| 1. |
- Donev, Evgeniy N., et al.
(författare)
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Field testing of transgenic aspen from large greenhouse screening identifies unexpected winners
- 2023
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Ingår i: Plant Biotechnology Journal. - : Wiley. - 1467-7644 .- 1467-7652. ; 21:5, s. 1005-1021
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Tidskriftsartikel (refereegranskat)abstract
- Trees constitute promising renewable feedstocks for biorefinery using biochemical conversion, but their recalcitrance restricts their attractiveness for the industry. To obtain trees with reduced recalcitrance, large-scale genetic engineering experiments were performed in hybrid aspen blindly targeting genes expressed during wood formation and 32 lines representing seven constructs were selected for characterization in the field. Here we report phenotypes of five-year old trees considering 49 traits related to growth and wood properties. The best performing construct considering growth and glucose yield in saccharification with acid pretreatment had suppressed expression of the gene encoding an uncharacterized 2-oxoglutarate-dependent dioxygenase (2OGD). It showed minor changes in wood chemistry but increased nanoporosity and glucose conversion. Suppressed levels of SUCROSE SYNTHASE, (SuSy), CINNAMATE 4-HYDROXYLASE (C4H) and increased levels of GTPase activating protein for ADP-ribosylation factor ZAC led to significant growth reductions and anatomical abnormalities. However, C4H and SuSy constructs greatly improved glucose yields in saccharification without and with pretreatment, respectively. Traits associated with high glucose yields were different for saccharification with and without pretreatment. While carbohydrates, phenolics and tension wood contents positively impacted the yields without pretreatment and growth, lignin content and S/G ratio were negative factors, the yields with pretreatment positively correlated with S lignin and negatively with carbohydrate contents. The genotypes with high glucose yields had increased nanoporosity and mGlcA/Xyl ratio, and some had shorter polymers extractable with subcritical water compared to wild-type. The pilot-scale industrial-like pretreatment of best-performing 2OGD construct confirmed its superior sugar yields, supporting our strategy.
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| 2. |
- Alriksson, Björn, et al.
(författare)
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Ammonium hydroxide detoxification of spruce acid hydrolysates
- 2005
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Ingår i: Applied Biochemistry and Biotechnology. - 0273-2289 .- 1559-0291. ; 121, s. 911-922
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Tidskriftsartikel (refereegranskat)abstract
- When dilute-acid hydrolysates from spruce are fermented to produce ethanol, detoxification is required to make the hydrolysates fermentable at reasonable rates. Treatment with alkali, usually by overliming, is one of the most efficient approaches. Several nutrients, such as ammonium and phosphate, are added to the hydrolysates prior to fermentation. We investigated the use of NH4OH for simultaneous detoxification and addition of nitrogen source. Treatment with N-H4OH compared favorably with Ca(OH)(2), Mg(OH)(2), Ba(OH)(2), and NaOH to improve fermentability using Saccharomyces cerevisiae. Analysis of monosaccharides, furan aldehydes, phenols, and aliphatic acids was performed after the different treatments. The NH4OH treatments, performed at pH 10.0, resulted in a substantial decrease in the concentrations of furfural and hydroxymethylfurfural. Under the conditions studied, NH4OH treatments gave better results than Ca(OH)(2) treatments. The addition of an extra nitrogen source in the form of NH4Cl at pH 5.5 did not result in any improvement in fermentability that was comparable to NH4OH treatments at alkaline conditions. The addition of CaCl2 or NH4Cl at pH 5.5 after treatment with NH4OH or Ca(OH)(2) resulted in poorer fermentability, and the negative effects were attributed to salt stress. The results strongly suggest that the highly positive effects of NH4OH treatments are owing to chemical conversions rather than stimulation of the yeast cells by ammonium ions during the fermentation.
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| 3. |
- Alriksson, Björn, et al.
(författare)
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Cellulase production from spent lignocellulose hydrolysates by recombinant aspergillus niger
- 2009
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Ingår i: Applied and Environmental Microbiology. - : American Society for Microbiology. - 0099-2240 .- 1098-5336. ; 75:8, s. 2366-2374
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Tidskriftsartikel (refereegranskat)abstract
- A recombinant Aspergillus niger strain expressing the Hypocrea jecorina endoglucanase Cel7B was grown on spent hydrolysates (stillage) from sugarcane bagasse and spruce wood. The spent hydrolysates served as excellent growth media for the Cel7B-producing strain, A. niger D15[egI], which displayed higher endoglucanase activities in the spent hydrolysates than in standard medium with a comparable monosaccharide content (e.g., 2,100 nkat/ml in spent bagasse hydrolysate compared to 480 nkat/ml in standard glucose-based medium). In addition, A. niger D15[egI] was also able to consume or convert other lignocellulose-derived compounds, such as acetic acid, furan aldehydes, and phenolic compounds, which are recognized as inhibitors of yeast during ethanolic fermentation. The results indicate that enzymes can be produced from the stillage stream as a high-value coproduct in secondgeneration bioethanol plants in a way that also facilitates recirculation of process water.
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| 4. |
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| 5. |
- Alriksson, Björn
(författare)
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Ethanol from lignocellulose : Management of by-products of hydrolysis
- 2009
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)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.
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| 6. |
- Alriksson, Björn, 1979-
(författare)
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Ethanol from lignocellulose : Alkali detoxification of dilute-acid spruce hydrolysates
- 2006
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Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- 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.
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| 7. |
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| 8. |
- Alriksson, Björn, et al.
(författare)
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Fish feed from wood
- 2014
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Ingår i: Cellulose Chemistry and Technology. - 0576-9787. ; 48, s. 843-848
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Tidskriftsartikel (refereegranskat)abstract
- Increased demand of fish in combination with overexploitation of the fish stocks of the oceans has led to an increased production of fish through aquaculture. Today, fishmeal is the main protein source in fish feed for most aquaculture species. However, fishmeal is soon expected to fall short of demand as its production is associated with environmental problems. This shortage must therefore be met by sustainable alternative protein sources. Protein-rich microorganisms (i.e. Single cell protein) is an interesting option as a fishmeal substitute in fish feed which, in addition, can be produced as an important co-product in wood-based biorefineries. In the current study, four different microorganisms were cultivated on five different residual streams from Swedish wood-based biorefineries. Screening experiments were carried out in shake flasks, optimization experiments in benchtop bioreactors, and scale-up experiments were performed in a 50-litre pilot bioreactor. In addition, a demo-scale experiment was carried out in the Swedish Biorefinery Demo Plant. Microbial biomass from the scale-up experiments was collected and used for production of different fish feed formulations which, in turn, were used in feeding trials of the freshwater fish Tilapia. Fishes fed with feed, in which part of the fishmeal had been substituted with Single cell protein, showed similar or better growth than fishes fed with a fishmeal-based control feed.
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| 9. |
- Alriksson, Björn, et al.
(författare)
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Improving the fermentability of enzymatic hydrolysates of lignocellulose through chemical in-situ detoxification with reducing agents
- 2011
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Ingår i: Bioresource Technology. - : Elsevier Ltd. - 0960-8524 .- 1873-2976. ; 102:2, s. 1254-1263
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Tidskriftsartikel (refereegranskat)abstract
- Inhibitory lignocellulose hydrolysates were treated with the reducing agents dithionite and sulfite to achieve improved fermentability. Addition of these reducing agents (in the concentration range 5.0-17.5mM) to enzymatic hydrolysates of spruce wood or sugarcane bagasse improved processes based on both SHF (simultaneous hydrolysis and fermentation) and SSF (simultaneous saccharification and fermentation). The approach was exemplified in ethanolic fermentations with Saccharomyces cerevisiae and by using hydrolysates with sugar concentrations >100g/L (for SHF) and with 10% dry-matter content (for SSF). In the SHF experiments, treatments with dithionite raised the ethanol productivities of the spruce hydrolysate from 0.2 to 2.5g×L(-1)×h(-1) and of the bagasse hydrolysate from 0.9 to 3.9g×L(-1)×h(-1), values even higher than those of fermentations with reference sugar solutions without inhibitors. Benefits of the approach include that the addition of the reducing agent can be made in-situ directly in the fermentation vessel, that the treatment can be performed at a temperature and pH suitable for fermentation, and that the treatment results in dramatically improved fermentability without degradation of fermentable sugars. The many benefits and the simplicity of the approach offer a new way to achieve more efficient manufacture of fermentation products from lignocellulose hydrolysates.
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| 10. |
- Alriksson, Björn, et al.
(författare)
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Optimal conditions for alkaline detoxification of dilute-acid lignocellulose hydrolysates.
- 2006
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Ingår i: Applied Biochemistry and Biotechnology. - 0273-2289 .- 1559-0291. ; 129-132, s. 599-611
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Tidskriftsartikel (refereegranskat)abstract
- Alkaline detoxification strongly improves the fermentability of dilute-acid hydrolysates in the production of bioethanol from lignocellulose with Saccharomyces cerevisiae. New experiments were performed with NH4OH and NaOH to define optimal conditions for detoxification and make a comparison with Ca(OH)2 treatment feasible. As too harsh conditions lead to sugar degradation, the detoxification treatments were evaluated through the balanced ethanol yield, which takes both the ethanol production and the loss of fermentable sugars into account. The optimization treatments were performed as factorial experiments with 3-h duration and varying pH and temperature. Optimal conditions were found roughly in an area around pH 9.0/60 degrees C for NH4OH treatment and in a narrow area stretching from pH 9.0/80 degrees C to pH 12.0/30 degrees C for NaOH treatment. By optimizing treatment with NH4OH, NaOH, and Ca(OH)2, it was possible to find conditions that resulted in a fermentability that was equal or better than that of a reference fermentation of a synthetic sugar solution without inhibitors, regardless of the type of alkali used. The considerable difference in the amount of precipitate generated after treatment with different types of alkali appears critical for industrial implementation.
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| 11. |
- Alriksson, Björn, et al.
(författare)
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Overexpression of Saccharomyces cerevisiae transcription factor and multidrug resistance genes conveys enhanced resistance to lignocellulose-derived fermentation inhibitors
- 2010
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Ingår i: Process Biochemistry. - : Elsevier. - 1359-5113 .- 1873-3298. ; 45:2, s. 264-271
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Tidskriftsartikel (refereegranskat)abstract
- The presence of fermentation inhibitors in lignocellulose hydrolysates is an obstacle for achieving efficient fermentation of lignocellulose hydrolysates to ethanol and other commodities. In this investigation, the possibility of generating more inhibitor-resistant Saccharomyces cerevisiae by genetic engineering was explored. Based on previous results from studies of deletion mutants, three S. cerevisiae genes (ATR1, FLR1, YAP1) involved in multidrug resistance and stress response of yeast were selected for overexpression in three S. cerevisiae strains. The resistance of the transformed strains to lignocellulose-derived fermentation inhibitors and a dilute-acid spruce hydrolysate was evaluated in fermentation experiments. Overexpression of FLR1 resulted in enhanced resistance to the phenolic inhibitor coniferyl aldehyde and the furan aldehyde HMF (5-hydroxymethyl-2-furaldehyde). Overexpression of ATR1 conferred increased resistance to coniferyl aldehyde. Strains overexpressing YAP1, which encodes a transcription factor, displayed increased resistance to coniferyl aldehyde, HMF, and the spruce hydrolysate. An ethanol productivity of 0.17 g ethanol × l−1 × h−1 was achieved for a YAP1-overexpressing transformant cultivated in spruce hydrolysate, whereas a control transformant, which did not overexpress YAP1, only reached a productivity of 0.05 g ethanol × l−1 × h−1.
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| 12. |
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| 13. |
- Bergman, Kristina, et al.
(författare)
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Environmental and biodiversity performance of a novel single cell protein for rainbow trout feed
- 2024
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Ingår i: Science of the Total Environment. - : Elsevier B.V.. - 0048-9697 .- 1879-1026. ; 907
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Tidskriftsartikel (refereegranskat)abstract
- Seafood has an important role to play to achieve a sustainable food system that provides healthy food to a growing world population. Future seafood production will be increasingly reliant on aquaculture where feed innovation is essential to reduce environmental impacts and minimize feed and food competition. This study aimed to investigate whether a novel single cell protein feed ingredient based on Paecilomyces variotii grown on a side stream from the forest industry could improve environmental sustainability of farmed rainbow trout (Oncorhynchus mykiss) by replacing the soy protein concentrate used today. A Life Cycle Assessment including commonly addressed impacts but also the rarely assessed biodiversity impacts was performed. Furthermore, feeding trials were included for potential effects on fish growth, i.e., an assessment of the environmental impacts for the functional unit ‘kg feed required to produce 1 kg live-weight rainbow trout’. Results showed that the best experimental diet containing P. variotii performed 16–73 % better than the control diet containing soy protein concentrate in all impact categories except for energy demand (21 % higher impact). The largest environmental benefits from replacing soy protein with P. variotii in rainbow trout diets was a 73 % reduction of impact on biodiversity and halved greenhouse gas emissions. The findings have high relevance for the aquaculture industry as the production scale and feed composition was comparable to commercial operations and because the effect on fish growth from inclusion of the novel ingredient in a complete diet was evaluated. The results on biodiversity loss from land use change and exploitation through fishing suggest that fishery can dominate impacts and exclusion thereof can greatly underestimate biodiversity impact. Finally, a novel feed ingredient grown on side streams from the forest industry has potential to add to food security through decreasing the dependence on increasingly scarce agricultural land resources.
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| 14. |
- Cavka, Adnan, et al.
(författare)
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Biorefining of wood : combined production of ethanol and xylanase from waste fiber sludge
- 2011
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Ingår i: Journal of Industrial Microbiology & Biotechnology. - : Springer. - 1367-5435 .- 1476-5535. ; 38:8, s. 891-899
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Tidskriftsartikel (refereegranskat)abstract
- The possibility to utilize fiber sludge, waste fibers from pulp mills and lignocellulose-based biorefineries, for combined production of liquid biofuel and biocatalysts was investigated. Without pretreatment, fiber sludge was hydrolyzed enzymatically to monosaccharides, mainly glucose and xylose. In the first of two sequential fermentation steps, the fiber sludge hydrolysate was fermented to cellulosic ethanol with the yeast Saccharomyces cerevisiae. Although the final ethanol yields were similar, the ethanol productivity after 9.5 h was 3.3 g/l/h for the fiber sludge hydrolysate compared with only 2.2 g/l/h for a reference fermentation with similar sugar content. In the second fermentation step, the spent fiber sludge hydrolysate (the stillage obtained after distillation) was used as growth medium for recombinant Aspergillus niger expressing the xylanase-encoding Trichoderma reesei (Hypocrea jecorina) xyn2 gene. The xylanase activity obtained with the spent fiber sludge hydrolysate (8,500 nkat/ml) was higher than that obtained in a standard medium with similar monosaccharide content (1,400 nkat/ml). Analyses based on deglycosylation with N-glycosidase F suggest that the main part of the recombinant xylanase was unglycosylated and had molecular mass of 20.7 kDa, while a minor part had N-linked glycosylation and molecular mass of 23.6 kDa. Chemical analyses of the growth medium showed that important carbon sources in the spent fiber sludge hydrolysate included xylose, small aliphatic acids, and oligosaccharides. The results show the potential of converting waste fiber sludge to liquid biofuel and enzymes as coproducts in lignocellulose-based biorefineries.
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| 15. |
- Cavka, Adnan, et al.
(författare)
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Effect of sulfur oxyanions on lignocellulose-derived fermentation inhibitors
- 2011
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Ingår i: Biotechnology and Bioengineering. - : Wiley. - 0006-3592 .- 1097-0290. ; 108:11, s. 2592-2599
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Tidskriftsartikel (refereegranskat)abstract
- Recent results show that treatments with reducing agents, including the sulfur oxyanions dithionite and hydrogen sulfite, efficiently improve the fermentability of inhibitory lignocellulose hydrolysates, and that the treatments are effective when the reducing agents are added in situ into the fermentation vessel at low temperature. In the present investigation, dithionite was added to medium with model inhibitors (coniferyl aldehyde, furfural, 5-hydroxymethylfurfural, or acetic acid) and the effects on the fermentability with yeast were studied. Addition of 10 mM dithionite to medium containing 2.5 mM coniferyl aldehyde resulted in a nine-fold increase in the glucose consumption rate and a three-fold increase in the ethanol yield. To investigate the mechanism behind the positive effects of adding sulfur oxyanions, mixtures containing 2.5 mM of a model inhibitor (an aromatic compound, a furan aldehyde, or an aliphatic acid) and 15 mM dithionite or hydrogen sulfite were analyzed using mass spectrometry (MS). The results of the analyses, which were performed by using UHPLC-ESI-TOF-MS and UHPLC-LTQ/Orbitrap-MS/MS, indicate that the positive effects of sulfur oxyanions are primarily due to their capability to react with and sulfonate inhibitory aromatic compounds and furan aldehydes at low temperature and slightly acidic pH (such as 25°C and pH 5.5).
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| 16. |
- Cavka, Adnan, et al.
(författare)
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Ozone detoxification of steam-pretreated Norway spruce
- 2015
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Ingår i: Biotechnology for Biofuels. - : BioMed Central. - 1754-6834. ; 8
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Tidskriftsartikel (refereegranskat)abstract
- 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 > 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.
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| 17. |
- Cavka, Adnan, et al.
(författare)
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Production of cellulosic ethanol and enzyme from waste fiber sludge using SSF, recycling of hydrolytic enzymes and yeast, and recombinant cellulase-producing Aspergillus niger
- 2014
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Ingår i: Journal of Industrial Microbiology & Biotechnology. - : Springer Berlin/Heidelberg. - 1367-5435 .- 1476-5535. ; 41:8, s. 1191-1200
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Tidskriftsartikel (refereegranskat)abstract
- Bioethanol and enzymes were produced from fiber sludges through sequential microbial cultivations. After a first simultaneous saccharification and fermentation (SSF) with yeast, the bioethanol concentrations of sulfate and sulfite fiber sludges were 45.6 and 64.7 g/L, respectively. The second SSF, which included fresh fiber sludges and recycled yeast and enzymes from the first SSF, resulted in ethanol concentrations of 38.3 g/L for sulfate fiber sludge and 24.4 g/L for sulfite fiber sludge. Aspergillus niger carrying the endoglucanase-encoding Cel7B gene of Trichoderma reesei was grown in the spent fiber sludge hydrolysates. The cellulase activities obtained with spent hydrolysates of sulfate and sulfite fiber sludges were 2,700 and 2,900 nkat/mL, respectively. The high cellulase activities produced by using stillage and the significant ethanol concentrations produced in the second SSF suggest that onsite enzyme production and recycling of enzyme are realistic concepts that warrant further attention.
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| 18. |
- Chen, Genqiang, et al.
(författare)
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Bioconversion of waste fiber sludge to bacterial nanocellulose and use for reinforcement of CTMP paper sheets
- 2017
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Ingår i: Polymers. - : MDPI AG. - 2073-4360. ; 9:9
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Tidskriftsartikel (refereegranskat)abstract
- 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 × 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 × 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.
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| 19. |
- Chen, Genqiang, et al.
(författare)
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Scale-up of production of bacterial nanocellulose using submerged cultivation
- 2018
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Ingår i: Journal of chemical technology and biotechnology (1986). - : Wiley. - 0268-2575 .- 1097-4660. ; 93:12, s. 3418-3427
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: More extensive utilization of bacterial nanocellulose (BNC) is severely restricted by the low efficiency and small scale of the traditional static cultivation. Submerged fermentation in stirred-tank reactors (STRs) is potentially favourable for large-scale production of BNC, but scale-up of cultivation remains challenging. Even though the STR is most commonly used for submerged cultivation in the fermentation industry, there are few previous attempts to scale-up production of BNC to pilot scale using an STR. Furthermore, the question of how scale-up of submerged cultivation affects the properties of the BNC has received very little attention. RESULTS: Four strains were compared in 250-mL shake flasks. Strain DHU-ATCC-1 displayed the highest volumetric productivity, 0.56 g L−1 d−1, and was then cultivated in a 400-mL STR, showing a similar productivity of 0.55 g L−1 d−1. Scale-up using a 75-L STR pilot bioreactor resulted in enhancement of the BNC production rate from 0.056 g d−1 in the shake flasks to 17.3 g d−1 in the 75-L STR, although the productivity decreased to 0.43 g L−1 d−1. During scale-up from shake flasks to 400-mL STR and further on to 75-L STR, the BNC fibers formed more bundles, whereas the fiber diameter decreased from 25.6 to 21.7 nm. The BNC from the 75-L STR exhibited a higher degree of polymerization, specifically 3230, higher degree of crystallinity, specifically 83%, larger crystallites, and improved strength including higher tensile energy absorption index and superior stretch at break. CONCLUSION: It is possible to enhance BNC production, and maintain or improve its properties when scaling up submerged cultivation in STRs.
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| 20. |
- Ginesy, Mireille
(författare)
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Production of L-arginine by genetically modified Escherichia coli
- 2015
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Licentiatavhandling (övrigt vetenskapligt/konstnärligt)abstract
- In the recent years, the demand for environmental friendly produced L-arginine has risen with the increasing number of applications for this amino acid in pharmaceuticals, nutraceuticals, cosmetics, animal feed and fertilizers. Members of the Corynebacteriaceae family are usually used for microbial L-arginine production. However, Escherichia coli present the advantage of being able to utilize a wider range of substrates, including pentose sugars found in lignocellulosic feedstocks. The present thesis illustrates the first steps in the development of a sustainable process to produce L-arginine using E. coli. It starts with the constructions of a L-arginine overproducing strain, followed by optimization of the nitrogen supply for the fermentations.The first part of this thesis aimed at engineering an E. coli train able to produce high level of L-arginine. Mutations on key genes of the L-arginine biosynthesis pathway were step-wisely done. The mutants obtained at each step were tested in bioreactor fermentations to assess the effect of each genetic modification. The final strain was able to produce almost 12 g/l during fermentation, at a productivity of 0.24 g/l/h. In comparison the starting strain, E. coli K12 C600, was not able to excrete any L-arginine. To minimize nitrogen wastes and optimize the L-arginine production the impact of different nitrogen sources and concentration were investigated. It was shown that while ammonium phosphate dibasic was the most potent nitrogen source during cultivation on complex medium, all the sources were equivalent with minimal media; this probably reflected the phosphate deficiency of the complex medium used. In fermentation on minimal medium, a carbon to nitrogen ratio of 5 was demonstrated to be the most suitable, yielding up to 4.5 g/l L-arginine. At this ratio, both glucose and the nitrogen source were completely utilized during fermentation.
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| 21. |
- Ilanidis, Dimitrios, 1986-
(författare)
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Biochemical conversion of biomass : hydrothermal pretreatment, by-product formation, conditioning, enzymatic saccharification, and fermentability
- 2021
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Lignocellulosic residues have great potential as feedstocks for production of bio-based chemicals and fuels. One of the main routes is biochemical conversion, which typically includes pretreatment, enzymatic saccharification, microbial fermentation of sugars, and valorization of hydrolysis lignin. Utilization of a broad variety of lignocellulosic feedstocks and development of more efficient conversion processes are advantageous for making bio-based commodities affordable. Biochemical conversion of lignocellulosic biomass was investigated with the overall aim to understand how variations in pretreatment conditions affected yields, formation of bioconversion inhibitors, enzymatic digestibility of pretreated materials, and fermentability. Experiments included estimation of pseudo-lignin content and quantitation of recently discovered microbial inhibitors, such as formaldehyde and p-benzoquinone. Conditioning of pretreated material to improve the efficiency of reactions with biocatalysts was further investigated.Hydrothermal pretreatment of sugarcane bagasse was investigated by using both autocatalyzed and sulfuric-acid-catalyzed pretreatment and by varying temperature and time in such a way that the severity factor was maintained at one of three predetermined values. For autocatalyzed pretreatments, the enzymatic digestibility of the pretreated solids was directly proportional to the severity. Pretreatment conditions that were just harsh enough to almost quantitatively solubilize hemicelluloses gave the best results. Potential effects of the redox environment during hydrothermal pretreatment were investigated by addition of oxygen gas or nitrogen gas in experiments with sugarcane bagasse and Norway spruce. The investigation demonstrated that gas addition, and especially addition of oxygen gas, can be used to modulate the severity of acidic hydrothermal pretreatment. Hydrothermal pretreatment of wheat straw was investigated to evaluate the impact of pretreatment conditions on newly discovered inhibitors, enzymatic digestibility, and fermentation. An increase of the temperature up to 190 °C in autocatalyzed pretreatment led to high combined glucose and xylose yields; up to ~480 kg/ton (dry weight) raw wheat straw. A correlation between enzymatic digestibility and removal of hemicelluloses was observed. A techno-economical evaluation of several conditioning methods for slurries of steam-pretreated spruce indicated that treatment with sodium sulfite was the most favorable option. Treatments with sulfite and dithionite successfully decreased the concentration of formaldehyde. Results also indicate that increased temperature in conditioning of hydrolysate could to some extent compensate for using lower dosages of sodium dithionite.
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| 22. |
- Ilanidis, Dimitrios, et al.
(författare)
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Comparison of Efficiency and Cost of Methods for Conditioning of Slurries of Steam-Pretreated Softwood
- 2021
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Ingår i: Frontiers in Energy Research. - : Frontiers Media S.A.. - 2296-598X. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- 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.
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| 23. |
- Ilanidis, Dimitrios, et al.
(författare)
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Factors affecting detoxification of softwood enzymatic hydrolysates using sodium dithionite
- 2021
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Ingår i: Processes. - : MDPI AG. - 2227-9717. ; 9:5
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Tidskriftsartikel (refereegranskat)abstract
- 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. © 2021 by the authors.
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| 24. |
- Jönsson, Leif J., et al.
(författare)
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Bioconversion of lignocellulose : Inhibitors and detoxification
- 2013
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Ingår i: Biotechnology for Biofuels. - : Springer Science and Business Media LLC. - 1754-6834. ; 6:1
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Tidskriftsartikel (refereegranskat)abstract
- 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.
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| 25. |
- Jönsson, Leif, et al.
(författare)
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Methods for improvement of enzymatic hydrolysis of lignocellullosic material
- 2013
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Patent (populärvet., debatt m.m.)abstract
- 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)
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