Ethanol from lignocellulose : Alkali detoxification of dilute-acid spruce hydrolysates

Detoxification of dilute-acid lignocellulose hydrolysates by treatment with Ca(OH)2 (overliming) efficiently improves the production of fuel ethanol, 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 possibility that the ammonium ions from the NH4OH treatments gave a positive effect as an extra source of nitrogen during the fermentations was excluded after experiments in which NH4Cl was added to the medium. 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

Methods for improvement of enzymatic hydrolysis of lignocellullosic material

The present invention relates to a method of enzymatic hydrolysis of a lignocellulosic material, comprising the steps of: a) pretreating the lignocellulosic material to obtain a slurry having a pH of less than 6; b) adding NaOH, Ca(OH)2 and/or CaO to the slurry to increase its pH to at least 8, said addition being carried out at a slurry temperature of at least 60 °C; c) reducing the pH of the slurry to below 7; and optionally cooling the slurry from step b) to a temperature below 60 °C; and d) adding hydrolytic enzymes to the slurry from c) and allowing the slurry to hydrolyze wherein no washing of the slurry is performed prior to step d)WO 2013/000696 A1, PCT/EP2012/061021</p

Conditioning of pretreated birch by liquid-liquid organic extractions to improve yeast fermentability and enzymatic digestibility

By-products from hydrothermal pretreatment of lignocellulosic biomass inhibit enzymatic saccharification and microbial fermentation. Three long-chain organic extractants (Alamine 336, Aliquat 336 and Cyanex 921) were compared to two conventional organic solvents (ethyl acetate and xylene) with regard to conditioning of birch wood pretreatment liquid (BWPL) for improved fermentation and saccharification. In the fermentation experiments, extraction with Cyanex 921 resulted in the best ethanol yield, 0.34 ± 0.02 g g−1 on initial fermentable sugars. Extraction with xylene also resulted in a relatively high yield, 0.29 ± 0.02 g g−1, while cultures consisting of untreated BWPL and BWPL treated with the other extractants exhibited no ethanol formation. Aliquat 336 was most efficient with regard to removing by-products, but the residual Aliquat after the extraction was toxic to yeast cells. Enzymatic digestibility increased by 19-33% after extraction with the long-chain organic extractants. The investigation demonstrates that conditioning with long-chain organic extractants has the potential to relieve inhibition of both enzymes and microbes

Bioconversion of lignocellulose : inhibitors and detoxification

Bioconversion of lignocellulose by microbial fermentation is typically preceded by an acidic thermochemical pretreatment step designed to facilitate enzymatic hydrolysis of cellulose. Substances formed during the pretreatment of the lignocellulosic feedstock inhibit enzymatic hydrolysis as well as microbial fermentation steps. This review focuses on inhibitors from lignocellulosic feedstocks and how conditioning of slurries and hydrolysates can be used to alleviate inhibition problems. Novel developments in the area include chemical in-situ detoxification by using reducing agents, and methods that improve the performance of both enzymatic and microbial biocatalysts

Factors affecting detoxification of softwood enzymatic hydrolysates using sodium dithionite

Conditioning of lignocellulosic hydrolysates with sulfur oxyanions, such as dithionite, is one of the most potent methods to improve the fermentability by counteracting effects of inhibitory by-products generated during hydrothermal pretreatment under acidic conditions. The effects of pH, treatment temperature, and dithionite dosage were explored in experiments with softwood hydrolysates, sodium dithionite, and Saccharomyces cerevisiae yeast. Treatments with dithionite at pH 5.5 or 8.5 gave similar results with regard to ethanol productivity and yield on initial glucose, and both were always at least ~20% higher than for treatment at pH 2.5. Experiments in the dithionite concentration range 5.0–12.5 mM and the temperature range 23–110◦ C indicated that treatment at around 75◦ C and using intermediate dithionite dosage was the best option (p ≤ 0.05). The investigation indicates that selection of the optimal temperature and dithionite dosage offers great benefits for the efficient fermentation of hydrolysates from lignin-rich biomass, such as softwood residues

Comparison of Efficiency and Cost of Methods for Conditioning of Slurries of Steam-Pretreated Softwood

Inhibitors formed during pretreatment impair lignocellulose bioconversion by making enzymatic saccharification and microbial fermentation less efficient, but conditioning of slurries and hydrolysates can improve fermentability and sometimes also enzymatic digestibility. Conditioning of pretreated softwood using four industrial reducing agents (sodium sulfite, sodium dithionite, sodium borohydride, and hydrogen) was compared with standard methods, such as overliming and treatment with activated charcoal. A dosage of approx. 1 mM sulfur oxyanion (sulfite or dithionite) per percent water-insoluble solids (WIS) in the slurry was found to result in good fermentability. Treatment of 10–20% WIS slurries with 15 mM sulfur oxyanion under mild reaction conditions (23°C, pH 5.5) resulted in sulfonation of the solid phase and saccharification improvements of 18–24% for dithionite and 13–16% for sulfite. Among the different conditioning methods studied, treatment of slurries with sodium sulfite was superior with respect to cost-efficient improvement of fermentability. Treatments of slurry or pretreatment liquid with 15 mM sulfite or dithionite resulted in 58–76% reduction of the content of formaldehyde. The comparison indicates that conditioning of pretreated biomass using sulfur oxyanions warrants further attention

Ozone detoxification of steam-pretreated Norway spruce

Background: Pretreatment of lignocellulose for biochemical conversion commonly results in formation of by-products that inhibit microorganisms and cellulolytic enzymes. To make bioconversion processes more efficient, inhibition problems can be alleviated through conditioning. Ozone is currently commercially employed in pulp and paper production for bleaching, as it offers the desirable capability to disrupt unsaturated bonds in lignin through an ionic reaction known as ozonolysis. Ozonolysis is more selective towards lignin than cellulose, for instance, when compared to other oxidative treatment methods, such as Fenton's reagent. Ozone may thus have desirable properties for conditioning of pretreated lignocellulose without concomitant degradation of cellulose or sugars. Ozone treatment of SO2- impregnated steam-pretreated Norway spruce was explored as a potential approach to decrease inhibition of yeast and cellulolytic enzymes. This novel approach was furthermore compared to some of the most effective methods for conditioning of pretreated lignocellulose, i.e., treatment with alkali and sodium dithionite. Results: Low dosages of ozone decreased the total contents of phenolics to about half of the initial value and improved the fermentability. Increasing ozone dosages led to almost proportional increase in the contents of total acids, including formic acid, which ultimately led to poor fermentability at higher ozone dosages. The decrease of the contents of furfural and 5-hydroxymethylfurfural was inversely proportional (R-2 &gt; 0.99) to the duration of the ozone treatment, but exhibited no connection with the fermentability. Ozone detoxification was compared with other detoxification methods and was superior to treatment with Fenton's reagent, which exhibited no positive effect on fermentability. However, ozone detoxification was less efficient than treatment with alkali or sodium dithionite. High ozone dosages decreased the inhibition of cellulolytic enzymes as the glucose yield was improved with 13 % compared to that of an untreated control. Conclusions: Low dosages of ozone were beneficial for the fermentation of steam-pretreated Norway spruce, while high dosages decreased the inhibition of cellulolytic enzymes by soluble components in the pretreatment liquid. While clearly of interest for conditioning of lignocellulosic hydrolysates, future challenges include finding conditions that provide beneficial effects both with regard to enzymatic saccharification and microbial fermentation

Bioconversion of waste fiber sludge to bacterial nanocellulose and use for reinforcement of CTMP paper sheets

Utilization of bacterial nanocellulose (BNC) for large-scale applications is restricted by low productivity in static cultures and by the high cost of the medium. Fiber sludge, a waste stream from pulp and paper mills, was enzymatically hydrolyzed to sugar, which was used for the production of BNC by the submerged cultivation of Komagataeibacter xylinus. Compared with a synthetic glucose-based medium, the productivity of purified BNC from the fiber sludge hydrolysate using shake-flasks was enhanced from 0.11 to 0.17 g/(L x d), although the average viscometric degree of polymerization (DPv) decreased from 6760 to 6050. The cultivation conditions used in stirred-tank reactors (STRs), including the stirring speed, the airflow, and the pH, were also investigated. Using STRs, the BNC productivity in fiber-sludge medium was increased to 0.32 g/(L x d) and the DPv was increased to 6650. BNC produced from the fiber sludge hydrolysate was used as an additive in papermaking based on the chemithermomechanical pulp (CTMP) of birch. The introduction of BNC resulted in a significant enhancement of the mechanical strength of the paper sheets. With 10% (w/w) BNC in the CTMP/BNC mixture, the tear resistance was enhanced by 140%. SEM images showed that the BNC cross-linked and covered the surface of the CTMP fibers, resulting in enhanced mechanical strength.Bio4Energ