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Thermodynamics and Kinetics of Heat Inactivation of a Novel Keratinase from Chryseobacterium sp. Strain kr6

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Abstract

A novel keratinase from Chryseobacterium sp. strain kr6 was purified to homogeneity by (NH4)2SO4 precipitation, gel permeation on Sephadex G-100, and Q-Sepharose Fast Flow anion-exchange chromatography. The molecular weight of the purified enzyme was around 20 kDa. Kinetic and thermodynamic parameters for thermal inactivation were determined. The influence of Ca2+ and Mg2+ ions and purification degree on the enzyme stability was evaluated in the range of 50 to 60 °C. The results showed that first-order kinetics explained well the thermal denaturation of the keratinase in this temperature interval. The presence of Ca2+ increases significantly the enzyme stability. Compared with the controls, the half-life of the purified enzyme after two purification steps in the presence of Ca2+ increased 7.3, 20.2, and 9.8 fold at 50, 55, and 60 °C, respectively. Thermodynamics parameters for thermal inactivation were also determined.

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Abbreviations

C 0 :

Activity at time t = 0

C t :

Activity at time t

k d :

First-order rate constant (min−1)

E a,d :

Activation energy for denaturation (kJ mol−1)

ΔG #d :

Free energy (kJ mol−1)

ΔH #d :

Activation enthalpy (kJ mol−1)

ΔS #d :

Activation entropy (J mol−1 K−1)

H :

Planck constant (J s)

K B :

Boltzmann constant (J K−1)

R :

Universal gas constant (J mol−1 K−1)

r 2 :

Determination coefficient

t :

Time (min)

t 1/2 :

Half-life time (min)

T :

Temperature (K)

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Acknowledgements

This work was supported by CAPES and CNPq (Brazil).

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Authors and Affiliations

  1. Laboratório de Bioquímica e Microbiologia Aplicada, Departamento de Ciência de Alimentos, Universidade Federal do Rio Grande do Sul (UFRGS), Av. Bento Gonçalves 9500, 91501-970, Porto Alegre, Brazil

    Silvana Terra Silveira, Franciani Casarin, Sabrine Gemelli & Adriano Brandelli

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  1. Silvana Terra Silveira
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  2. Franciani Casarin
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  3. Sabrine Gemelli
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  4. Adriano Brandelli
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Correspondence to Adriano Brandelli.

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Silveira, S.T., Casarin, F., Gemelli, S. et al. Thermodynamics and Kinetics of Heat Inactivation of a Novel Keratinase from Chryseobacterium sp. Strain kr6. Appl Biochem Biotechnol 162, 548–560 (2010). https://doi.org/10.1007/s12010-009-8835-1

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