scholarly journals Purification and characterization of quinoprotein glucose dehydrogenase from Acinetobacter calcoaceticus L.M.D. 79.41

1986 ◽  
Vol 239 (1) ◽  
pp. 163-167 ◽  
Author(s):  
P Dokter ◽  
J Frank ◽  
J A Duine
Keyword(s):  
Electron Acceptor ◽  
Glucose Dehydrogenase ◽  
Substrate Inhibition ◽  
Acinetobacter Calcoaceticus ◽  
Reaction Rates ◽  
Reduced Form ◽  
Initial Reaction ◽  
Type I ◽  
Ph Optimum ◽  
Enzyme Form

Quinoprotein glucose dehydrogenase (EC 1.1.99.17) from Acinetobacter calcoaceticus L.M.D. 79.41 was purified to homogeneity. It is a basic protein with an isoelectric point of 9.5 and an Mr of 94,000. Denaturation yields two molecules of PQQ/molecule and a protein with an Mr of 48000, indicating that the enzyme consists of two subunits, which are probably identical because even numbers of aromatic amino acids were found. The oxidized enzyme form has an absorption maximum at 350 nm, and the reduced form, obtained after the addition of glucose, at 338 nm. Since double-reciprocal plots of initial reaction rates with various concentrations of glucose or electron acceptor show parallel lines, and substrate inhibition is observed for glucose as well as for electron acceptor at high concentrations, a ping-pong kinetic behaviour with the two reactants exists. From the plots, Km values for glucose and Wurster's Blue of 22 mM and 0.78 mM respectively, and a Vmax. of 7.730 mumol of glucose oxidized/min per mg of protein were derived. The enzyme shows a broad substrate specificity for aldose sugars. Cationic electron acceptors are active in the assay, anionic acceptors are not. A pH optimum of 9.0 was found with Wurster's Blue and 6.0 with 2,6-dichlorophenol-indophenol. Two types of quinoprotein glucose dehydrogenases seem to exist: type I enzymes are acidic proteins from which PQQ can be removed by dialysis against EDTA-containing buffers (examples are found in Escherichia coli, Klebsiella aerogenes and Pseudomonas sp.); type II enzymes are basic proteins from which PQQ is not removed by dialysis against EDTA-containing buffers (examples are found in A. calcoaceticus and Gluconobacter oxydans).

scholarly journals Cytochrome b-562 from Acinetobacter calcoaceticus L.M.D. 79.41. Its characteristics and role as electron acceptor for quinoprotein glucose dehydrogenase

1988 ◽  
Vol 254 (1) ◽  
pp. 131-138 ◽  
Author(s):  
P Dokter ◽  
J E van Wielink ◽  
M A G van Kleef ◽  
J A Duine
Keyword(s):  
Cytochrome B ◽  
Electron Acceptor ◽  
Glucose Dehydrogenase ◽  
Acinetobacter Calcoaceticus ◽  
Primary Electron ◽  
Reduced Form ◽  
Alpha Band ◽  
Absorption Bands ◽  
Midpoint Potential ◽  
Triton X

A soluble cytochrome b was purified from Acinetobacter calcoaceticus L.M.D. 79.41. On the basis of the alpha-band maximum of a reduced preparation, measured at 25 degrees C, it is designated as cytochrome b-562. This cytochrome is a basic monomeric protein (pI 10.2; Mr 18,000), containing one protohaem group per molecule. The reduced form, at 25 degrees C, showed absorption bands at 428, 532 and 562 nm. At 77 K the alpha-band shifted to 560 nm (with a shoulder at 558 nm). The reduced cytochrome did not react with CO. Cytochrome b-562 is most probably (loosely) attached to the outside of the cytoplasmic membrane, since substantial amounts of it, equimolar to quinoprotein glucose dehydrogenase (GDH), were present in the culture medium when cells were grown in the presence of low concentrations of Triton X-100. The midpoint potential at pH 7.0 was found to be +170 mV, a value that was lowered to +145 mV by the presence of GDH. Since the GDH was shown to have a midpoint potential of +50 mV, cytochrome b-562 could function as the natural primary electron acceptor. Arguments to substantiate this view and to propose a role of ubiquinone-9 as electron acceptor for cytochrome b-562 are presented.

scholarly journals Urca nuclide production in Type-I X-ray bursts and implications for nuclear physics studies

2020 ◽  
Vol 500 (3) ◽  
pp. 2958-2968
Author(s):  
Grant Merz ◽  
Zach Meisel
Keyword(s):  
Light Curve ◽  
Nuclear Reaction ◽  
Nuclear Physics ◽  
Thermal Structure ◽  
Reaction Rates ◽  
High Mass ◽  
Type I ◽  
Model Calculations ◽  
X Ray ◽  
Lower Temperature

ABSTRACT The thermal structure of accreting neutron stars is affected by the presence of urca nuclei in the neutron star crust. Nuclear isobars harbouring urca nuclides can be produced in the ashes of Type I X-ray bursts, but the details of their production have not yet been explored. Using the code MESA, we investigate urca nuclide production in a one-dimensional model of Type I X-ray bursts using astrophysical conditions thought to resemble the source GS 1826-24. We find that high-mass (A ≥ 55) urca nuclei are primarily produced late in the X-ray burst, during hydrogen-burning freeze-out that corresponds to the tail of the burst light curve. The ∼0.4–0.6 GK temperature relevant for the nucleosynthesis of these urca nuclides is much lower than the ∼1 GK temperature most relevant for X-ray burst light curve impacts by nuclear reaction rates involving high-mass nuclides. The latter temperature is often assumed for nuclear physics studies. Therefore, our findings alter the excitation energy range of interest in compound nuclei for nuclear physics studies of urca nuclide production. We demonstrate that for some cases this will need to be considered in planning for nuclear physics experiments. Additionally, we show that the lower temperature range for urca nuclide production explains why variations of some nuclear reaction rates in model calculations impacts the burst light curve but not local features of the burst ashes.

Ensemble Learning Approach with LASSO for Predicting Catalytic Reaction Rates

Synlett ◽  
2020 ◽  
Author(s):  
Akira Yada ◽  
Kazuhiko Sato ◽  
Tarojiro Matsumura ◽  
Yasunobu Ando ◽  
Kenji Nagata ◽  
...  
Keyword(s):  
Ensemble Learning ◽  
Reaction Rates ◽  
Initial Reaction Rate ◽  
Training Dataset ◽  
Initial Reaction ◽  
Learning Approach ◽  
Learning Framework ◽  
Machine Learning Approach ◽  
Reasonable Prediction ◽  
Epoxidation Of Alkenes

AbstractThe prediction of the initial reaction rate in the tungsten-catalyzed epoxidation of alkenes by using a machine learning approach is demonstrated. The ensemble learning framework used in this study consists of random sampling with replacement from the training dataset, the construction of several predictive models (weak learners), and the combination of their outputs. This approach enables us to obtain a reasonable prediction model that avoids the problem of overfitting, even when analyzing a small dataset.

Poly(β-L-malate) hydrolase from Plasmodia of Physarum polycephalum

1995 ◽  
Vol 41 (13) ◽  
pp. 192-199 ◽  
Author(s):  
Christian Korherr ◽  
Michael Roth ◽  
Eggehard Holler
Keyword(s):  
Hydrophobic Interaction ◽  
Malic Acid ◽  
Physarum Polycephalum ◽  
Culture Medium ◽  
Specific Activity ◽  
Hydrophobic Interaction Chromatography ◽  
Reduced Form ◽  
Serine Esterase ◽  
Hydroxybutyric Acid ◽  
Ph Optimum

A 68-kDa extracellular glycoprotein from Physarum polycephalum that hydrolyses specifically poly(β-L-malic acid) by removing monomers of L-malic acid in an exolytic manner has been purified and characterized. The enzyme was purified 1740-fold from the culture medium by ammonium sulfate precipitation, hydrophobic interaction chromatography on butyl-Toyopearl, and gel permeation chromatography on Superdex 200 to a specific activity of 9.0 μmol∙min−1∙mg−1. The hydrolase was also purified from the cytosol, which contained 1 mg in 43 g cells in contrast to 1 mg extracellular enzyme in 28 L of culture medium. The pH optimum was pH 3.5 as a result of the effect of an acidic side chain on Vmax and the preferred binding of poly(β-L-malate) in the ionized form. Intracellular hydrolase was only marginally active on [14C]poly(β-L-malate) that had been injected into plasmodia. Poly(L-aspartate), poly(L-glutamate), poly(vinyl sulfate), and poly(acrylate) were neither bound nor degraded by the hydrolase. Poly(β-hydroxybutyric acid), which was considered the reduced form of poly(β-L-malate), was not a substrate. The enzyme is neither a metallo- nor a serine-esterase, and is distinct from poly(3-hydroxybutyric acid) depolymerases. It is related to a glucosidase with respect to hydrophobic interaction chromatography, the pH-activity dependence, and its inhibition with mercuribenzoate, N-bromosuccinimide, and D-gluconolactone, but not the use of the substrates.Key words: poly(β-L-malate), polymalatase, Physarum polycephalum, biodegradative polymer.

Effect of Substrate Inhibition and Cooperativity on the Electrochemical Responses of Glucose Dehydrogenase. Kinetic Characterization of Wild and Mutant Types

2011 ◽  
Vol 133 (32) ◽  
pp. 12801-12809 ◽  
Author(s):  
Fabien Durand ◽  
Benoît Limoges ◽  
Nicolas Mano ◽  
François Mavré ◽  
Rebeca Miranda-Castro ◽  
...  
Keyword(s):  
Glucose Dehydrogenase ◽  
Substrate Inhibition ◽  
Kinetic Characterization

scholarly journals Uptake of Zn, Pb, Cu and Fe Ions from Spent and Unspent Engine Oil Using Termite Soil

2017 ◽  
Vol 9 (3) ◽  
pp. 85
Author(s):  
Iwekumo Agbozu ◽  
Bassey Uwem ◽  
Boisa Ndokiari
Keyword(s):  
Contact Time ◽  
Diffusion Constant ◽  
Reaction Rates ◽  
Second Order ◽  
Engine Oil ◽  
Initial Reaction ◽  
Kinetics Equation ◽  
Spent Engine Oil ◽  
Pseudo Second Order ◽  
Fe Ions

Removal of Zn, Pb, Cu and Fe ions from unspent and spent engine oil was studied using Termite soil. Process parameters such as contact time and adsorbent dosage were varied. Values from contact time were used for predicting kinetics equation of their uptake. At optimum time of 40 minutes, percentage adsorption was of the order Fe>Zn>Cu>Pb for both spent and unspent engine oil. Kinetics equation such as Elovich, Intra-particle, Pseudo-first order and Pseudo-second order were tested. Results obtained shows that their sequestering pattern fit into the pseudo-second order kinetics. Initial reaction rates, h (mg/g.min) and α (mg. g-1min-1) for all metal ions obtained from Pseudo-second order and Elovich kinetic models followed the trends Zn>Fe>Cu>Pb and Zn>Fe>Pb>Cu respectively in spent engine oil while for unspent engine oil, the trend was Fe>Zn>Cu>Pb for h (mg/g.min) and Zn>Fe>Pb>Cu for α (mg. g-1min-1). Electrostatic attraction existing on the surface of the adsorbent assisted in the high initial reaction of Zn and Fe ions, implying good affinity of the ions for the adsorbent. Desorption constant ᵦ (g/mg) was of the trend Cu>Pb>Fe>Zn and Cu>Pb>Zn>Fe for spent and unspent engine oils respectively. Intra-particle diffusion constant kid (mgg-1min-1/2) followed a similar pattern, revealing strong binding between Zn and termite soil than any of the metal ion. This pilot research has been able to suggest a kinetic process for uptake of the studied ions from spent and unspent engine oil.

scholarly journals New Reaction Rates Of 64Ge(p,γ)65As(p,γ)66Se And The Impact On Type I X-ray Bursts

2017 ◽  
Author(s):  
Yi Hua Lam ◽  
Jianjun He ◽  
Hendrik Schatz ◽  
B. Alex Brown ◽  
Anuj Parikh
Keyword(s):  
Reaction Rates ◽  
Type I ◽  
X Ray ◽  
The Impact

scholarly journals Studies on Epoxidation of Tung oil with Hydrogen Peroxide Catalyzed by Sulfuric Acid

2020 ◽  
Vol 15 (3) ◽  
pp. 674-686
Author(s):  
Eni Budiyati ◽  
Rochmadi Rochmadi ◽  
Arief Budiman ◽  
Budhijanto Budhijanto
Keyword(s):  
Hydrogen Peroxide ◽  
Sulfuric Acid ◽  
Unsaturated Fatty Acids ◽  
Catalyst Concentration ◽  
Reaction Rates ◽  
Catalytic Reactions ◽  
Side Reactions ◽  
Initial Reaction ◽  
Tung Oil ◽  
Epoxidation Reaction

Tung oil with an iodine value (IV) of 99.63 g I2/100 g was epoxidized in-situ with glacial acetic acid and hydrogen peroxide (H2O2), in the presence sulfuric acid as catalyst. The objective of this research was to evaluate the effect of mole ratio of H2O2 to unsaturated fatty acids (UFA), reaction time and catalyst concentration in Tung oil epoxidation. The reaction kinetics were also studied. Epoxidation was carried out for 4 h. The reaction rates and side reactions were evaluated based on the IV and the conversion of the epoxidized Tung oil to oxirane. Catalytic reactions resulted in higher reaction rate than did non-catalytic reactions. Increasing the catalyst concentration resulted in a large decrease in the IV and an increase in the conversion to oxirane at the initial reaction stage. However, higher catalyst concentration in the epoxidation reaction caused to a decrease in reaction selectivity. The mole ratio of H2O2 to UFA had an influence identical to the catalyst concentration. The recommended optimum mole ratio and catalyst concentration in this study were 1.6 and 1.5%, respectively. The highest conversion was 48.94% for a mole ratio of 1.6. The proposed kinetic model provided good results and was suitable for all variations in reaction temperature. The activation energy (Ea) values were around 5.7663 to 76.2442 kcal/mol. Copyright © 2020 BCREC Group. All rights reserved 

scholarly journals Purification and characterization of two human erythrocyte arylamidases preferentially hydrolysing N-terminal arginine or lysine residues

1978 ◽  
Vol 175 (3) ◽  
pp. 1051-1067 ◽  
Author(s):  
K K Mäkinen ◽  
P L Mäkinen
Keyword(s):  
Substrate Inhibition ◽  
Gel Filtration ◽  
Deae Cellulose ◽  
Gel Permeation Chromatography ◽  
Ph Optimum ◽  
Preparative Scale ◽  
Molecular Weights ◽  
Gel Permeation ◽  
Lysine Residues ◽  
Enzyme I

Two arylamidases (I and II) were purified from human erythrocytes by a procedure that comprised removal of haemoglobin from disrupted cells with CM-Sephadex D-50, followed by treatment of the haemoglobin-free preparation subsequently with DEAE-cellulose, gel-permeation chromatography on Sephadex G-200, gradient solubilization on Celite, isoelectric focusing in a pH gradient from 4 to 6, gel-permeation chromatography on Sephadex G-100 (superfine), and finally affinity chromatography on Sepharose 4B covalently coupled to L-arginine. In preparative-scale purifications, enzymes I and II were separated at the second gel-permeation chromatography. Enzyme II was obtained as a homogeneous protein, as shown by several criteria. Enzyme I hydrolysed, with decreasing rates, the L-amino acid 2-naphtylamides of lysine, arginine, alanine, methionine, phenylalanine and leucine, and the reactions were slightly inhibited by 0.2 M-NaCl. Enzyme II hydrolysed most rapidly the corresponding derivatives of arginine, leucine, valine, methionine, proline and alanine, in that order, and the hydrolyses were strongly dependent on Cl-. The hydrolysis of these substrates proceeded rapidly at physiological Cl- concentration (0.15 M). The molecular weights (by gel filtration) of enzymes I and II were 85 000 and 52 500 respectively. The pH optimum was approx. 7.2 for both enzymes. The isoelectric point of enzyme II was approx. 4.8. Enzyme I was activated by Co2+, which did not affect enzyme II to any noticeable extent. The kinetics of reactions catalysed by enzyme I were characterized by strong substrate inhibition, but enzyme II was not inhibited by high substrate concentrations. The Cl- activated enzyme II also showed endopeptidase activity in hydrolysing bradykinin.

Enzymes involved in the metabolism of thiosulfate by Thiobacillus thioparus. II. Properties of adenosine-5′-phosphosulfate reductase

1970 ◽  
Vol 48 (3) ◽  
pp. 344-354 ◽  
Author(s):  
Ronald M. Lyric ◽  
Isamu Suzuki
Keyword(s):  
Molecular Weight ◽  
Cytochrome C ◽  
Electron Acceptor ◽  
Desulfovibrio Desulfuricans ◽  
Thiobacillus Denitrificans ◽  
Ph Optimum ◽  
Thiobacillus Thioparus ◽  
Aps Reductase

Adenosine-5′-phosphosulfate (APS) reductase was purified from Thiobacillus thioparus extracts 25- to 46-fold and the properties were studied. The molecular weight was 170 000 and the enzyme had 1 mole of FAD, 8–10 moles of iron, and 4–5 moles of labile sulfide. Cytochrome c as well as ferricyanide served as the electron acceptor. The pH optimum shifted from 7.4 to 9.5 when cytochrome c was used instead of ferricyanide. The Km values for sulfite and AMP were reduced from 2.5 mM and 100 μM to 17 μM and 2.5 μM, respectively, with cytochrome c as electron acceptor. Properties of the T. thioparus enzyme were compared to those of APS reductase isolated from Thiobacillus denitrificans and Desulfovibrio desulfuricans.

Sign in / Sign up

Export Citation Format

Share Document