scholarly journals Mechanism of the reaction catalysed by cytosolic 5′-nucleotidase/phosphotransferase: formation of a phosphorylated intermediate

1996 ◽  
Vol 317 (3) ◽  
pp. 797-801 ◽  
Author(s):  
Cristina BAIOCCHI ◽  
Rossana PESI ◽  
Marcella CAMICI ◽  
Roichi ITOH ◽  
Maria GRAZIA TOZZI
Keyword(s):  
Reaction Mechanism ◽  
Kinetic Parameters ◽  
Acid Treatment ◽  
Histidine Residue ◽  
Thiol Oxidation ◽  
Photo Oxidation ◽  
Calf Thymus ◽  
Hydrolysis Of ◽  
Mechanism Of The Reaction

Cytosolic 5´-nucleotidase preferentially catalysing the hydrolysis of IMP, GMP and their deoxy derivatives, and endowed with phosphotransferase activity, was purified from calf thymus and its reaction mechanism was studied. In the presence of [32P]IMP, ATP and MgCl2, a covalent enzyme–phosphate intermediate was trapped by mixing with an SDS solution. Heat or acid treatment of the enzyme before incubation with radiolabelled substrate prevented formation of the intermediate. Furthermore, on the basis of studies on the kinetic parameters of the enzyme as function of pH, and of experiments on thiol oxidation and photo-oxidation, we suggest the involvement of cysteine and histidine residue(s) in the reaction mechanism.

Chemical mechanism of β-glucosidase from Trichoderma reesei QM 9414. pH-dependence of kinetic parameters

1992 ◽  
Vol 283 (3) ◽  
pp. 679-682 ◽  
Author(s):  
I de la Mata ◽  
P Estrada ◽  
R Macarrón ◽  
J M Dominguez ◽  
M P Castillón ◽  
...  
Keyword(s):  
Kinetic Parameters ◽  
Trichoderma Reesei ◽  
Carboxy Group ◽  
Ph Dependence ◽  
Competitive Inhibitor ◽  
Chemical Mechanism ◽  
Histidine Residue ◽  
Activity Temperature ◽  
Beta Glucosidase ◽  
Mechanism Of The Reaction

The variation of kinetic parameters of beta-glucosidase from Trichoderma reesei QM 9414 with pH was used to gain information about the chemical mechanism of the reaction catalysed by this enzyme. The pH-dependence of Vmax. and Vmax./Km for p-nitrophenyl beta-D-glucopyranoside showed that a group with a pK value of 4.3 must be unprotonated and a group with a pK value of 5.9 must be protonated for activity. Temperature and solvent-perturbation studies indicated that these groups are a histidine residue and a carboxy group respectively. Profiles of pKi for maltose as competitive inhibitor showed that binding is prevented when a group on the enzyme with a pK value of 4.5 becomes protonated.

scholarly journals Substrate specificity and kinetic properties of α-galactosidases from Vicia faba

1969 ◽  
Vol 115 (1) ◽  
pp. 47-54 ◽  
Author(s):  
P. M. Dey ◽  
J. B. Pridham
Keyword(s):  
Catalytic Activity ◽  
Substrate Specificity ◽  
Kinetic Parameters ◽  
Vicia Faba ◽  
Active Site ◽  
Effect Of Temperature ◽  
Kinetic Properties ◽  
Photo Oxidation ◽  
Hydrolysis Of ◽  
Enzyme I

1. The hydrolysis of a variety of galactosides and other glycosides by α-galactosidases I and II of Vicia faba was studied. 2. The effect of temperature on kinetic parameters was also examined. 3. Both enzymes are inhibited by excess of substrate (p-nitrophenyl α-d-galactoside); with enzyme I this is competitive and is caused by the galactosyl moiety. 4. Enzyme I is inhibited by oligosaccharides possessing terminal non-reducing galactose residues and to a smaller extent by l-arabinose and d-fucose. 5. The effect of pH on Km and Vmax. values suggests that carboxyl and imidazole groups are involved in the catalytic activity of enzyme I. 6. Photo-oxidation experiments with enzyme I also suggest that an imidazole group is present at the active site.

Influence of medium on alkaline hydrolysis of succinic acid monomethyl and monopropyl esters

1980 ◽  
Vol 45 (11) ◽  
pp. 2873-2882
Author(s):  
Vladislav Holba ◽  
Ján Benko
Keyword(s):  
Succinic Acid ◽  
Kinetic Parameters ◽  
Alkaline Hydrolysis ◽  
Specific Interactions ◽  
Nonaqueous Media ◽  
The Media ◽  
Kinetics Of ◽  
Hydrolysis Of

The kinetics of alkaline hydrolysis of succinic acid monomethyl and monopropyl esters were studied in mixed aqueous-nonaqueous media at various temperatures and ionic strengths. The results of measurements are discussed in terms of electrostatic and specific interactions between the reactants and other components of the reaction mixture. The kinetic parameters in the media under study are related to the influence of the cosolvent on the solvation sphere of the reactants.

Vinyl ether hydrolysis. XVII. Oxacyclonon-2,8-diene and the question of the unorthodox reaction mechanism for 9-methoxyoxacyclonon-2-ene

1984 ◽  
Vol 62 (1) ◽  
pp. 74-76 ◽  
Author(s):  
R. A. Burt ◽  
Y. Chiang ◽  
A. J. Kresge ◽  
S. Szilagyi
Keyword(s):  
Reaction Mechanism ◽  
Vinyl Ether ◽  
Membered Ring ◽  
Specific Rate ◽  
Ether Group ◽  
Reaction Proceeds ◽  
Great Reactivity ◽  
Rate Of Hydrolysis ◽  
Acid Catalyzed ◽  
Hydrolysis Of

The acid-catalyzed hydrolysis of the nine-membered ring cyclic vinyl ether, oxacyclonon-2,8-diene, occurs with a normal isotope effect, [Formula: see text], which indicates that this reaction proceeds by the conventional vinyl ether hydrolysis mechanism involving rate-determining proton transfer to carbon. The specific rate of this reaction, [Formula: see text], may then be used to show that there is no significant ring-size effect on the rate of hydrolysis of a vinyl ether group in a nine-membered ring. The previously noted unusually great reactivity of the vinyl ether group in 9-methoxyoxacyclonon-2-ene, for which an unorthodox reaction mechanism has been claimed, must therefore be due to some other cause.

scholarly journals Optimized acid hydrolysis of the polysaccharides from the seaweed Solieria filiformis (Kützing) P.W. Gabrielson for bioethanol production

2017 ◽  
Vol 39 (4) ◽  
pp. 423 ◽  
Author(s):  
George Meredite Cunha de Castro ◽  
Norma Maria Barros Benevides ◽  
Maulori Curié Cabral ◽  
Rafael De Souza Miranda ◽  
Enéas Gomes Filho ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Sulfuric Acid ◽  
Acid Hydrolysis ◽  
Kinetic Parameters ◽  
Renewable Resource ◽  
Bioethanol Production ◽  
Seaweed Species ◽  
Mild Conditions ◽  
Hydrolysis Of ◽  
Solieria Filiformis

 The seaweeds are bio-resource rich in sulfated and neutral polysaccharides. The tropical seaweed species used in this study (Solieria filiformis), after dried, shows 65.8% (w/w) carbohydrate, 9.6% (w/w) protein, 1.7% (w/w) lipid, 7.0% (w/w) moisture and 15.9% (w/w) ash. The dried seaweed was easily hydrolyzed under mild conditions (0.5 M sulfuric acid, 20 min.), generating fermentable monosaccharides with a maximum hydrolysis efficiency of 63.21%. Galactose and glucose present in the hydrolyzed were simultaneously fermented by Saccharomyces cerevisiae when the yeast was acclimated to galactose and cultivated in broth containing only galactose. The kinetic parameters of the fermentation of the seaweed hydrolyzed were Y(P⁄S) = 0.48 ± 0.02 g.g−1, PP = 0.27 ± 0.04 g.L−1.h−1, h = 94.1%, representing a 41% increase in bioethanol productivity. Therefore, S. filiformis was a promising renewable resource of polysaccharides easily hydrolyzed, generating a broth rich in fermentable monosaccharides for ethanol production. 

The crystal structure of human mitochondrial 3-ketoacyl-CoA thiolase (T1): insight into the reaction mechanism of its thiolase and thioesterase activities

2014 ◽  
Vol 70 (12) ◽  
pp. 3212-3225 ◽  
Author(s):  
Tiila-Riikka Kiema ◽  
Rajesh K. Harijan ◽  
Malgorzata Strozyk ◽  
Toshiyuki Fukao ◽  
Stefan E. H. Alexson ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Reaction Mechanism ◽  
Active Site ◽  
Quaternary Structure ◽  
Fatty Acyl ◽  
Physiological Importance ◽  
Loop Domain ◽  
Solution Studies ◽  
Hydrolysis Of ◽  
Insight Into

Crystal structures of human mitochondrial 3-ketoacyl-CoA thiolase (hT1) in the apo form and in complex with CoA have been determined at 2.0 Å resolution. The structures confirm the tetrameric quaternary structure of this degradative thiolase. The active site is surprisingly similar to the active site of theZoogloea ramigerabiosynthetic tetrameric thiolase (PDB entries 1dm3 and 1m1o) and different from the active site of the peroxisomal dimeric degradative thiolase (PDB entries 1afw and 2iik). A cavity analysis suggests a mode of binding for the fatty-acyl tail in a tunnel lined by the Nβ2–Nα2 loop of the adjacent subunit and the Lα1 helix of the loop domain. Soaking of the apo hT1 crystals with octanoyl-CoA resulted in a crystal structure in complex with CoA owing to the intrinsic acyl-CoA thioesterase activity of hT1. Solution studies confirm that hT1 has low acyl-CoA thioesterase activity for fatty acyl-CoA substrates. The fastest rate is observed for the hydrolysis of butyryl-CoA. It is also shown that T1 has significant biosynthetic thiolase activity, which is predicted to be of physiological importance.

Theoretical study on the reaction mechanism for the hydrolysis of esters and amides under acidic conditions

Tetrahedron ◽  
2007 ◽  
Vol 63 (5) ◽  
pp. 1264-1269 ◽  
Author(s):  
Kenzi Hori ◽  
Yutaka Ikenaga ◽  
Kouichi Arata ◽  
Takanori Takahashi ◽  
Kenji Kasai ◽  
...  
Keyword(s):  
Reaction Mechanism ◽  
Theoretical Study ◽  
Acidic Conditions ◽  
Hydrolysis Of Esters ◽  
Hydrolysis Of

scholarly journals Kinetics and Reaction Mechanism for Alkaline Hydrolysis of Y-Substituted-Phenyl Diphenylphosphinates

2013 ◽  
Vol 34 (7) ◽  
pp. 2001-2005 ◽  
Author(s):  
Hyo-Jeong Hong ◽  
Jieun Lee ◽  
Ae Ri Bae ◽  
Ik-Hwan Um
Keyword(s):  
Reaction Mechanism ◽  
Alkaline Hydrolysis ◽  
Hydrolysis Of
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