Mechanism of hydrogen peroxide formation by lytic polysaccharide monooxygenase. Issue 2 (25th October 2018)

• Record Type: Journal Article
• Authors: Caldararu, Octav; Oksanen, Esko; Ryde, Ulf; Hedegård, Erik D.
• Is Part Of: Chemical science. Volume 10:Issue 2(2019)
• Page Start: 576
• Page End: 586
• Publication Date: 2018-10-25
• Subjects: Chemistry -- Periodicals; 540.5
• DOI: 10.1039/c8sc03980a
• Languages: English
• ISSNs: 2041-6520

Abstract

Abstract : A mechanism for the formation of hydrogen peroxide by lytic polysaccharide monooxygenases (LPMOs) in the absence of substrate is proposed. Abstract : Lytic polysaccharide monooxygenases (LPMOs) are copper-containing metalloenzymes that can cleave the glycosidic link in polysaccharides. This could become crucial for production of energy-efficient biofuels from recalcitrant polysaccharides. Although LPMOs are considered oxygenases, recent investigations have shown that H2 O2 can also act as a co-substrate for LPMOs. Intriguingly, LPMOs generate H2 O2 in the absence of a polysaccharide substrate. Here, we elucidate a new mechanism for H2 O2 generation starting from an AA10-LPMO crystal structure with an oxygen species bound, using QM/MM calculations. The reduction level and protonation state of this oxygen-bound intermediate has been unclear. However, this information is crucial to the mechanism. We therefore investigate the oxygen-bound intermediate with quantum refinement (crystallographic refinement enhanced with QM calculations), against both X-ray and neutron data. Quantum refinement calculations suggest a Cu(ii )–O−2 system in the active site of the AA10-LPMO and a neutral protonated –NH2 state for the terminal nitrogen atom, the latter in contrast to the original interpretation. Our QM/MM calculations show that H2 O2 generation is possible only from a Cu(i ) center and that the most favourable reaction pathway is to involve a nearby glutamate residue, adding two electrons and two protons to the Cu(ii )–O−2 system, followed by dissociation of H2 O2 .