Depolymerization of Lignocellulose by Lytic Polysaccharide MonoOxygenases

Sammanfattning: Lignocellulose biomass is considered as one of the most potential and sustainable sources for the production of value-added chemicals and fuels while replacing the traditional petroleum resources. In a biorefinery, by employing biochemical conversion processes,cellulose present in the biomass is broken down into monomeric sugars which can belater converted into fuels or chemicals. This process is done with the help of different cellulose digesting enzymes (cellulases), isolated from natural cellulolytic organisms suchas saprophytic fungi.Lytic polysaccharide monooxygenases (LPMOs) are considered as one of the vital classesof enzymes in the bio-conversion of lignocellulose. They are copper active enzymes present naturally in cellulose degrading fungi. Unlike the traditional cellulases, they havea unique way of breaking cellulose using molecular oxygen or hydrogen peroxide as cosubstratein the presence of a reducing agent. Their ability to enhance the action of other cellulases in depolymerizing the cellulose, make them an integral part of today’s commercial cellulase cocktails.This thesis comprises the study about the action of lytic polysaccharide monooxygenaseson various cellulose substrates, both model and natural. The first part of the thesis focuses on the ability of an LPMO (MtLPMO9) and a traditional cellulase (MtEG5A), to act insynergism. The evaluation was done based on the release of oxidized and non-oxidized sugars and also on the ability to liquefy the substrates. It was observed that together, these two enzymes resulted in enhanced release of oxidized and non-oxidized sugars. Both were able to reduce viscosity of the substrates but no further synergistic effect was observed when added together.The second part focuses on the ability of LPMOs to accept electrons from lignins for their action of breaking cellulose chains. Three LPMOs, MtLPMO9, PcLPMO9D and NcLPMO9C, lignins from agricultural and forest biomass pretreated by various pretreatment methods were selected. It was demonstrated that lignins, both in isolatedand substrate bound form were able to act indirectly as reducing agents, by releasingsoluble low-molecular-weight molecules that act as mediators between enzyme and bulklignins. The structural and compositional properties of lignins also affected their ability toact as electron donors. In addition, the effect of biomass pretreatment methods on the lignin properties was also studied. The lignins from acid catalyzed organosolv pretreatment were found as the best candidates in supplying electrons to the enzymes.Interestingly, NcLPMO9C was not able to utilize lignins as electron donors requiring further investigation on their mechanism both in vivo and in vitro.