Novel catalytic potential of a hyperthermostable mono‑copper oxidase (LPMO-AOAA17) for the oxidation of lignin monomers and depolymerisation of lignin dimer in aqueous media.

Lytic polysaccharide monooxygenase (LPMO) are mono‑copper enzymes known for the oxidative cleavage of recalcitrant polysaccharides with their intriguing and unique catalytic chemistry. Such impeccable oxidation potential has made them highly valuable in the enzymatic consortia for the degradation of ligno-cellulosic biomass. Bioinformatic analysis has revealed an unannotated LPMO gene in the genome of A. oryzae. Multiple sequence alignment showed the presence of conserved "histidine brace" of LPMO in the amino acid sequence of the enzyme. The enzyme, named as LPMO-AOAA17 was recombinantly expressed in E. coli BL21 and characterised for its substrate specificity. Recombinant enzyme did not show any characteristic cleavage of polysaccharides. However, it was found to be oxidising broad range of phenolic and non-phenolic monomers of lignin. Biochemical study revealed the optimum activity of LPMO-AOAA17 at pH 7 and was highly stable and active at 100 °C. The enzyme LPMO-AOAA17 was also observed to be stable after autoclaving at 121 °C and 15 psi. Thermal stability of the LPMO-AOAA17 was further confirmed through differential scanning calorimetry. GC-MS analysis has confirmed the catalysis of LPMO-AOAA17 for the depolymerisation of lignin dimer, guaicyl glycerol β-guaicyl ether into guaiacol. This study has first time documented the identification of a hyperthermostable LPMO for oxidative cleavage of β-O-4 linkage of lignin compounds to form aromatic products in aqueous media.