scholarly journals Mechanism of acylphosphatase inactivation by Woodward's reagent K

1997 ◽  
Vol 328 (3) ◽  
pp. 855-861 ◽  
Author(s):  
Paolo PAOLI ◽  
Tania FIASCHI ◽  
Paolo CIRRI ◽  
Guido CAMICI ◽  
Giampaolo MANAO ◽  
...  
Keyword(s):  
Active Site ◽  
Kinetic Constant ◽  
Enzyme Inactivation ◽  
High Catalytic Activity ◽  
Order Kinetic ◽  
Recombinant Enzymes ◽  
Strong Absorption Band ◽  
First Order ◽  
Woodward’S Reagent K ◽  
Pseudo First Order

The organ common-type (CT) isoenzyme of acylphosphatase is inactivated by Woodward's reagent K (WRK) (N-ethyl-5-phenylisoxazolium-3ʹ-sulphonate) at pH 6.0. The inactivation reaction follows apparent pseudo first-order kinetics. The dependence of the reciprocal of the pseudo first-order kinetic constant (kobs) on the reciprocal WRK concentration reveals saturation kinetics, suggesting that the WRK forms a reversible complex with the enzyme before causing inactivation. Competitive inhibitors, such as inorganic phosphate and ATP, protect the enzyme from WRK inactivation, suggesting that this reagent acts at or near to the enzyme active site. The reagent-enzyme adduct, which elicits a strong absorption band with λmax at 346 nm, was separated from unreacted enzyme by reverse phase HPLC and the modified protein was cleaved with endoproteinase Glu-C to produce fragments. The HPLC fractionation gave two reagent-labelled peptides (peak 1 and peak 2) that were analysed by ion-spray MS and sequenced. The former is VFFRKHTQAE (residues 20-29 of human CT acylphosphatase) and the latter IFGKVQGVFFRKHTQAE (residues 13-29). MS demonstrated that both peptides are WRK adducts. A fragment ion with m/z of 1171, which is present in the mass spectrum of peak 1, has been identified as a WRK adduct of the peptide fragment 20-26. The λmax at 346 nm of WRK adduct suggests that the modified residue is His-25. Five recombinant enzymes mutated in residues included in the 20-29 polypeptide stretch have been produced. Analysis of their reactivities with WRK demonstrates that His-25 is the molecular target of the reagent as its modification causes the inactivation of the enzyme. Since both His-25 → Gln and His-25 → Phe mutants maintain high catalytic activity, we suggest that the observed enzyme inactivation is caused by the reagent (covalently bound to His-25), which shields the active site.

The Influence of Boria and EDTA on the Hydrodesulfurization Activity on CoMoS Supported Catalysts

2011 ◽  
Vol 9 (1) ◽  
Author(s):  
Matin Parvari ◽  
Peyman Moradi
Keyword(s):  
Ethylenediaminetetraacetic Acid ◽  
Supported Catalysts ◽  
Batch Reactor ◽  
Kinetic Constant ◽  
Order Kinetic ◽  
First Order ◽  
Pseudo First Order ◽  
Xrd Studies ◽  
Al2o3 Catalyst ◽  
Hydrodesulfurization Activity

The hydrodesulfurization of dibenzothiophene (HDS of DBT) in a high pressure batch reactor at 320°C was carried out over CoMo/Al2O3-B2O3 catalysts with different B2O3 contents (4, 10, and 16 wt%). Ethylenediaminetetraacetic acid (EDTA) with different EDTA/Co mole ratios (0.6, 1.2 and 1.8) was used as a chelating ligand during the preparation of CoMo/Al2O3-B2O3. XRD studies, FTIR, TPD of NH3, and BET experiments were used to investigate the catalyst samples. The results showed that the catalyst using the support with 4 wt% B2O3 and an EDTA/Co mole ratio of 1.2 had a hydrodesulfurization activity (in pseudo first order kinetic constant basis) value of ~2.96 times higher than that of the simple CoMo/Al2O3 catalyst.

scholarly journals Ganglioside incorporation and release by the isolated perfused rat liver

1991 ◽  
Vol 274 (2) ◽  
pp. 581-585 ◽  
Author(s):  
S C Kivatinitz ◽  
A Miglio ◽  
R Ghidoni
Keyword(s):  
Rat Liver ◽  
The Other ◽  
Regulatory Role ◽  
Isolated Perfused Rat Liver ◽  
Order Kinetic ◽  
Perfused Rat Liver ◽  
Ganglioside Gm1 ◽  
First Order ◽  
Pseudo First Order

The fate of exogenous ganglioside GM1 labelled in the sphingosine moiety, [Sph-3H]GM1, administered as a pulse, in the isolated perfused rat liver was investigated. When a non-recirculating protocol was employed, the amount of radioactivity in the liver and perfusates was found to be dependent on the presence of BSA in the perfusion liquid and on the time elapsed after the administration of the ganglioside. When BSA was added to the perfusion liquid, less radioactivity was found in the liver and more in the perfusate at each time tested, for up to 1 h. The recovery of radioactivity in the perfusates followed a complex course which can be described by three pseudo-first-order kinetic constants. The constants, in order of decreasing velocity, are interpreted as: (a) the dilution of the labelled GM1 by the constant influx of perfusion liquid; (b) the washing off of GM1 loosely bound to the surface of liver cells; (c) the release of gangliosides from the liver. Process (b) was found to be faster in the presence of BSA, probably owing to the ability of BSA to bind gangliosides. The [Sph-3H]GM1 in the liver underwent metabolism, leading to the appearance of products of anabolic (GD1a, GD1b) and catabolic (GM2, GM3) origin; GD1a appeared before GM2 and GM3 but, at times longer than 10 min, GM2 and GM3 showed more radioactivity than GD1a. At a given time the distribution of the radioactivity in the perfusates was quite different from that of the liver. In fact, after 60 min GD1a was the only metabolite present in any amount, the other being GM3, the quantity of which was small. This indicates that the liver is able to release newly synthesized gangliosides quite specifically. When a recirculating protocol was used, there were more catabolites and less GD1a than with the non-recirculating protocol. A possible regulatory role of ganglioside re-internalization on their own metabolism in the liver is postulated.

scholarly journals Enhanced photocatalytic activity of wool-ball-like TiO2 microspheres on carbon fabric and FTO substrates

2018 ◽  
Vol 38 ◽  
pp. 02014
Author(s):  
Yu Zhang ◽  
Jian Gu ◽  
Mengqi Zhang
Keyword(s):  
Hydrothermal Method ◽  
Order Kinetic ◽  
Carbon Fabric ◽  
Facile Hydrothermal Method ◽  
Sem Images ◽  
First Order ◽  
Good Crystallinity ◽  
Degradation Rate Constant ◽  
Order Kinetic Model ◽  
Pseudo First Order

The wool-ball-like TiO2 microspheres on carbon fabric (TiO2-CF) and FTO substrates (TiO2-FTO) have been synthesized by a facile hydrothermal method in alkali environment, using commercial TiO2 (P25) as precursors. The XRD results indicate that the as-prepared TiO2 have good crystallinity. And the SEM images show that the wool-ball-like TiO2 microspheres with a diameter of 2-3 μm are composed of TiO2 nanowires, which have a diameter of ~50 nm. The photocatalytic behavior of the wool-ball-like TiO2 microspheres, TiO2-CF and TiO2-FTO under ultraviolet light was investigated by a pseudo first-order kinetic model, using methyl orange (MO) as pollutant. The wool-ball-like TiO2 microspheres obtained a degradation rate constant (Kap) of 6.91×10-3 min-1 . The Kap values of TiO2-FTO and TiO2-CF reach 13.97×10-3 min-1 and 11.80×10-3 min-1, which are 2.0 and 1.7 times higher than that of pristine wool-ball-like TiO2 microspheres due to the “sum effect” between TiO2 and substrates. This study offers a facile hydrothermal method to prepare wool-ball-like TiO2 microspheres on CF and FTO substrates, which will improve the recyclability of phtocatalysts and can be extended to other fields.

scholarly journals Photocatalytic degradation of indigo carmine by ZnO photocatalyst under visible light irradiation

2017 ◽  
Vol 14 (3) ◽  
pp. 582-587
Author(s):  
Baghdad Science Journal
Keyword(s):  
Zinc Oxide ◽  
Photocatalytic Degradation ◽  
Indigo Carmine ◽  
Catalyst Loading ◽  
Order Kinetic ◽  
Kinetic Reaction ◽  
First Order ◽  
Pseudo First Order ◽  
The Relationship ◽  
Influential Parameters

In this work, the photocatalytic degradation of indigo carmine (IC) using zinc oxide suspension was studied. The effect of influential parameters such as initial indigo carmine concentration and catalyst loading were studied with the effect of Vis irradiation in the presence of reused ZnO was also investigated. The increased in initial dye concentration decreased the photodegradation and the increased catalyst loading increased the degradation percentage and the reused-ZnO exhibits lower photocatalytic activity than the ZnO catalyst. It has been found that the photocatalytic degradation of indigo carmine obeyed the pseudo-first-order kinetic reaction in presence of zinc oxide. This was found from plotting the relationship between ln (C0/Ct) and irradiation the rate constant of the process.UV- spectrophotometer was used to study the indigo carmine photodegradation.

Degradation of 4-nitrophenol using the Fenton process

2000 ◽  
Vol 42 (3-4) ◽  
pp. 155-160 ◽  
Author(s):  
Y.-S. Ma ◽  
S.-T. Huang ◽  
J.-G. Lin
Keyword(s):  
Kinetic Equation ◽  
Nitrogen Concentration ◽  
Order Kinetic ◽  
Fenton Process ◽  
Nitrogenous Compounds ◽  
First Order ◽  
Degradation Rate Constant ◽  
Before And After ◽  
Pseudo First Order ◽  
Order Kinetic Equation

In this study, The Fenton process was applied as a pretreatment method to treat industrial wastewater containing 4-nitrophenol (4-NP). The effect of oxidant dosages on the decomposition of 4-NP and the reaction kinetics were investigated. More than 99% of 4-NP was readily decomposed when the reaction was carried out at oxidant concentrations of 5 mM H2O2 and 5 mg/L Fe2+ for 2 hours. The total nitrogenous compounds and the nitrogen gas evolved, accounted for 88% of the initial nitrogen concentration. However, the maximum DOC removal efficiency was 30.6%; and only 1/3 of 4-NP was mineralized to carbon dioxide by the Fenton process. 4-NP degradation profiles fitted well into a pseudo first-order kinetic equation; degradation rate constant (min-1) of 4-NP increased from 4.3×10-3 to 66.1×10-3 with increasing dosages of H2O2 and Fe2+. In addition, the t value was calculated for studying the significance of simulation by the t-test. It was found that the t value was greater than the value for 99% confidence. This result suggested that the 4-NP decomposition profile could be described by the pseudo first-order kinetic equation. Biodegradability of 4-NP before and after the reaction was 0.018 and 0.594, respectively.

Evidence for the formation of lignin-hexenuronic acid-xylan complexes during modified kraft pulping processes

Holzforschung ◽  
2006 ◽  
Vol 60 (2) ◽  
pp. 137-142 ◽  
Author(s):  
Zhi-Hua Jiang ◽  
Jean Bouchard ◽  
Richard Berry
Keyword(s):  
Acid Hydrolysis ◽  
Kraft Pulp ◽  
Kraft Pulping ◽  
Order Kinetic ◽  
Kraft Pulps ◽  
First Order ◽  
Pseudo First Order ◽  
Hexenuronic Acid ◽  
Kinetics Of ◽  
Hydrolysis Of

Abstract The finding that hexenuronic acid (HexA) groups can be selectively removed from kraft pulps by acid hydrolysis has provided an opportunity to reduce bleaching chemicals. However, there is evidence that the acid hydrolysis is not uniform. In this report, we evaluate the kinetics of acid hydrolysis of HexA in a xylan sample enriched with HexA, a conventional kraft pulp, and three modified kraft pulps: anthraquinone pulp (Kraft-AQ), polysulfide pulp (PS), and polysulfide-anthraquinone pulp (PS-AQ). We found that HexA present in the xylan and conventional kraft pulp behaved similarly toward the acid hydrolysis throughout. On the other hand, HexA present in the Kraft-AQ, PS-AQ and PS pulps was heterogeneous toward acid hydrolysis and the reaction can be separated into two pseudo-first-order kinetic phases, each of which has a different rate constant. The kinetic data provide evidence for the formation of lignin-HexA-xylan complexes during modified kraft pulping processes.

scholarly journals Acrolein, an irreversible active-site-directed inhibitor of deoxyribose 5-phosphate aldolase?

1976 ◽  
Vol 153 (2) ◽  
pp. 495-497 ◽  
Author(s):  
D C Wilton
Keyword(s):  
Schiff Base ◽  
Rate Constant ◽  
Active Site ◽  
Order Rate Constant ◽  
Lysine Residue ◽  
Prolonged Incubation ◽  
Enzyme Active Site ◽  
First Order ◽  
Substrate Analogue ◽  
Pseudo First Order

The enzyme deoxyribose 5-phosphate aldolase was irreversibly inactivated by the substrate analogue acrolein with a pseudo-first-order rate constant of 0.324 min-1 and a Ki (apparent) of 2.7 × 10(-4) m. No inactivation was observed after prolonged incubation with the epoxide analogues glycidol phosphate and glycidaldehyde. It is suggested that the acrolein is first activated by forming a Schiff base with the enzyme active-site lysine residue and it is the activated inhibitor that reacts with a suitable-active-site nucleophile.

scholarly journals Probing the active site residues in aromatic donor oxidation in horseradish peroxidase: involvement of an arginine and a tyrosine residue in aromatic donor binding

1996 ◽  
Vol 314 (3) ◽  
pp. 985-991 ◽  
Author(s):  
Subrata ADAK ◽  
Abhijit MAZUMDER ◽  
Ranajit K. BANERJEE
Keyword(s):  
Active Site ◽  
Native Enzyme ◽  
Tyrosine Residue ◽  
Tyrosine Residues ◽  
First Order ◽  
First Order Kinetics ◽  
Aromatic Donor ◽  
Pseudo First Order ◽  
Modified Enzyme ◽  
Compound Ii

The plausible role of arginine and tyrosine residues at the active site of horseradish peroxidase (HRP) in aromatic donor (guaiacol) oxidation was probed by chemical modification followed by characterization of the modified enzyme. The arginine-specific reagents phenylglyoxal (PGO), 2,3-butanedione and 1,2-cyclohexanedione all inactivated the enzyme, following pseudo-first-order kinetics with second-order rate constants of 24 M-1·min-1, 0.8 M-1·min-1 and 0.54 M-1·min-1 respectively. Modification with tetranitromethane, a tyrosine-specific reagent, also resulted in 50% loss of activity following pseudo-first-order kinetics with a second-order rate constant of 2.0 M-1·min-1. The substrate, H2O2, and electron donors such as I- and SCN- offered no protection against inactivation by both types of modifier, whereas the enzyme was completely protected by guaiacol or o-dianisidine, an aromatic electron donor (second substrate) oxidized by the enzyme. These studies indicate the involvement of arginine and tyrosine residues at the aromatic donor site of HRP. The guaiacol-protected phenylglyoxal-modified enzyme showed almost the same binding parameter (Kd) as the native enzyme, and a similar free energy change (∆G´) for the binding of the donor. Stoicheiometric studies with [7-14C]phenylglyoxal showed incorporation of 2 mol of phenylglyoxal per mol of enzyme, indicating modification of one arginine residue for complete inactivation. The difference absorption spectrum of the tetranitromethane-modified against the native enzyme showed a peak at 428 nm, characteristic of the nitrotyrosyl residue, that was abolished by treatment with sodium dithionite, indicating specific modification of a tyrosine residue. Inactivation stoicheiometry showed that modification of one tyrosine residue per enzyme caused 50% inactivation. Binding studies by optical difference spectroscopy indicated that the arginine-modified enzyme could not bind guaiacol at all, whereas the tyrosine-modified enzyme bound it with reduced affinity (Kd 35 mM compared with 10 mM for the native enzyme). Both the modified enzymes, however, retained the property of the formation of compound II (one-electron oxidation state higher than native ferriperoxidase) with H2O2, but reduction of compound II to native enzyme by guaiacol did not occur in the PGO-modified enzyme, owing to lack of binding. No non-specific change in protein structure due to modification was evident from circular dichroism studies. We therefore suggest that the active site of HRP for aromatic donor oxidation is composed of an arginine and an adjacent tyrosine residue, of which the former plays an obligatory role in aromatic donor binding whereas the latter residue plays a facilitatory role, presumably by hydrophobic interaction or hydrogen bonding.

scholarly journals Adsorption of Cu (II) and Ni (II) Ions from Solution onto Calcium Alginate Beads

2020 ◽  
Vol 24 (2) ◽  
pp. 329-333
Author(s):  
D.O. Jalija ◽  
A . Uzairu
Keyword(s):  
Contact Time ◽  
Calcium Alginate ◽  
Alginate Beads ◽  
Second Order ◽  
Ion Concentration ◽  
Order Kinetic ◽  
Calcium Alginate Beads ◽  
First Order ◽  
Pseudo Second Order ◽  
Pseudo First Order

The objective of this study was to investigate the biosorption of Cu (II) and Ni (II) ions from aqueous solution by calcium alginate beads. The effects of solution pH, contact time and initial metal ion concentration were evaluated. The results showed that maximum Cu (II) removal (93.10%) occurred at pH of 9.0, contact time of 120 minutes and initial ion concentration of 10 mg/L while that of Ni (II) was 94.6%, which was achieved at pH of 8.0, contact time of 120 minutes and initial ion concentration of 10 mg/L. The equilibrium data fitted well to the Langmuir Isotherm indicating that the process is a monolayer adsorption. The coefficients of determination, R2, values for the Langmuir Isotherm were 0.9799 and 0.9822 respectively for Cu (II) and Ni (II) ions. The values of the maximum biosorption capacity, Qo, were 10.79 and 6.25 mgg-1 respectively. The kinetic data also revealed that the sorption process could best be described by the pseudo – second order kinetic model. The R2 values for the pseudo – second order kinetic plots for Cu (II) and Ni (II) were 0.9988 and 0.9969 respectively. These values were higher than those for the pseudo – first order plots. The values of the biosorption capacity qe obtained from the pseudo – second order plots were very close to the experimental values of qe indicating that the biosorption process follows the second order kinetics. This study has therefore shown that calcium alginate beads can be used for the removal of Cu (II) and Ni (II) ions from wastewaters. Keywords: Keywords: Adsorption, Calcium alginate, Isotherm, Langmuir, Pseudo- first order, Pseudo-second order

Sustainable wastewater management technology for tourism in Thailand: case and experimental studies

2019 ◽  
Vol 79 (10) ◽  
pp. 1977-1984
Author(s):  
W. Liamlaem ◽  
L. Benjawan ◽  
C. Polprasert
Keyword(s):  
Kinetic Constant ◽  
Experimental Studies ◽  
Oxygen Demand ◽  
Domestic Wastewater ◽  
Wastewater Management ◽  
Order Kinetic ◽  
First Order ◽  
Treated Effluent ◽  
Management Technology ◽  
Pre Treatment

Abstract Thailand has adopted the concept of eco-tourism as a protocol to protect environmental resources. One of the key factors in enabling the achievement of this goal is the improvement of the quality of effluent from those homestays and resorts which still lack efficient on-site wastewater treatment. This research utilized case studies of subsurface flow constructed wetlands (SFCWs), planted mainly with the Indian shot (Canna indica L.), which were designed to treat wastewaters at three resorts located in Amphawa District, Samut Songkram Province in central Thailand. The results showed that the treated effluent was of sufficient quality to meet the building effluent standards Type C, which require the concentrations of biological oxygen demand (BOD), total Kjeldahl nitrogen (TKN) and suspended solids (SS) to be less than 40, 40 and 50 mg/L, respectively. In addition, the first-order kinetic constants for the design and operation of SFCWs were determined. For treating wastewater containing organic substances, with no prior pre-treatment, the first-order kinetic constant of 0.24 1/d can be applied to predict effluent quality. For treating other types of domestic wastewater, a first-order kinetic constant in the range 0.40–0.45 1/d can be used when sizing and operating SFCWs. This research highlights the great potential of SFCWs as a sustainable wastewater management technology.

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