PRODUCTION OF THE LYTIC POLYSACCHARIDE MONOOXYGENASE TrCel61A FROM Trichoderma reesei BY Komagataella phaffii IN BIOREACTOR

Cellulose is the most abundant natural biopolymer on the earth. However, its recalcitrance is one of the main limiting factors for the deconstruction of this polysaccharide in monomers. Hydrolytic enzymes as cellulases and hemicellulases have been used for glucose production and subsequent alcoholic fermentation. In addition, oxidative enzymes have been used to increase the release of glucose together with the hydrolases. Lytic polysaccharide monooxygenases (LPMOs) present an oxidative activity and are central component of commercial enzyme cocktails used for the industrial production of lignocellulosic ethanol. The LPMO TrCel61A from Trichoderma reesei was already produced by different hosts including Komagataella phaffii in bioreactor. In this work, we investigated the recombinant production of this enzyme by K. phaffii cells using methanol feed and pO2 fixed at 15% and 30%. The results obtained in the cultures A (pO2 15%) and B (pO2 30%) were 37.9 and 33.0 g/L for Cell biomass and 0.23 and 0.33 g/L to extracellular protein. Molecular mass calculated from the SDS PAGE analysis showed a protein with approximately 60 kDa. LPMO produced in fixed pO2 at 30% and methanol feed of 3 g/L per hour is promising approach to obtain this protein at high levels.