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The influence of enzymatic liquefaction and slurry rheology on high-solids cellulose hydrolysis van der Zwan, Timo William

Abstract

Enzyme-mediated hydrolysis of lignocellulosic materials for their conversion into bioproducts would benefit significantly if high solids concentrations (low water-to-biomass ratios) could be processed effectively. However, the fibrous nature of lignocellulosic biomass makes effective reaction mixing difficult, resulting in mass transfer limitations that reduce process yields. Overcoming these rheological challenges will require a better understanding of the substrate properties that influence slurry rheology, and in particular, how the action of carbohydrate-active enzymes can better facilitate slurry viscosity reduction, or ‘enzymatic liquefaction’. To this end, the work described in this thesis assessed: the underlying causes of the rheological challenge of high-solids bioconversion; the possible mechanisms of enzymatic liquefaction; how liquefaction is influenced by the nature of the substrate; and the roles the various enzymes play in liquefaction. It was apparent that the relationship between substrate properties, slurry rheology and high-solids hydrolysis kinetics is complex and multifaceted. Substrate–water interactions were shown to be a key determinant that influenced the mass transfer boundary and the scaling of yield stress with solids loading. The yield stress profiles of the various pretreated substrates varied extensively, indicating that ‘high solids’ is a relative, substrate-dependent quality. Substrate rheological characteristics, especially slurry yield stress, were shown to be directly linked to liquefaction efficiency and to reductions in hydrolysis yield with increasing solids loading. It appeared that enzyme-mediated liquefaction of biomass was achieved through a combination of material dilution, particle fragmentation and alteration of interparticle interactions. Cellobiohydrolases and endoglucanases were shown to be the key enzymes involved in these mechanisms. However, the effectiveness of the various enzymes was strongly influenced by the substrate’s physicochemical properties and concentration. Notably, an enzyme’s low-solids slurry viscosity-reducing capacity did not necessarily reflect its capacity to catalyze liquefaction at high solids loadings. Furthermore, reaction efficiencies tested at low solids loadings did not reliably predict efficiency at high solids due to substrate-specific rheological differences. In summary, this work provided rheological and enzymological insights into the highly disparate reaction kinetics and rheological challenge prevailing at commercially relevant substrate concentrations.

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