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  • 1.
    Martín, Carlos
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Wu, Guochao
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Wang, Zhao
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Stagge, Stefan
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Jönsson, Leif J
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Formation of microbial inhibitors in steam-explosion pretreatment of softwood impregnated with sulfuric acid and sulfur dioxide2018Ingår i: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 262, s. 242-250Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Wood chips of Norway spruce were pretreated by steam explosion at 195–215 °C after impregnation with either sulfuric acid (SA) or sulfur dioxide (SD). The effects of different pretreatment conditions on formation of microbial inhibitors were investigated, and the inhibitory effects on yeast of pretreatment liquids and of specific inhibitors that were found in the pretreatment liquids were elucidated. Whereas the concentrations of most inhibitors increased with increasing pretreatment temperatures, there were exceptions, such as formaldehyde and p-hydroxybenzaldehyde. The highest concentration of each inhibitor was typically found in SD-pretreated material, but formic acid was an exception. The toxic effects on yeast were studied using concentrations corresponding to loadings of 12 and 20% total solids (TS). Among individual inhibitors that were quantitated in pretreatment liquids, the concentrations of formaldehyde were by far most toxic. There was no or minimal yeast growth in the formaldehyde concentration range (5.8–7.7 mM) corresponding to 12% TS.

  • 2.
    Wang, Zhao
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Recalcitrance of wood to biochemical conversion: feedstock properties, pretreatment, saccharification, and fermentability2018Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
    Abstract [en]

    Lignocellulose is an inexpensive and abundant renewable resource that can be used to produce advanced biofuels, green chemicals, and other bio-based products. Pretreatment and efficient enzymatic saccharification are essential features of bioconversion of lignocellulosic biomass. The aims of the research were to achieve a better understanding of the recalcitrance of woody biomass to bioconversion, to explore different pretreatment techniques that can be used to decrease the recalcitrance of the biomass and improve the digestibility of the cellulose, and to investigate by-products of acid pretreatment that cause enzymes and microorganisms to work less efficiently.

    The recalcitrance of wood from aspen, birch, and spruce was investigated before and after acid pretreatment. Before pretreatment, birch exhibited the highest recalcitrance, which was attributed to structural factors. After pretreatment, spruce showed the highest recalcitrance, which was attributed to chemical factors, such as high lignin content. Deacetylation of hybrid aspen in planta by a CE5 acetyl xylan esterase decreased the recalcitrance, and the glucose yield of enzymatic saccharification of non-pretreated wood increased with 27%.

    Pretreatment options based on ionic liquids and steam explosion were further explored. The effects of the anionic constituents of a series of imidazolium-based ionic liquids on pretreatment of aspen and spruce were investigated. [HSO4]− was efficient only for aspen, which was attributed to acid degradation of xylan. [MeCO2]− was efficient for both aspen and spruce, which was attributed to its capability to create a disordered cell wall structure rather than to removal of lignin and hemicellulose. A comparison was made between using sulfuric acid and sulfur dioxide for pretreatment of spruce. Although sulfur dioxide resulted in a pretreatment liquid that was more inhibitory to both enzymes and yeast, it was still superior to pretreatment with sulfuric acid, a phenomenon that was attributed to the particle size of the pretreated material.

    In a comparison of microbial inhibitors in pretreatment liquids from steam explosion of spruce, formaldehyde was found to be the most important inhibitor of yeast. Enzyme inhibition by catalytically non-productive adsorption to lignins and pseudo-lignin was investigated using quantitative proteomics. The results indicate that protein adsorption to pseudo-lignin can be as extensive as adsorption to real lignin. 

  • 3.
    Wang, Zhao
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Gräsvik, John
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Jönsson, Leif J.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Winestrand, Sandra
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Comparison of [HSO4](-), [Cl](-) and [MeCO2](-) as anions in pretreatment of aspen and spruce with imidazolium-based ionic liquids2017Ingår i: BMC Biotechnology, ISSN 1472-6750, E-ISSN 1472-6750, Vol. 17, artikel-id 82Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Background: Ionic liquids (ILs) draw attention as green solvents for pretreatment of lignocellulose before enzymatic saccharification. Imidazolium-based ILs with different anionic constituents ([HSO4], [Cl], [MeCO2]) were compared with regard to pretreatment of wood from aspen and spruce. The objective was to elucidate how the choice of anionic constituent affected the suitability of using the IL for pretreatment of hardwood, such as aspen, and softwood, such as spruce. The investigation covered a thorough analysis of the mass balance of the IL pretreatments, the effects of pretreatment on the cell wall structure as assessed by fluorescence microscopy, and the effects of pretreatment on the susceptibility to enzymatic saccharification. Torrefied aspen and spruce were included in the comparison for assessing how shifting contents of hemicelluloses and Klason lignin affected the susceptibility of the wood to IL pretreatment and enzymatic saccharification.

    Results: The glucose yield after IL pretreatment increased in the order [Cl] < [HSO4] < [MeCO2] for aspen, but in the order [HSO4] < [Cl] < [MeCO2] for spruce. For both aspen and spruce, removal of hemicelluloses and lignin increased in the order [Cl] < [MeCO2] < [HSO4]. Fluorescence microscopy indicated increasingly disordered cell wall structure following the order [HSO4] < [Cl] < [MeCO2]. Torrefaction of aspen converted xylan to pseudo-lignin and changed the glucose yield order to [HSO4] < [Cl] < [MeCO2].

    Conclusions: The acidity of [HSO4] caused extensive hydrolysis of xylan, which facilitated pretreatment of xylan-rich hardwood. Apart from that, the degree of removal of hemicelluloses and lignin did not correspond well with the improvement of the enzymatic saccharification. Taken together, the saccharification results were found to mainly reflect (i) the different capacities of the ILs to disorder the cell wall structure, (ii) the recalcitrance caused by high xylan content, and (iii) the capacity of the [HSO4]-based IL to hydrolyze xylan.

  • 4.
    Wang, Zhao
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Jönsson, Leif J
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Comparison of catalytically non-productive adsorption of fungal proteins to lignins and pseudo-lignin using isobaric mass taggingManuskript (preprint) (Övrigt vetenskapligt)
  • 5.
    Wang, Zhao
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Jönsson, Leif J.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Comparison of catalytically non-productive adsorption of fungal proteins to lignins and pseudo-lignin using isobaric mass tagging2018Ingår i: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 268, s. 393-401Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Catalytically non-productive adsorption of fungal enzymes to pseudo-lignin (PL) was compared to adsorption to lignin preparations derived from different sources (SL, spruce; BL, birch; OL, beech) using different methods [steam pretreatment/enzymatic saccharification (SL, BL) and organosolv processing (OL)]. The protein adsorption to the SL was more extensive than the adsorption to the hardwood lignins, which was relatively similar to the adsorption to the PL. The adsorption patterns of 13 individual proteins were studied using isobaric mass tagging with TMTsixplex reagent and LC-MS/MS analysis. The results suggest that, on an average, adsorption of proteins equipped with carbohydrate-binding modules, such as the cellulases CBHI, EGII, and EGIV, was less dependent on the quality of the lignin/PL than adsorption of other proteins, such as beta-Xyl, Xyn-1, and Xyn-2, which are involved in xylan degradation.

  • 6.
    Wang, Zhao
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Pawar, PM
    Derba-Maceluch, Marta
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysiologisk botanik.
    Hedenström, Mattias
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Chong, Sun-Li
    Tenkanen, M
    Mellerowicz, E
    Jönsson, Leif J
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Bioprocessing properties of hybrid aspen expressing a Carbohydrate Esterase Family 5 acetyl xylan esterase under control of a wood-specific promoterManuskript (preprint) (Övrigt vetenskapligt)
  • 7.
    Wang, Zhao
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Winestrand, Sandra
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Gillgren, Thomas
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Jönsson, Leif J.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Chemical and structural factors influencing enzymatic saccharification of wood from aspen, birch and spruce2018Ingår i: Biomass and Bioenergy, ISSN 0961-9534, E-ISSN 1873-2909, Vol. 109, s. 125-134Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    The susceptibility of untreated and sulfuric-acid-pretreated aspen, birch, and spruce to analytical enzymatic saccharification was studied in relation to their chemical composition and physical-structural features. The analytical data collected covered the mass fractions of lignin, carbohydrates, and extractives, the release of acetic acid, formic acid, and uronic acids by acid and alkaline hydrolysis, crystallinity and crystallite size, syringyl: guaiacyl (S:G) ratio of lignin, cellulose accessibility, FTIR spectra, images from SEM and fluorescence microscopy, and susceptibility to enzymatic saccharification using enzyme mixtures with and without supplementary xylanase.In the absence of pretreatment the mass fraction yield of Glc on the original dry wood in the analytical enzymatic saccharification increased in the order birch (16 g kg−1) < spruce (35 g kg−1) < aspen (150 g kg−1). After acid pretreatment, the order changed to spruce (170 g kg−1) < aspen (290 g kg−1), birch (290 g kg−1). The relatively high recalcitrance of untreated birch was not possible to relate to mass fraction of lignin, S:G ratio, cellulose crystallinity, or mass fraction of acetyl, but rather to structural features, such as a more compact surface structure with high density and low cellulose accessibility. The relatively high sugar yields from both untreated and pretreated aspen suggest that aspen wood is well suited as feedstock for production of liquid biofuels and green chemicals in forest-based biorefineries.

  • 8.
    Wang, Zhao
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Wu, Guochao
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Jönsson, Leif J.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Effects of impregnation of softwood with sulfuric acid and sulfur dioxide on chemical and physical characteristics, enzymatic digestibility, and fermentability2018Ingår i: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 247, s. 200-208Artikel i tidskrift (Refereegranskat)
    Abstract [en]

    Hydrothermal pretreatment improves bioconversion of lignocellulose, but the effects of different acid catalysts are poorly understood. The effects of sulfuric acid (SA) and sulfur dioxide (SD) in continuous steam pretreatment of wood of Norway spruce were compared in the temperature range 195 degrees C-215 degrees C. The inhibitory effects of the pretreatment liquid on cellulolytic enzymes and Saccharomyces cerevisiae yeast were higher for SD-than for SApretreated material, and the inhibitory effects increased with increasing pretreatment temperature. However, the susceptibility to cellulolytic enzymes of wood pretreated with SD was 2.0-2.9 times higher than that of wood pretreated with SA at the same temperature. Data conclusively show that the superior convertibility of SDpretreated material was not due to inhibition phenomena but rather to the greater capability of the SD pretreatment to reduce the particle size through partial delignification and cellulose degradation. Particle size was shown to be correlated with enzymatic digestibility (R-2 0.97-0.98).

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