Functional variation among LPMOs revealed by the inhibitory effects of cyanide and buffer ions.

Enzymes known as lytic polysaccharide monooxygenases (LPMOs) are mono-copper polysaccharide-degrading peroxygenases that engage in several on- and off-pathway redox reactions involving O₂ and H₂O₂. Herein, we show that the known metalloenzyme inhibitor cyanide inhibits reductive activation of LPMOs by binding to the LPMO-Cu(II) state and that the degree of inhibition depends on the concentrations of the polysaccharide substrate, the reductant and H₂O₂. Importantly, this analysis revealed differences between fungal NcAA9C and bacterial SmAA10A, which have different secondary copper coordination spheres. These differences were also highlighted by the observation that phosphate, a commonly used buffer ion, strongly inhibits NcAA9C while not affecting reactions with SmAA10A. The results provide insight into LPMO inhibition and catalysis and highlight pitfalls in the analysis thereof.