Search

The role of endocellulases and endoxylanase during liquefaction and saccharification of hydrothermally pretreated wheat straw was studied. The use of a flow-loop setup with in-line magnetic resonance imaging enabled frequent measurements of viscosity at 55°C during saccharification at 6% total solids content. Viscosity data were complemented with off-line measurements of fiber lengths and release of soluble sugars. A clear correlation between fiber attrition and a decrease in viscosity was found. Fiber lengths and viscosity dropped quickly within the first hour and then stagnated, while sugar yields increased substantially thereafter, illustrating that liquefaction and saccharification are separate mechanisms. Both endoglucanase and endoxylanase were shown to have a significant effect on viscosity during liquefaction while the addition of endoxylanase also increased sugar yield.

Palm kernel press cake (PKC) is a residue of palm oil extraction, which was found to contain 48.5% of total carbohydrates of which 35.2% was mannan. The present study examines enzymatic hydrolysis of polysaccharides from the cell-wall material present in PKC to obtain monosaccharides that can be substrate in various fermentation processes such as ethanol production. The requirements for pretreatment were investigated and it was found that mannan in PKC was readily hydrolysed without any pretreatment. Several enzyme preparations were tested and Mannaway 25L was found as the best for releasing mannose, and Gammanase 1.0L worked well in degrading cellulose and mannose. Binary mixtures of enzymes were tested to increase the conversion, and 1:1 mixture of Mannaway 25L and Gammanase 1.0L showed good synergistic effect releasing 30% more mannose than the sum obtained using these enzymes individually. Using an enzyme loading of 2.3 mg protein/g PKC resulted in 63% of mannan in PKC being hydrolysed to mannose in 24 h, and in 96 h a total of 365 g mannose and glucose could be produced per kg PKC. Finally, PKC was hydrolysed and fermented using Saccharomyces cerevisiae with an ethanol yield of 125 g/kg PKC.

Recycling of enzymes has a potential interest during cellulosic bioethanol production as purchasing enzymes is one of the largest expenses in the process. By recycling enzymes after distillation, loss of sugars and ethanol are avoided, but depending on the distillation temperature, there is a potential risk of enzyme degradation. Studies of the rate of enzyme denaturation based on estimation of the denaturation constant K D was performed using a novel distillation setup allowing stripping of ethanol at 50–65 °C. Experiments were performed in a pilot-scale stripper, where the effect of temperature (55–65 °C) and exposure to gas–liquid and liquid–heat transmission interfaces were tested on a mesophilic and thermostable enzyme mixture in fiber beer and buffer. Lab-scale tests were included in addition to the pilot-scale experiments to study the effect of shear, ethanol concentration, and PEG on enzyme stability. When increasing the temperature (up to 65 °C) or ethanol content (up to 7.5 % w/v), the denaturation rate of the enzymes increased. Enzyme denaturation occurred slower when the experiments were performed in fiber beer compared to buffer only, which could be due to PEG or other stabilizing substances in fiber beer. However, at extreme conditions with high temperature (65 °C) and ethanol content (7.5 % w/v), PEG had no enzyme stabilizing effect. The novel distillation setup proved to be useful for maintaining enzyme activity during ethanol extraction.