scholarly journals The kinetics of hydrolysis of some synthetic substrates containing neutral hydrophilic groups by pig pepsin and chicken liver cathepsin D.

1983 ◽  
Vol 211 (1) ◽  
pp. 237-242 ◽  
Author(s):  
G B Irvine ◽  
N L Blumsom ◽  
D T Elmore
Keyword(s):  
Cathepsin D ◽  
Order Rate Constant ◽  
Chicken Liver ◽  
Lag Phase ◽  
Hydrophilic Group ◽  
Steady State Kinetics ◽  
Cationic Group ◽  
Specificity Constant ◽  
Kinetics Of ◽  
Hydrolysis Of

1. Several peptides containing either of the sequences -Phe(NO2)-Trp- and -Phe(NO2)-Phe- and an uncharged hydrophilic group were synthesized, and the steady-state kinetics of their hydrolysis by pig pepsin (EC 3.4.23.1) and chicken liver cathepsin D (EC 3.4.23.5) were determined. Despite the presence of a hydrophilic group to increase substrate solubility, it was not possible to achieve the condition [S]0 much greater than Km, and, in some cases, only values of kcat./Km could be determined by measuring the first-order rate constant when [S]0 much less than Km. 2. Occupancy of the P2 and P3 sites considerably enhanced the specificity constant, and alanine was more effective than glycine at site P2. 3. The specificity constants for the hydrolysis by pepsin of those substrates in the present series that contain an amino acid residue at site P3 are considerably lower than for comparable substrates containing a cationic group. This difference does not apply to cathepsin D. 4. Hydrolyses with cathepsin D commonly exhibited a lag phase, and a possible explanation for this is given.

Steady-state and pre-steady-state kinetics of the mitochondrial F(1)F(o) ATPase: is ATP synthase a reversible molecular machine?

2000 ◽  
Vol 203 (1) ◽  
pp. 41-49 ◽  
Author(s):  
A.D. Vinogradov
Keyword(s):  
Steady State ◽  
Atp Synthase ◽  
Atp Synthesis ◽  
Gamma Subunit ◽  
Change Mechanism ◽  
Steady State Kinetics ◽  
Energy Dependent ◽  
Variable Substrate ◽  
Kinetics Of ◽  
Hydrolysis Of

H(+)-ATP synthase (F(1)F(o) ATPase) catalyzes the synthesis and/or hydrolysis of ATP, and the reactions are strongly affected by all the substrates (products) in a way clearly distinct from that expected of a simple reversibly operating enzyme. Recent studies have revealed the structure of F(1), which is ideally suited for the alternating binding change mechanism, with a rotating gamma-subunit as the energy-driven coupling device. According to this mechanism ATP, ADP, inorganic phosphate (P(i)) and Mg(2+) participate in the forward and reverse overall reactions exclusively as the substrates and products. However, both F(1) and F(1)F(o) demonstrate non-trivial steady-state and pre-steady-state kinetics as a function of variable substrate (product) concentrations. Several effectors cause unidirectional inhibition or activation of the enzyme. When considered separately, the unidirectional effects of ADP, P(i), Mg(2+) and energy supply on ATP synthesis or hydrolysis may possibly be explained by very complex kinetic schemes; taken together, the results suggest that different conformational states of the enzyme operate in the ATP hydrolase and ATP synthase reactions. A possible mechanism for an energy-dependent switch between the two states of F(1)F(o) ATPase is proposed.

scholarly journals Kinetics of the alkaline hydrolysis of crystal violet in micelles, reverse micelles and microemulsions of cetyltrimethylammonium bromide

1970 ◽  
Vol 24 (2) ◽  
pp. 173-184 ◽  
Author(s):  
Ferdousi Begum ◽  
Md Yousuf A Molla ◽  
M Muhibur Rahman ◽  
Md Abu Bin Hasan Susan
Keyword(s):  
Crystal Violet ◽  
Alkaline Hydrolysis ◽  
Reverse Micelles ◽  
Cetyltrimethylammonium Bromide ◽  
Specific Conductivity ◽  
Water System ◽  
Chemical Society ◽  
Order Rate Constant ◽  
Kinetics Of ◽  
Hydrolysis Of

Kinetics of the alkaline hydrolysis of crystal violet (CV) in micelles, reverse micelles and microemulsions of a cationic surfactant, cetyltrimethylammonium bromide (CTAB) was studied at 25 ± 0.1 oC using spectrophotometric method. The rate of alkaline hydrolysis of CV was catalyzed by micellar solutions of CTAB. The pseudo first order rate constant (k') has been found to decrease upon incorporation of 1-butanol to cationic CTAB micelles, which displaces the substrate from the micellar into the aqueous phase. In CTAB/cyclohexane/1-butanol/water system, as the content of 1-butanol increases, specific conductivity and density of the microemulsions and reverse micelles decrease. The change in physical properties also causes change in reaction environment. A change from a micelle-rich (o/w) to a reverse micelle-rich (w/o) condition is apparent for microemulsions and consequently the k' vs. % wt. of 1-butanol profiles show an initial decrease in the k' followed by a gradual increase and finally, to a sharp increase with increasing 1-butanol content. Microemulsions and reverse micelles thus offer the potential to control rate of a reaction by formation of micelles in water phase and reverse micelles in oil phase. DOI: http://dx.doi.org/10.3329/jbcs.v24i2.9706 Journal of Bangladesh Chemical Society, Vol. 24(2), 173-184, 2011

Kinetics of the hydrolysis of cellobiose and p-nitrophenyl-β-D-glucoside by cellobiase of Trichoderma viride

1977 ◽  
Vol 55 (1) ◽  
pp. 19-26 ◽  
Author(s):  
R. James Maguire
Keyword(s):  
Steady State ◽  
Trichoderma Viride ◽  
Solution Temperature ◽  
Temperature Range ◽  
A Value ◽  
Steady State Kinetics ◽  
Decreasing Ph ◽  
Ph Curve ◽  
Kinetics Of ◽  
Hydrolysis Of

Cellobiase has been isolated from the crude cellulase mixture of enzymes of Trichoderma viride using column chromatographic and ion-exchange methods. The steady-state kinetics of the hydrolysis of cellobiose have been investigated as a function of cellobiose and glucose concentrations, pH of the solution, temperature, and dielectric constant, using isopropanol–buffer mixtures. The results show that (i) there is a marked activation of the reaction by initial glucose concentrations of 4 × 10−3 M to 9 × 10−2 M and strong inhibition of the reaction at higher initial concentrations, (ii) the log rate – pH curve has a maximum at pH 5.2 and enzyme pK values of 3.5 and 6.8, (iii) the energy of activation at pH 5.1 is 10.2 kcal mol−1 over the temperature range 5–56 °C, and (iv) the rate decreases from 0 to 20% (v/v) isopropanol.The hydrolysis by cellobiase (EC 3.2.1.21) of p-nitrophenyl-β-D-glucoside was examined by pre-steady-state methods in which [Formula: see text], and by steady-state methods as a function of pH and temperature. The results show (i) a value for k2 of 21 s−1 at pH 7.0 (where k2 is the rate constant for the second step in the assumed two-intermediate mechanism [Formula: see text]) (ii) a log rate–pH curve, significantly different from that for hydrolysis of cellobiose, in which the rate increases with decreasing pH below pH 4.5, is constant in the region pH 4.5–6, and decreases above pH 6 (exhibiting an enzyme pK value of 7.3), and (iii) an activation energy of 12.5 kcal mol−1 at pH 5.7 over the temperature range 10–60 °C.

Untersuchung der alkalischen Hydrolyse des Benzoesäurebenzylesters in Wasser-Äthanol 1 : 1 / Study of the Basic Hydrolysis of Benzyl Benzoate in Water-Ethyl Alcohol 1:1

1974 ◽  
Vol 29 (11) ◽  
pp. 1697-1698 ◽  
Author(s):  
F. Mansilla ◽  
P. Martinez ◽  
J. Sancho
Keyword(s):  
Experimental Data ◽  
Reaction Mechanism ◽  
Order Rate Constant ◽  
Benzyl Benzoate ◽  
Conductometric Method ◽  
Alcohol Medium ◽  
Basic Hydrolysis ◽  
Kinetics Of ◽  
Hydrolysis Of ◽  
Water Alcohol

By using a conductometric method, the kinetics of the basic hydrolysis of benzyl benzoate in water-alcohol medium has been investigated. The second order rate constant follows the equation K = A exp {-E/RT} with A = 1.35·1010 l mol-1 min-1 and E = 14.5 kcal mol-1. A reaction mechanism is postulated, which is consistent with the experimental data.

scholarly journals Acid Hydrolysis of Bromazepam Catalyzed by Micelles, Reverse Micelles, and Microemulsions

2015 ◽  
Vol 2015 ◽  
pp. 1-10
Author(s):  
Ferdousi Begum ◽  
M. Yousuf A. Mollah ◽  
M. Muhibur Rahman ◽  
Md. Abu Bin Hasan Susan
Keyword(s):  
Physicochemical Properties ◽  
Acid Hydrolysis ◽  
Reverse Micelles ◽  
Cetyltrimethylammonium Bromide ◽  
Order Rate Constant ◽  
Excellent Correlation ◽  
Initial Decrease ◽  
Droplet Sizes ◽  
Kinetics Of ◽  
Hydrolysis Of

Kinetics of the acid hydrolysis of bromazepam (Bz) has been investigated in micelles, reverse micelles, and microemulcions of cetyltrimethylammonium bromide (CTAB) by spectrophotometric method. The rate of the acid hydrolysis of Bz was found to be enhanced both below and above the critical micelle concentration (CMC) of CTAB in aqueous solution. The pseudo-first-order rate constant (k′) shows an initial decrease for both low and high H+concentrations. With further increase in [CTAB], at low [H+], thek′attains an almost constant value, while, at high [H+], thek′passes through a maximum and then decreases. The kinetic data for catalysis by micelles of CTAB was interpreted with the pseudophase ion exchange (PIE) model. In CTAB/cyclohexane/1-butanol/water microemulsions, as the water to surfactant ratio (wo) increases, the physicochemical properties and droplet sizes of microemulsions significantly change and distinct changes in reaction environment can be marked. The rate of the hydrolysis reaction exhibits excellent correlation with the physicochemical properties and droplet sizes of the microemulsions and reverse micelles of CTAB. At [H+] = 0.001 M, in reverse micelles and microemulsions of CTAB, thek′of the acid hydrolysis of Bz decreases sharply followed by a slight increase with increasingwo.

Steady-State Kinetics of Enzyme Reactions in the Presence of Added Nucleophiles

1972 ◽  
Vol 50 (12) ◽  
pp. 1334-1359 ◽  
Author(s):  
Irwin Hinberg ◽  
Keith J. Laidler
Keyword(s):  
Specific Binding ◽  
Specific Binding Sites ◽  
Steady State Kinetics ◽  
Special Cases ◽  
Kinetics Of Formation ◽  
Catalyzed Reactions ◽  
Enzyme Catalyzed Reactions ◽  
Concentration Graphs ◽  
Kinetics Of ◽  
Hydrolysis Of

Many enzyme-catalyzed reactions, such as hydrolyses, give rise to two products P1 and P2 which are formed in different reaction steps. The second product P2 is frequently formed by hydrolysis of an intermediate such as an acyl-enzyme or a phosphoryl-enzyme. An alternative nucleophile N introduced into the system forms an additional product P3. The present paper is concerned with the kinetics of formation of P1, P2, and P3 in the presence of added nucleophiles. A number of alternative mechanisms are considered, and equations are derived for the rates of formation of the three products, and the Michaelis constant, as functions of nucleophile concentration. Graphs are presented showing the variations of these parameters with the concentration of N, for a variety of special cases. Special attention is given to the possibility of specific binding sites for the water and the nucleophile molecules.The data for a number of enzyme systems are discussed with reference to the treatment. For reactions catalyzed by alkaline phosphatase it is concluded that only one mechanism (mechanism VI) is consistent with the results.

scholarly journals Chorismate synthase. Pre-steady-state kinetics of phosphate release from 5-enolpyruvylshikimate 3-phosphate

1990 ◽  
Vol 265 (3) ◽  
pp. 899-902 ◽  
Author(s):  
T R Hawkes ◽  
T Lewis ◽  
J R Coggins ◽  
D M Mousdale ◽  
D J Lowe ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Steady State ◽  
Lag Phase ◽  
Chorismate Synthase ◽  
Phosphate Release ◽  
Rate Limiting Step ◽  
Limiting Step ◽  
Steady State Kinetics ◽  
Rate Limiting ◽  
Kinetics Of

The pre-steady-state kinetics of phosphate formation from 5-enolpyruvylshikimate 3-phosphate catalysed by Escherichia coli chorismate synthase (EC 4.6.1.4) were studied by a rapid-acid-quench technique at 25 degrees C at pH 7.5. No pre-steady-state ‘burst’ or ‘lag’ phase was observed, showing that phosphate is released concomitant with the rate-limiting step of the enzyme. The implications of this result for the mechanism of action of chorismate synthase are discussed.

Presteady-state kinetics of Bacillus 1,3–1,4-β-glucanase: binding and hydrolysis of a 4-methylumbelliferyl trisaccharide substrate

2001 ◽  
Vol 357 (1) ◽  
pp. 195-202
Author(s):  
Mireia ABEL ◽  
Antoni PLANAS ◽  
Ulla CHRISTENSEN
Keyword(s):  
Steady State ◽  
Rate Constant ◽  
Product Formation ◽  
Lag Phase ◽  
Wild Type ◽  
Induced Fit ◽  
Enzyme Substrate ◽  
Kinetics Of ◽  
Hydrolysis Of ◽  
Presteady State Kinetics

In the present study the first stopped-flow experiments performed on Bacillus 1,3–1,4-β-glucanases are reported. The presteady-state kinetics of the binding of 4-methylumbelliferyl 3-O-β-cellobiosyl-β-d-glucoside to the inactive mutant E134A, and the wild-type-catalysed hydrolysis of the same substrate, were studied by measuring changes in the fluorescence of bound substrate or 4-methylumbelliferone produced. The presteady-state traces all showed an initial lag phase followed by a fast monoexponential phase leading to equilibration (for binding to E134A) or to steady state product formation (for the wild-type reaction). The lag phase, with a rate constant of the order of 100s−1, was independent of the substrate concentration; apparently an induced-fit mechanism governs the formation of enzyme–substrate complexes. The concentration dependencies of the observed rate constant of the second presteady-state phase were analysed according to a number of reaction models. For the reaction of the wild-type enzyme, it is shown that the fast product formation observed before steady state is not due to a rate-determining deglycosylation step. A model that can explain the observed results involves, in addition to the induced fit, a conformational change of the productive ES complex into a form that binds a second substrate molecule in a non-productive mode.

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