Ethanol from lignocellulose : Management of by-products of hydrolysis

Abstract: Fuel ethanol can be produced from lignocellulosic materials, such as residues from agriculture and forestry. The polysaccharides of lignocellulose are converted to sugars by hydrolysis and the sugars can then be fermented to ethanol using microorganisms. However, during hydrolysis a wide range of by-products are also generated. By-product formation can affect ethanol yield and productivity. Management of by-products of hydrolysis is therefore important in the development of commercially viable production of cellulosic ethanol. Detoxification of inhibitory dilute-acid lignocellulose hydrolysates by treatment with Ca(OH)2 (overliming) efficiently improves the fermentability, but is associated with drawbacks like sugar degradation and CaSO4 precipitation. In factorial designed experiments, in which pH and temperature were varied, dilute-acid spruce hydrolysates were treated with Ca(OH)2, NH4OH or NaOH. The concentrations of sugars and inhibitory compounds were measured before and after the treatments. The fermentability was examined using the yeast Saccharomyces cerevisiae and compared with reference fermentations of synthetic medium without inhibitors. The treatment conditions were evaluated by comparing the balanced ethanol yield, which takes both the degradation of sugars and the ethanol production into account. Treatment conditions resulting in excellent fermentability and minimal sugar degradation were possible to find regardless of whether Ca(OH)2, NH4OH or NaOH was used. Balanced ethanol yields higher than those of the reference fermentations were achieved for hydrolysates treated with all three types of alkali. As expected, treatment with Ca(OH)2  gave rise to precipitated CaSO4. The NH4OH treatments gave rise to a brownish precipitate but the amounts of precipitate formed were relatively small. No precipitate was observed in treatments with NaOH. The findings presented can be used to improve the effectiveness of alkali detoxification of lignocellulose hydrolysates and to minimize problems with sugar degradation and formation of precipitates. Overexpression of different S. cerevisiae genes was investigated with the aim to engineer a biocatalyst with increased inhibitor tolerance. Overexpression of YAP1, a gene encoding a transcription factor, conveyed increased resistance to lignocellulose-derived inhibitors as well as to a dilute-acid hydrolysate of spruce. Recombinant Aspergillus niger expressing the Hypocrea jecorina endoglucanase Cel7B was cultivated on spent lignocellulose hydrolysate (stillage). The fungus simultaneously removed inhibitors present in the stillage and produced higher amounts of endoglucanase than when it was grown in a standard medium with comparable monosaccharide content. The concept can be applied for on-site production of enzymes in a cellulose-to-ethanol process and facilitate recycling of the stillage stream.