Challenges and opportunities in mimicking non-enzymatic brown-rot decay mechanisms for pretreatment of Norway spruce

The recalcitrance bottleneck of lignocellulosic materials presents a major challenge for biorefineries, including second-generation biofuel production. Because of their abundance in the northern hemisphere, softwoods, such as Norway spruce, are of major interest as a potential feedstock for biorefineries. In nature, softwoods are primarily degraded by basidiomycetous fungi causing brown rot. These fungi employ a non-enzymatic oxidative system to depolymerize wood cell wall components prior to depolymerization by a limited set of hydrolytic and oxidative enzymes. Here, it is shown that Norway spruce pretreated with two species of brown-rot fungi yielded more than 250% increase in glucose release when treated with a commercial enzyme cocktail and that there is a good correlation between mass loss and the degree of digestibility. A series of experiments was performed aimed at mimicking the brown-rot pretreatment, using a modified version of the Fenton reaction. A small increase in digestibility after pretreatment was shown where the aim was to generate reactive oxygen species within the wood cell wall matrix. Further experiments were performed to assess the possibility of performing pretreatment and saccharification in a single system, and the results indicated the need for a complete separation of oxidative pretreatment and saccharification. A more severe pretreatment was also completed, which interestingly did not yield a more digestible material. It was concluded that a biomimicking approach to pretreatment of softwoods using brown-rot fungal mechanisms is possible, but that there are additional factors of the system that need to be known and optimized before serious advances can be made to compete with already existing pretreatment methods.