Integrated experimental and theoretical investigation of copper active site properties of a lytic polysaccharide monooxygenase from Serratia marcescens

DOI: 10.1021/acs.inorgchem.4c00602

Authors: Alessia Munzone (Aix Marseille Univ, CNRS) , Manon Pujol (Aix Marseille Univ, CNRS) , Ashish Tamhankar (Max Planck Institute for Chemical Energy Conversion) , Chris Joseph (Max Planck Institute for Chemical Energy Conversion) , Ievgen Mazurenko (Aix Marseille Univ, CNRS) , Marius Réglier (Aix Marseille Univ, CNRS) , Sergio A. V. Jannuzzi (Max Planck Institute for Chemical Energy Conversion) , Antoine Royant (Université Grenoble Alpes, CNRS, CEA; European Synchrotron Radiation Facility) , Giuseppe Sicoli (LASIRE UMR CNRS 8516, Université de Lille) , Serena Debeer (Max Planck Institute for Chemical Energy Conversion) , Maylis Orio (Aix Marseille Univ, CNRS) , A. Jalila Simaan (Aix Marseille Univ, CNRS) , Christophe Decroos (Aix Marseille Univ, CNRS; Université de Strasbourg, CNRS, INSERM)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Inorganic Chemistry

State: Published (Approved)
Published: May 2024
Diamond Proposal Number(s): 31504

Abstract: In this paper, we employed a multidisciplinary approach, combining experimental techniques and density functional theory (DFT) calculations to elucidate key features of the copper coordination environment of the bacterial lytic polysaccharide monooxygenase (LPMO) from Serratia marcescens (SmAA10). The structure of the holo-enzyme was successfully obtained by X-ray crystallography. We then determined the copper(II) binding affinity using competing ligands and observed that the affinity of the histidine brace ligands for copper is significantly higher than previously described. UV–vis, advanced electron paramagnetic resonance (EPR), and X-ray absorption spectroscopy (XAS) techniques, including high-energy resolution fluorescence detected (HERFD) XAS, were further used to gain insight into the copper environment in both the Cu(II) and Cu(I) redox states. The experimental data were successfully rationalized by DFT models, offering valuable information on the electronic structure and coordination geometry of the copper center. Finally, the Cu(II)/Cu(I) redox potential was determined using two different methods at ca. 350 mV vs NHE and rationalized by DFT calculations. This integrated approach not only advances our knowledge of the active site properties of SmAA10 but also establishes a robust framework for future studies of similar enzymatic systems.

Diamond Keywords: Enzymes

Subject Areas: Chemistry, Biology and Bio-materials


Instruments: I20-Scanning-X-ray spectroscopy (XAS/XES)

Other Facilities: ID26 at ERSF

Added On: 31/05/2024 14:38

Discipline Tags:

Physical Chemistry Biochemistry Chemistry Inorganic Chemistry Life Sciences & Biotech

Technical Tags:

Spectroscopy X-ray Absorption Spectroscopy (XAS)