The effects of mixing on the enzymatic hydrolysis of lignocellulosic biomass

Biorefining of lignocellulosic biomass into biofuels and chemicals can help replace fossil resources and decrease anthropogenic greenhouse gas emissions. This thesis is focused on the effects of mixing on the enzymatic hydrolysis of pretreated biomass. Two different types of biomass were studied: softwood (Norway spruce and Scots pine), and the energy grass giant reed. Before enzymatic hydrolysis, the biomass was pretreated by either steam or sulfite pretreatment. The first part of the work concerns the connection between particle morphology and rheology of pretreated biomass, how such properties change during the course of enzymatic hydrolysis, and how the changes are influenced by reactor mixing. The second part examines the effects of mixing in stirred tank reactors on the enzymatic hydrolysis of different pretreated materials, and also attempts to explain the mechanisms behind the observed phenomena.The particle size reduction during enzymatic hydrolysis of steam pretreated spruce was primarily driven by reactor agitation. In the case of steam pretreated giant reed the particle size was mainly reduced by enzymatic hydrolysis. The rapid reduction in particle size of giant reed coincided with a rapid liquefaction. For steam pretreated softwood, the viscosity in fact increased at the beginning of enzymatic hydrolysis, followed by a gradual decrease during the remainder of the hydrolysis. This interesting phenomenon was in part linked to the type of pretreatment used on the softwood biomass. In contrast to steam pretreated softwood, the viscosity of sulfite pretreated spruce decreased rapidly during enzymatic hydrolysis. Efficient viscosity reduction in sulfite pretreated spruce was also achieved with very low doses of pure endoglucanase enzymes (0.1 mg protein per g glucan) without significant glucose release.The effect of mixing on the enzymatic hydrolysis was in part determined by the viscosity of the pretreated biomass. For steam pretreated spruce at low solid loading, decreasing the agitation rate had little effect on the the enzymatic hydrolysis. However, if the viscosity was increased by the addition of a thickening agent, the effect of agitation was much larger. For a substrate that underwent rapid initial viscosity reduction, such as steam pretreated giant reed, the enzymatic hydrolysis was almost independent of agitation rate. Another important factor determining the effect of mixing on the enzymatic hydrolysis was the level of product inhibition. If the glucose and cellobiose concentrations were high, as during high solid hydrolysis of steam pretreated spruce, low agitation rate had a large negative effect on the enzymatic hydrolysis. However, if the product concentration was kept low, as during SSF, the effect of agitation was much weaker. Overall, the results indicate that the decrease in hydrolysis rate occurred due to increased local product inhibition, caused by mass transfer limitations in the stagnant zones, formed in the reactor volume when under low intensity mixing. The rate of enzymatic hydrolysis appeared to be determined by flow regime, i.e. Reynolds number, rather than specific mixing power input. This implies that the negative effects of low agitation rate will be less of a problem in larger reactors

Effects of agitation on particle-size distribution and enzymatic hydrolysis of pretreated spruce and giant reed.

Mixing is an energy demanding process which has been previously shown to affect enzymatic hydrolysis. Concentrated biomass slurries are associated with high and non-Newtonian viscosities and mixing in these systems is a complex task. Poor mixing can lead to mass and/or heat transfer problems as well as inhomogeneous enzyme distribution, both of which can cause possible yield reduction. Furthermore the stirring energy dissipation may impact the particle size which in turn may affect the enzymatic hydrolysis. The objective of the current work was to specifically quantify the effects of mixing on particle-size distribution (PSD) and relate this to changes in the enzymatic hydrolysis. Two rather different materials were investigated, namely pretreated Norway spruce and giant reed

Scale-up of high-solid enzymatic hydrolysis of steam-pretreated softwood : the effects of reactor flow conditions

The importance of flow conditions during scale-up of high-solid enzymatic hydrolysis of steam-pretreated spruce was demonstrated by comparing hydrolysis rates between laboratory (2 L) and demonstration (4 m3) scale. A positive effect of increased agitation speed on the rate of enzymatic hydrolysis was found regardless of scale. Importantly, the hydrolysis rate was higher at the larger scale when compared at similar specific power inputs. Changes in the rheological properties of the pretreated material during the hydrolysis were followed by off-line measurements of apparent viscosity. This information was used to estimate the flow conditions in the reactors, i.e., average Reynolds numbers, which together with measured mixing power consumptions enabled a more detailed comparison between the scales. The hydrolysis yields correlated better with average Reynolds numbers than specific power input over the different scales. This indicates that mass transport limitations, caused by insufficient bulk flow, likely play a decisive role in determining the rate of enzymatic hydrolysis

Demonstration-scale enzymatic saccharification of sulfite-pulped spruce with addition of hydrogen peroxide for LPMO activation

The saccharification of lignocellulosic materials like Norway spruce is challenging due to the recalcitrant nature of the biomass, and it requires optimized and efficient pretreatment and enzymatic hydrolysis processes to make it industrially feasible. In this study, we report successful enzymatic saccharification of sulfite-pulped spruce (Borregaard's BALI™ process) at demonstration scale, achieved through the controlled delivery of hydrogen peroxide (H2O2) for the activation of lytic polysaccharide monooxygenases (LPMOs) present in the cellulolytic enzyme preparation. We achieved 85% saccharification yield in 4 days using industrially relevant conditions – that is, an enzyme dose of 4% (w/w dry matter of substrate) of the commercial cellulase cocktail Cellic CTec3 and a substrate loading of 12% (w/w). Addition of H2O2 and the resulting controlled and high LPMO activity had a positive effect on the rate of saccharification and the final sugar titer. Clearly, the high LPMO activity was dependent on feeding the reactors with the LPMO co-substrate H2O2, as in situ generation of H2O2 from molecular oxygen was limited. These demonstration-scale experiments provide a solid basis for the use of H2O2 to improve enzymatic saccharification of lignocellulosic biomass at large industrial scale

Mapping the potential for decentralized energy generation based on RES in Western Balkans

Although the countries of the Western Balkans are mostly electrified, there are still regions which do not have access to the electricity network or where the network capacity is insufficient. For the most part such areas are under special care of the state (i.e. underdeveloped, devastated by war, depopulated), on islands or in mountainous regions. Since the decentralized energy generation covers a broad range of technologies, including many renewable energy technologies that provide small-scale power at sites close to the users, such concept could be of interest for these locations. This paper identfies the areas in Western Balkans where such systems could be applied Consideration is given to geographical locations as well as possible applications. Wind hydro, solar photovoltaic, and biomass conversion systems were taken into consideration. Since the renewable energy sources data for Western Balkans region are rather scarce, the intention was to give a survey of the present situation and an estimate of future polential for decentralized energy generation based on renewable energy sources. The decentralized energy generation based on renewable energy sources in Western Balkans will find its niche easier for the users that will produce electricity for their own needs and for the users located in remote rural areas (off-grid applications)