Kinetics and thermodynamics of thermal inactivation for recombinant Escherichia coli cellulases, cel12B, cel8C, and polygalacturonase, peh28; biocatalysts for biofuel precursor production

Authors

Eman Ibrahim
Ahmed Mahmoud, The University of Texas Rio Grande Valley
Kim D. Jones
Keith E. Taylor
Ebtesam N. Hosseney
Patrick L. Mills
Jean M. Escudero

Document Type

Article

Publication Date

1-2021

Abstract

Lignocellulosic biomass conversion using cellulases/polygalacturonases is a process that can be progressively influenced by several determinants involved in cellulose microfibril degradation. The current paper focuses on the kinetics and thermodynamics of thermal inactivation of recombinant E. coli cellulases, cel12B, cel8C, and a polygalacturonase, peh 28, derived from Pectobacterium carotovorum sub sp. carotovorum. Several consensus motifs conferring the enzymes’ thermal stability in both cel12B and peh28 model structures have been detailed earlier, which were confirmed for the three enzymes through the current study of their thermal inactivation profiles over the 20-80 °C range using the respective activities on carboxymethylcellulose and polygalacturonic acid. Kinetic constants and half-lives of thermal inactivation, inactivation energy, plus inactivation entropies, enthalpies and Gibbs free energies, revealed high stability, less conformational change and protein unfolding for cel12B and peh28 due to thermal denaturation compared to cel8C. The apparent thermal stability of peh28 and cel12B, along with their hydrolytic efficiency on a lignocellulosic biomass conversion as reported previously, makes these enzymes candidates for various industrial applications. Analysis of the Gibbs free energy values suggests that the thermal stabilities of cel12B and peh28 are entropy-controlled over the tested temperature range.

Comments

© The Author(s) 2020. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved. Original published version available at https://doi.org/10.1093/jb/mvaa097

Recommended Citation

Ibrahim, Eman, Ahmed Mahmoud, Kim D. Jones, Keith E. Taylor, Ebtesam N. Hosseney, Patrick L. Mills, and Jean M. Escudero. 2021. “Kinetics and Thermodynamics of Thermal Inactivation for Recombinant Escherichia Coli Cellulases, Cel12B, Cel8C, and Polygalacturonase, Peh28; Biocatalysts for Biofuel Precursor Production.” Journal of Biochemistry 169 (1): 109–17. https://doi.org/10.1093/jb/mvaa097.

Publication Title

The Journal of Biochemistry

DOI

10.1093/jb/mvaa097