Structure of Thermobifida fusca DyP-type peroxidase and activity towards Kraft lignin and lignin model compounds

DOI: 10.1016/j.abb.2016.02.019

Authors: Rahman Rahmanpour (University of Warwick) , Dean Rea (University of Warwick) , Shirin Jamshidi (University of Warwick) , Vilmos Fulop (University of Warwick) , Timothy D. H. Bugg (University of Warwick)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Archives Of Biochemistry And Biophysics , VOL 594 , PAGES 54–60

State: Published (Approved)
Published: March 2016
Diamond Proposal Number(s): 8359

Abstract: A Dyp-type peroxidase enzyme from thermophilic cellulose degrader Thermobifida fusca (TfuDyP) was investigated for catalytic ability towards lignin oxidation. TfuDyP was characterised kinetically against a range of phenolic substrates, and a compound I reaction intermediate was observed via pre-steady state kinetic analysis at λmax 404 nm. TfuDyP showed reactivity towards Kraft lignin, and was found to oxidise a β-aryl ether lignin model compound, forming an oxidised dimer. A crystal structure of TfuDyP was determined, to 1.8 Å resolution, which was found to contain a diatomic oxygen ligand bound to the heme centre, positioned close to active site residues Asp-203 and Arg-315. The structure contains two channels providing access to the heme cofactor for organic substrates and hydrogen peroxide. Site-directed mutant D203A showed no activity towards phenolic substrates, but reduced activity towards ABTS, while mutant R315Q showed no activity towards phenolic substrates, nor ABTS.

Journal Keywords: Dye decolorizing peroxidase; Dyp; Lignin oxidation; Protein crystallography

Diamond Keywords: Enzymes

Subject Areas: Chemistry, Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography

Added On: 25/02/2016 13:48

Discipline Tags:

Biotechnology Biochemistry Chemistry Structural biology Engineering & Technology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)