Kinetic study of the thermal inactivation of cholinesterase enzymes immobilized in solid matrices

Bromberg, A.; Marx, Sharon; Frishman, Gad

doi:10.1016/j.bbapap.2008.02.018

Cited by 24 publications

(25 citation statements)

References 49 publications

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“…Positive ΔH # values indicate the endothermic character of the inactivation process [38]. ΔG # and ΔH # decreased as the temperature increased (Table 4), suggesting the destabilisation of protease P45 at higher temperatures [44,50]. The ΔS # values have not presented a continuous behaviour, which could result from the difficult evaluation of system disorder in such a small temperature variation.…”

Section: Resultsmentioning

confidence: 99%

Kinetic Stability Modelling of Keratinolytic Protease P45: Influence of Temperature and Metal Ions

Daroit

Sant’Anna

Brandelli

2011

Appl Biochem Biotechnol

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The activity and kinetic stability of a keratinolytic subtilisin-like protease from Bacillus sp. P45 was investigated in 100 mM Tris-HCl buffer (pH 8.0; control) and in buffer with addition of Ca(2+) or Mg(2+) (1-10 mM), at different temperatures. Addition of 3 mM Ca(2+) or 4 mM Mg(2+) resulted in a 26% increment on enzyme activity towards azocasein when compared to the control (100%; without added Ca(2+) or Mg(2+)) at 55 °C. Optimal temperature for activity in the control (55 °C) was similar with Mg(2+); however, temperature optimum was increased to 60 °C with 3 mM Ca(2+), displaying an enhancement of 42% in comparison to the control at 55 °C. Stability of protease P45 in control buffer and with Mg(2+) addition was assayed at 40-50 °C, and at 55-62 °C with Ca(2+) addition. Data were fitted to six kinetic inactivation models, and a first-order equation was accepted as the best model to describe the inactivation of protease P45 with and without metal ions. The kinetic and thermodynamic parameters obtained showed the crucial role of calcium ions for enzyme stability. As biocatalyst stability is fundamental for commercial/industrial purposes, the stabilising effect of calcium could be exploited aiming the application of protease P45 in protein hydrolysis.

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Section: Resultsmentioning

confidence: 99%

Kinetic Stability Modelling of Keratinolytic Protease P45: Influence of Temperature and Metal Ions

Daroit

Sant’Anna

Brandelli

2011

Appl Biochem Biotechnol

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“…At temperatures above 52°C, the Gibbs energy reached completely different levels (Figure 3). This difference between the temperature ranges can also be seen from the values of the enthalpy (ΔH; kJ/mol) and entropy (ΔS; J/mol.K), which indicate that the susceptibility of the enzyme to thermal denaturation increases with an increase in temperature (Bromberg et al, 2008).…”

Section: Thermodynamic Denaturation Parametersmentioning

confidence: 94%

“…With respect to the immobilized enzyme, the value of E ad was higher in temperature phase 2 (52-70°C) for both the pH values assessed. Several authors have shown that chemical modifications of the surface can drastically change enzyme behavior with respect to its thermostability and other aspects of interest, due to conformational changes (Bromberg et al, 2008;Bhatti et al, 2007;Šikšnis et al, 1990). It must be emphasized that immobilization, especially on a strongly loaded support such as niobium, can alter the enzyme behavior or highlight phenomena imperceptible when the enzyme is dispersed in the medium (Sadana, 1998).…”

Section: B) Immobilized Enzymementioning

confidence: 99%

“…On the other hand, the operational stability describes the enzyme persistence during the process under the conditions of use (Papinutti et al, 2008;Ó'Fágáin, 2003). Immobilization also changes the denaturation profile in both negative and positive ways, one explanation for this being that the decrease in the solubility of the enzyme reduces, for instance, molecular and submolecular mobility and vibrations (Bromberg et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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Thermal stability of the immobilized fructosyltransferase from Rhodotorula sp

Aguiar‐Oliveira

Maugeri

2011

Braz. J. Chem. Eng.

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-The thermal stability of the extracellular fructosyltransferase (FTase) from Rhodotorula sp., recovered from cultivation medium by ethanol precipitation and immobilized onto niobium ore, was studied by Arrhenius plot, half-life profile, half-inactivation temperature (T 50 ) and thermodynamic parameters. The Arrhenius plot showed two different behaviors with different deactivation energies (E ad ) only after immobilization, the transition occurring in the temperature interval between 51 and 52°C. T 50 for the free enzyme was estimated to be around 62°C and, after immobilization, 66°C. After 15 minutes at 52°C, it was also possible to observe enzymatic activation for both the free and immobilized forms, but greater activation was achieved at pH 4.5 with the immobilized enzyme. Between 47-51°C the immobilized enzyme was more stable than the free enzyme, with pH 6.0 being the more stable condition for the immobilized enzyme. However, above 52°C the free form was more stable.

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“…Furthermore, the high values obtained for ΔS # variation probably reflect an increased disorder of the active site or the structure of each carboxylmethylcellulase, which is the main driving force of heat denaturation (D'amico et al, 2003). Generally, activation entropy has a dominant role in thermal inactivation of proteins in aqueous solutions (Bromberg et al, 2008).…”

Section: Thermodynamic Studies Of Carboxylmethylcellulasesmentioning

confidence: 99%

Kinetic and Thermodynamic Analysis of Two Carboxymethylcellulases from Macrotermes subhyalinus Little Soldier

Bedel¹,

Gnanwa²,

Kone³

et al. 2017

IJB

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Optimization of thermal processes relies on adequate degradation kinetic models to warrant food safety and quality. The knowledge on thermal inactivation of enzymes is necessary to allow their proper utilization in food industry and technology applications, enabling the reduction of heating times and optimization of heating temperatures. In this work, the kinetic of thermal inactivation was studied for the previously characterized carboxylmethylcellulases Ab-CX1 and Ab-CX2 from Macrotermes subhyalinus little soldier. Samples of carboxymethylcellulases were treated at different time-temperature combinations in the range of 5-60 min at 50-65°C and the kinetic and thermodynamic parameters for carboxymethylcellulases were calculated. Results showed that inactivation followed a first-order reaction with k-values between 0.0103 ± 0.0003 to 0.1217 ± 0.0005 and 0.0149 ± 0.0007 to 0.0416 ± 0.0003 min -1 for Ab-CX1 and Ab-CX2, respectively. At high temperatures, Ab-CX2 was less resistant, with a significant decrease in residual activity compared to Ab-CX1. The D-and k-values decreased and increased, respectively, with increasing temperature, indicating faster inactivation of carboxymethylcellulases. Activation energy (Ea) and Z-values were estimated to 76.74 ± 1.98 kJ.mol were also calculated. The high value obtained for the variation in enthalpy of activation indicates that a high amount of energy is required to initiate denaturation, probably due to the molecular conformation of carboxymethylcellulases. All results suggest that both carboxymethylcellulases are relatively resistant to long heat treatments up to 50°C.

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Kinetic study of the thermal inactivation of cholinesterase enzymes immobilized in solid matrices

Cited by 24 publications

References 49 publications

Kinetic Stability Modelling of Keratinolytic Protease P45: Influence of Temperature and Metal Ions

Kinetic Stability Modelling of Keratinolytic Protease P45: Influence of Temperature and Metal Ions

Thermal stability of the immobilized fructosyltransferase from Rhodotorula sp

Kinetic and Thermodynamic Analysis of Two Carboxymethylcellulases from Macrotermes subhyalinus Little Soldier

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