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Background: 2-Naphthol, a carbocation scavenger, is known to mitigate lignin condensation during the acidic processing of lignocellulosic biomass, which may benefit downstream processing of the resulting materials. Consequently, various raw materials have demonstrated improved enzymatic saccharification yields for substrates pretreated through autohydrolysis and dilute acid hydrolysis in the presence of 2-naphthol. However, 2-naphthol is toxic to ethanol-producing organisms, which may hinder its potential application. Little is known about the implications of 2-naphthol in combination with the pretreatment of softwood bark during continuous steam explosion in an industrially scalable system.

Results: The 2-naphthol-pretreated softwood bark was examined through spectroscopic techniques and subjected to separate hydrolysis and fermentation along with a reference excluding the scavenger and a detoxified sample washed with ethanol. The extractions of the pretreated materials with water resulted in a lower aromatic content in the extracts and stronger FTIR signals, possibly related to guaiacyl lignin, in the nonextractable residue when 2-naphthol was used during pretreatment. In addition, cyclohexane/acetone (9:1) extraction revealed the presence of pristine 2-naphthol in the extracts and increased aromatic content of the nonextractable residue detectable by NMR for the scavenger-pretreated materials. Whole-slurry enzymatic saccharification at 12% solids loading revealed that elevated saccharification recoveries after 48 h could not be achieved with the help of the scavenger. Glucose concentrations of 16.9 (reference) and 15.8 g/l (2-naphthol) could be obtained after 48 h of hydrolysis. However, increased inhibition during fermentation of the scavenger-pretreated hydrolysate, indicated by yeast cell growth, was slight and could be entirely overcome by the detoxification stage. The ethanol yields from fermentable sugars after 24 h were 0.45 (reference), 0.45 (2-naphthol), and 0.49 g/g (2-naphthol, detoxified).

Conclusion: The carbocation scavenger 2-naphthol did not increase the saccharification yield of softwood bark pretreated in an industrially scalable system for continuous steam explosion. On the other hand, it was shown that the scavenger's inhibitory effects on fermenting microorganisms can be overcome by controlling the pretreatment conditions to avoid cross-inhibition or detoxifying the substrates through ethanol washing. This study underlines the need to jointly optimize all the main processing steps.

Biomass from four different Nordic microalgal species, grown in BG-11 medium or synthetic wastewater (SWW), was explored as inexpensive carbohydrate-rich feedstock for polyhydroxybutyrate (PHB) production via microbial fermentation. Thermochemical pre-treatment (acid treatment followed by autoclavation) with 2% hydrochloric acid or 1% sulphuric acid (v/v) was used to maximize sugar yield prior to fermentation. Pre-treatment resulted in ∼5-fold higher sugar yield compared to the control. The sugar-rich hydrolysate was used as carbon source for the PHB-producing extremophilic bacterium Halomonas halophila. Maximal PHB production was achieved with hydrolysate of Chlorococcum sp. (MC-1) grown on BG-11 medium (0.27 ± 0.05 g PHB/ g DW), followed by hydrolysate derived from Desmodesmus sp. (RUC-2) grown on SWW (0.24 ± 0.05 g PHB/ g DW). Nordic microalgal biomass grown on wastewater therefore can be used as cheap feedstock for sustainable bioplastic production. This research highlights the potential of Nordic microalgae to develop a biobased economy.

Spent mushroom substrates (SMS) from cultivation of shiitake (Lentinula edodes) on three hardwood species were investigated regarding their potential for cellulose saccharification and for ethanolic fermentation of the produced hydrolysates. High glucan digestibility was achieved during enzymatic saccharification of the SMSs, which was related to the low mass fractions of lignin and xylan, and it was neither affected by the relative content of lignin guaiacyl units nor the substrate crystallinity. The high nitrogen content in SMS hydrolysates, which was a consequence of the fungal pretreatment, was positive for the fermentation, and it ensured ethanol yields corresponding to 84–87% of the theoretical value in fermentations without nutrient supplementation. Phenolic compounds and acetic acid were detected in the SMS hydrolysates, but due to their low concentrations, the inhibitory effect was limited. The solid leftovers resulting from SMS hydrolysis and the fermentation residues were quantified and characterized for further valorisation.

Microalgae are promising candidates for sustainable wastewater treatment coupled to the production of biofuel, bioplastic and/or bio-fertilizers. In Nordic countries, however, light is a limiting factor for photosynthesis and biomass production during the winter season. Compared to municipal wastewater, industrial wastewater streams from the pulp and paper industry contain lower amounts of nitrogen, but high concentrations of carbon sources, which could be utilized by microalgae to enhance biomass production in limiting light. This study focused on the utilization of methanol, glycerol and xylose by five different Nordic microalgae [Chlorella vulgaris (13–1), Coelastrella sp. (3–4), Desmodesmus sp. (2–6), Chlorococcum sp. (MC1) and Scotiellopsis reticulata (UFA-2)] grown under mixotrophic conditions. Two of these strains, i.e., Chlorococcum sp. (MC1) and Scotiellopsis reticulata (UFA-2) were able to grow in the presence of xylose or methanol at concentrations of 6 g L^–1, or 3%, respectively, in a 12/12 h day/night cycle. HPLC analysis confirmed the consumption of those substrates. Glycerol (2.3 g L^–1) was tolerated by all strains and increased growth for Chlorella vulgaris (13–1), while higher concentrations (20 g L^–1) were only tolerated by Chlorococcum sp. (MC-1). Fourier-transform infrared spectroscopy, performed after growth in presence of the dedicated carbon source, indicated an increase in the fingerprint region of the carbohydrate fraction. This was particularly the case for Chlorococcum sp. (MC1), when grown in presence of glycerol, and Scotiellopsis reticulata (UFA-2), when grown in presence of xylose. Therefore, these strains could be potential candidates for the production of biofuels, e.g., bioethanol or biogas. We could show that Nordic microalgae are able to grow on various carbon sources; the actual uptake rates are low during a 12/12 h day/night cycle requesting additional optimization of the cultivation conditions. Nonetheless, their potential to use pulp and paper waste-streams for cheap and sustainable biomass production is high and will support the development of new technologies, turning waste-streams into resources in a circular economy concept.

Hydrolysis lignin, i.e., the hydrolysis residue of cellulosic ethanol plants, was extracted with the green solvent γ-valerolactone (GVL). Treatments at 170–210 °C were performed with either non-acidified GVL/water mixtures (NA-GVL) or with mixtures containing sulfuric acid (SA-GVL). SA-GVL treatment at 210 °C resulted in the highest lignin solubilization (64% (w/w) of initial content), and 76% of the solubilized mass was regenerated by water-induced precipitation. Regenerated lignins were characterized through compositional analysis with sulfuric acid, as well as using pyrolysis–gas chromatography/mass spectrometry (Py-GC/MS), high-performance size-exclusion chromatography (HPSEC), solid-state cross-polarization/magic angle spinning ¹³C nuclear magnetic resonance (CP/MAS ¹³C NMR) spectroscopy, ¹H^–13C heteronuclear single-quantum coherence NMR (HSQC NMR), and Fourier-transform infrared (FTIR) spectroscopy. The characterization revealed that the main difference between regenerated lignins was their molecular weight. Molecular weight averages increased with treatment temperature, and they were higher and had broader distribution for SA-GVL lignins than for NA-GVL lignins.

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.

Bagasse, a major by-product of sugarcane-processing industries, has potential to play a significant role as feedstock for production of cellulosic ethanol, platform chemicals, and bio-based commodities. Pretreatment is essential for efficient processing of lignocellulosic feedstocks by biochemical conversion. In this work, auto catalyzed (A-HTP) and dilute sulfuric-acid-catalyzed (SA-HTP) hydrothermal pretreatment of sugarcane bagasse was investigated, setting the temperature (175-205 degrees C) and the time (4-51 min) in such a way that the severity factor (SF) was always maintained at one of three predetermined values (2.8, 3.8, and 4.8). The investigation covered the effects of different operational pretreatment conditions on (i) the formation of sugars and water-soluble bioconversion inhibitors, including newly discovered inhibitors such as formaldehyde and pbenzoquinone, in the pretreatment liquid, (ii) the chemical composition and recovery of constituents in the solid phase, as determined using two-step treatment with sulfuric acid, Py-GC/MS, and solid-state NMR, (iii) pseudo lignin formation, (iv) furan aldehydes in condensates from the gas phase, (v) enzymatic digestibility of pretreated solids, (vi) enzyme inhibition by pretreatment liquids, and (vii) fermentability of pretreatment liquids using Saccharomyces cerevisiae yeast. Glucose and xylose were the predominant sugars in pretreatment liquids from SAHTP and A-HTP, respectively. For A-HTP, the enzymatic digestibility of the pretreated solids was proportional to the SF, while for SA-HTP no clear trend was observed. The best enzymatic digestibility (above 80%) was achieved for A-HTP performed at SF 4.8. The highest total yields of glucose and xylose, the predominant sugars, were achieved for A-HTP at SF 3.8 and temperatures of 190 degrees C and 205 degrees C. The fermentability of the pretreatment liquids by Saccharomyces cerevisiae was lower for SA-HTP than for A-HTP. The investigation suggests that hydrothermal pretreatment of sugarcane bagasse can be performed with good results without addition of sulfuric acid, but that the conditions must be just harsh enough to almost quantitatively solubilize the hemicelluloses.

The effects of the redox environment on acidic hydrothermal pretreatment were investigated in experiments with sugarcane bagasse (190 degrees C, 14 min) and Norway spruce (205 degrees C, 5 min). To modulate the redox environment, pretreatment was performed without gas addition, with N-2 , or with O-2. Analyses covered pretreated solids, pretreatment liquids, condensates, enzymatic digestibility, and inhibitory effects of pretreatment liquids on yeast. Addition of gas, especially O-2 , resulted in increased severity, as reflected by up to 18 percent units lower recoveries of pretreated solids, up to 31 percent units lower glucan recoveries, improved hemicellulose removal, formation of pseudo-lignin, improved overall glucan conversion, and increased concentrations of several microbial inhibitors. Some inhibitors, such as formaldehyde and coniferyl aldehyde, did not, however, follow that pattern. TAC (Total Aromatic Content) values reflected inhibitory effects of pretreatment liquids. This study demonstrates how gas addition can be used to modulate the severity of acidic hydrothermal pretreatment.

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.

Cassava stems are an abundant feedstock that is becoming attractive for biochemical conversion to fuels and chemicals. Since cassava stems are rich in both cellulose and starch, carefully designed pretreatment and digestion procedures are required for achieving high glucan recovery. In this study, partially de-starched cassava stems resulting from a water extraction stage were hydrolyzed with amylases, and the resulting starch-depleted material was pretreated with dilute sulfuric acid, and submitted to enzymatic hydrolysis of cellulose. The effects of acid pretreatment on glucan recovery, enzymatic convertibility, and by-product formation were investigated using a Box-Behnken experimental design with temperature (165-195 degrees C), time (5-35 min), and acid concentration (0.2-1.0%) as independent variables. In further experimental series, the time period was extended up to 110 min while maintaining temperature at 195 degrees C and sulfuric acid concentration at 0.6%. Using those conditions, pretreatment for 50 min gave the best results (83.8% enzymatic convertibility of pretreated cellulose, and (similar to)72% overall glucan-to-glucose conversion).