Kinetic and thermodynamic characterization of the functional properties of a hybrid versatile peroxidase using isothermal titration calorimetry: Insight into manganese peroxidase activation and lignin peroxidase inhibition

Biochimie
Haluk ErtanRicardo Cavicchioli

Abstract

Isothermal titration calorimetry (ITC) was developed for measuring lignin peroxidase (LiP) and manganese peroxidase (MnP) activities of versatile peroxidase (VP) from Bjerkandera adusta. Developing an ITC approach provided an alternative to colorimetric methods that enabled reaction kinetics to be accurately determined. Although VP from Bjerkandera adjusta is a hybrid enzyme, specific conditions of [Mn+2] and pH were defined that limited activity to either LiP or MnP activities, or enabled both to be active simultaneously. MnP activity was found to be more efficient than LiP activity, with activity increasing with increasing concentrations of Mn+2. These properties of MnP were explained by a second metal binding site involved in homotropic substrate (Mn+2) activation. The activation of MnP was also accompanied by a decrease in both activation energy and substrate (Mn) affinity, reflecting a flexible enzyme structure. In contrast to MnP activity, LiP activity was inhibited by high dye (substrate) concentrations arising from uncompetitive substrate inhibition caused by substrate binding to a site distinct from the catalytic site. Our study provides a new level of understanding about the mechanism of substrate regulation of cataly...Continue Reading

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Citations

Mar 26, 2015·Plant Science : an International Journal of Experimental Plant Biology·Gea GuerrieroKhawar Sohail Siddiqui
Oct 3, 2012·Protein Expression and Purification·Oliver SpadiutChristoph Herwig
Jan 28, 2017·Metallomics : Integrated Biometal Science·Cristina N Butterfield, Bradley M Tebo
Feb 28, 2018·Journal of Molecular Microbiology and Biotechnology·Nancy Coconi LinaresMiguel A Gómez-Lim
May 1, 2016·Applied and Environmental Microbiology·Doriv KnopYitzhak Hadar
Nov 17, 2020·Frontiers in Molecular Biosciences·Yun WangAnthony K Mittermaier

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