosteroid glucuronides with beta-glucuronidase from Helix pomatia, on adding sodium sulfate.

1977 ◽  
Vol 23 (12) ◽  
pp. 2332-2334 ◽  
Author(s):  
O Nishikaze ◽  
T Kobayashi
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Inhibitory Activity ◽  
Sodium Sulfate ◽  
Helix Pomatia ◽  
Bovine Liver ◽  
Final Concentration ◽  
Urine Samples ◽  
Analytical Recovery ◽  
Hydrolysis Of

Abstract Sodium sulfate increases the hydrolysis of urinary 17-hydroxycorticosteroid glucuronides with beta-glucuronidase preparations derived from Helix pomatia because it removes the inhibitory activity of urinary high-molecular-weight substances. For maximum hydrolysis of urinary 17-hydroxycorticosteroid glucuronides, the hydrolysis [5 ml of urine, 0.5 ml of 2 mol/liter acetate buffer (pH 5.0)] should be conducted in the presence of sodium sulfate (final concentration: 80 g/liter) with (a) 600 Fishman units of the enzyme per milliliter of urine (18 h at 52 degrees C) or (b) with 1500 units of the enzyme per milliliter of urine (3 h at 57 degrees C). Under conditions a, analytical recovery of steroid glucuronides added to 12 urine samples was 99 +/- 2.1% (96-102%). Values obtained for 20 urine samples with this method were 99 +/- 2.7% (93-104%) as great as those yielded by a method in which 600 units of the enzyme from bovine liver are used together with sodium sulfate (18 h at 48 degrees C).

Sign in / Sign up

Export Citation Format

Share Document