| 1. |
- Nickel, David, 1990
(författare)
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Process development for platform chemical production from agricultural and forestry residues
- 2021
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- As part of a bio-based economy, biorefineries are envisaged to sustainably produce platform chemicals via biochemical conversion of agricultural and forestry residues. However, supply risks, the recalcitrance of lignocellulosic biomass, and inhibitor formation during pretreatment impair the economic feasibility of such biorefineries. In this thesis, process design and assessment were developed with the aim of addressing these hurdles and improving the cost-effectiveness of lignocellulose-derived platform chemicals. To expand the feedstock base and reduce operational costs, logging residues served as underutilised and inexpensive raw material. The major impediment in converting logging residues was their high recalcitrance and low cellulose content, which resulted in low attainable ethanol titres during simultaneous saccharification and co-fermentation (SSCF). Pretreatment optimisation reduced inhibitor formation and recalcitrance, and led to enzymatic hydrolysis yields at par with those obtained for stem wood, despite the less favourable chemical composition. Upgrading logging residues with carbohydrate-rich oat hulls increased ethanol titres to >50 g/L using batch SSCF at 20% WIS loadings, demonstrating the potential to further decrease downstream processing costs. To alleviate the toxicity of inhibitors generated during pretreatment, preadaptation was applied to Saccharomyces cerevisiae . Exposure to the inhibitors in the pretreated liquid fraction improved ethanol production during subsequent fermentation. Transferring the concept of preadaptation to lactic acid production by Bacillus coagulans cut the process times by half and more than doubled the average specific lactic acid productivity, showcasing how preadaptation could decrease operational costs. To assess the performance and robustness of process designs against process input variations, a multi-scale variability analysis framework was developed. The framework included models for bioprocess, flowsheet, techno-economic, and life cycle assessment. In a case study, multi-feed processes, in which solids and cells are fed to the process using model-based predictions, were more robust against variable cellulolytic activities than batch SSCFs in a wheat straw-based ethanol biorefinery. The developed framework can be used to identify robust biorefinery process designs, which simultaneously meet technological, economic, and environmental goals.
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| 2. |
- Borgström, Celina, et al.
(författare)
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Using phosphoglucose isomerase-deficient (pgi1Δ) Saccharomyces cerevisiae to map the impact of sugar phosphate levels on d-glucose and d-xylose sensing
- 2022
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Ingår i: Microbial Cell Factories. - : Springer Science and Business Media LLC. - 1475-2859. ; 21:1
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Tidskriftsartikel (refereegranskat)abstract
- Background: Despite decades of engineering efforts, recombinant Saccharomyces cerevisiae are still less efficient at converting d-xylose sugar to ethanol compared to the preferred sugar d-glucose. Using GFP-based biosensors reporting for the three main sugar sensing routes, we recently demonstrated that the sensing response to high concentrations of d-xylose is similar to the response seen on low concentrations of d-glucose. The formation of glycolytic intermediates was hypothesized to be a potential cause of this sensing response. In order to investigate this, glycolysis was disrupted via the deletion of the phosphoglucose isomerase gene (PGI1) while intracellular sugar phosphate levels were monitored using a targeted metabolomic approach. Furthermore, the sugar sensing of the PGI1 deletants was compared to the PGI1-wildtype strains in the presence of various types and combinations of sugars. Results: Metabolomic analysis revealed systemic changes in intracellular sugar phosphate levels after deletion of PGI1, with the expected accumulation of intermediates upstream of the Pgi1p reaction on d-glucose and downstream intermediates on d-xylose. Moreover, the analysis revealed a preferential formation of d-fructose-6-phosphate from d-xylose, as opposed to the accumulation of d-fructose-1,6-bisphosphate that is normally observed when PGI1 deletants are incubated on d-fructose. This may indicate a role of PFK27 in d-xylose sensing and utilization. Overall, the sensing response was different for the PGI1 deletants, and responses to sugars that enter the glycolysis upstream of Pgi1p (d-glucose and d-galactose) were more affected than the response to those entering downstream of the reaction (d-fructose and d-xylose). Furthermore, the simultaneous exposure to sugars that entered upstream and downstream of Pgi1p (d-glucose with d-fructose, or d-glucose with d-xylose) resulted in apparent synergetic activation and deactivation of the Snf3p/Rgt2p and cAMP/PKA pathways, respectively. Conclusions: Overall, the sensing assays indicated that the previously observed d-xylose response stems from the formation of downstream metabolic intermediates. Furthermore, our results indicate that the metabolic node around Pgi1p and the level of d-fructose-6-phosphate could represent attractive engineering targets for improved d-xylose utilization.
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| 3. |
- Brink, Daniel P., et al.
(författare)
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D-xylose sensing in saccharomyces cerevisiae : Insights from D-glucose signaling and native D-xylose utilizers
- 2021
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Ingår i: International Journal of Molecular Sciences. - : MDPI AG. - 1661-6596 .- 1422-0067. ; 22:22
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Forskningsöversikt (refereegranskat)abstract
- Extension of the substrate range is among one of the metabolic engineering goals for microorganisms used in biotechnological processes because it enables the use of a wide range of raw materials as substrates. One of the most prominent examples is the engineering of baker’s yeast Saccharomyces cerevisiae for the utilization of D-xylose, a five-carbon sugar found in high abundance in lignocellulosic biomass and a key substrate to achieve good process economy in chemical production from renewable and non-edible plant feedstocks. Despite many excellent engineering strategies that have allowed recombinant S. cerevisiae to ferment D-xylose to ethanol at high yields, the consumption rate of D-xylose is still significantly lower than that of its preferred sugar D-glucose. In mixed D-glucose/D-xylose cultivations, D-xylose is only utilized after D-glucose depletion, which leads to prolonged process times and added costs. Due to this limitation, the response on D-xylose in the native sugar signaling pathways has emerged as a promising next-level engineering target. Here we review the current status of the knowledge of the response of S. cerevisiae signaling pathways to D-xylose. To do this, we first summarize the response of the native sensing and signaling pathways in S. cerevisiae to D-glucose (the preferred sugar of the yeast). Using the Dglucose case as a point of reference, we then proceed to discuss the known signaling response to Dxylose in S. cerevisiae and current attempts of improving the response by signaling engineering using native targets and synthetic (non-native) regulatory circuits.
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| 4. |
- Persson, Viktor C., et al.
(författare)
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Impact of xylose epimerase on sugar assimilation and sensing in recombinant Saccharomyces cerevisiae carrying different xylose-utilization pathways
- 2023
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Ingår i: Biotechnology for Biofuels and Bioproducts. - 2731-3654. ; 16:1
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Tidskriftsartikel (refereegranskat)abstract
- BackgroundOver the last decades, many strategies to procure and improve xylose consumption in Saccharomyces cerevisiae have been reported. This includes the introduction of efficient xylose-assimilating enzymes, the improvement of xylose transport, or the alteration of the sugar signaling response. However, different strain backgrounds are often used, making it difficult to determine if the findings are transferrable both to other xylose-consuming strains and to other xylose-assimilation pathways. For example, the influence of anomerization rates between α- and β-xylopyranose in pathway optimization and sugar sensing is relatively unexplored.ResultsIn this study, we tested the effect of expressing a xylose epimerase in S. cerevisiae strains carrying different xylose-consuming routes. First, XIs originating from three different species in isogenic S. cerevisiae strains were tested and the XI from Lachnoclostridium phytofermentans was found to give the best performance. The benefit of increasing the anomerization rate of xylose by adding a xylose epimerase to the XI strains was confirmed, as higher biomass formation and faster xylose consumption were obtained. However, the impact of xylose epimerase was XI-dependent, indicating that anomer preference may differ from enzyme to enzyme. The addition of the xylose epimerase in xylose reductase/xylitol dehydrogenase (XR/XDH)-carrying strains gave no improvement in xylose assimilation, suggesting that the XR from Spathaspora passalidarum had no anomer preference, in contrast to other reported XRs. The reduction in accumulated xylitol that was observed when the xylose epimerase was added may be associated with the upregulation of genes encoding endogenous aldose reductases which could be affected by the anomerization rate. Finally, xylose epimerase addition did not affect the sugar signaling, whereas the type of xylose pathway (XI vs. XR/XDH) did.ConclusionsAlthough xylose anomer specificity is often overlooked, the addition of xylose epimerase should be considered as a key engineering step, especially when using the best-performing XI enzyme from L. phytofermentans. Additional research into the binding mechanism of xylose is needed to elucidate the enzyme-specific effect and decrease in xylitol accumulation. Finally, the differences in sugar signaling responses between XI and XR/XDH strains indicate that either the redox balance or the growth rate impacts the SNF1/Mig1p sensing pathway.
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| 5. |
- Amiandamhen, Stephen, 1983-, et al.
(författare)
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Bioenergy production and utilization in different sectors in Sweden: A state of the art review
- 2020
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Ingår i: BioResources. - : University of North Carolina Press. - 1930-2126. ; 15:4, s. 9834-9857
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Forskningsöversikt (refereegranskat)abstract
- In the continual desire to reduce the environmental footprints of human activities, research efforts to provide cleaner energy is increasingly becoming vital. The effect of climate change on present and future existence, sustainable processes, and utilizations of renewable resources have been active topics within international discourse. In order to reduce the greenhouse gases emissions from traditional materials and processes, there has been a shift to more environmental friendly alternatives. The conversion of biomass to bioenergy, including biofuels has been considered to contribute to the future of climate change mitigation, although there are concerns about carbon balance from forest utilization. Bioenergy accounts for more than one-third of all energy used in Sweden and biomass has provided about 60% of the fuel for district heating. Apart from heat and electricity supply, the transport sector, with about 30% of global energy use, has a significant role in a sustainable bioenergy system. This review presents the state of the art in the Swedish bioenergy sector based on literature and Swedish Energy Agency’s current statistics. The review also discusses the overall bioenergy production and utilization in different sectors in Sweden. The current potential, challenges, and environmental considerations of bioenergy production are also discussed.
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| 6. |
- Kawde, Anurag, et al.
(författare)
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Photoelectrochemical oxidation in ambient conditions using earth-abundant hematite anode : A green route for the synthesis of biobased polymer building blocks
- 2021
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Ingår i: Catalysts. - : MDPI AG. - 2073-4344. ; 11:8
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Tidskriftsartikel (refereegranskat)abstract
- This study demonstrates the use of a photoelectrochemical device comprising earth-abundant hematite photoanode for the oxidation of 5-hydroxymethylfurfural (5-HMF), a versatile bio-based platform chemical, under ambient conditions in the presence of an electron mediator. The results obtained in this study showed that the hematite photoanode, upon doping with fluorine, can oxidize water even at lower pH (4.5 and 9.0). For 5-HMF oxidation, three different pH conditions were investigated, and complete oxidation to 2,5-furandicarboxylic acid (FDCA) via 5-hydroxymethyl-2-furancarboxylic acid (HMFCA) was achieved at pH above 12. At lower pH, the oxidation followed another route via 2,5-diformylfuran (DFF), yielding 5-formyl-2-furancarboxylic acid (FFCA) as the main product. Using the oxidized intermediates as substrates showed DFF to be most efficiently oxidized to FDCA. We also show that, at pH 4.5, the addition of the laccase enzyme promoted the oxidation of 5-HMF to FFCA.
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| 7. |
- Shin, Jae Ho, 1987, et al.
(författare)
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Synthetic pathway engineering in Corynebacterium glutamicum for production of adipic acid
- 2020
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- We are interested in production of adipic acid from renewable sources using a gram-positive bacterium, Corynebacterium glutamicum ATCC 13032, as a target for metabolic engineering. We aim to construct a synthetic pathway that would allow biosynthesis of adipic acid and to implement it into the host chassis for utilizing simple sugar, glucose, as the starting raw material. The chosen pathway stems off from intermediates from the central metabolism and requires 5 synthetic biochemical steps before reaching adipic acid. The pathway is equipped with a promiscuous enzyme to allow leakage at each metabolic step and byproduct formation in order to monitor the efficiency of each step of the pathway. In order to ensure translation, each gene introduced was codon-optimized to C. glutamicum. Introducing the synthetic genes one by one into C. glutamicum as well as flask-level cultivation of engineered strains in a semi-defined medium allowed detection of each byproduct of the pathway leading up to adipic acid by GC/MS based methods. Further, the codon-optimized version of the 5 synthetic genes were constructed in 2 separate operons each with an inducible promoter. Finally, we analyzed the translational efficiency of the genes and modified the constructs for a better operon expression. We implement additional sample preparation methods for isolating and concentrating adipic acid content for better analysis. The results shown here will be used to further develop and complete biosynthesis of adipic acid from a C. glutamicum chassis.
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| 8. |
- Abitbol, Tiffany, et al.
(författare)
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Isolation of Mixed Compositions of Cellulose Nanocrystals, Microcrystalline Cellulose, and Lignin Nanoparticles from Wood Pulps
- 2023
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Ingår i: ACS Omega. - : American Chemical Society (ACS). - 2470-1343. ; 8:24, s. 21474-21484
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Tidskriftsartikel (refereegranskat)abstract
- From a circular economyperspective, one-pot strategies for theisolation of cellulose nanomaterials at a high yield and with multifunctionalproperties are attractive. Here, the effects of lignin content (bleachedvs unbleached softwood kraft pulp) and sulfuric acid concentrationon the properties of crystalline lignocellulose isolates and theirfilms are explored. Hydrolysis at 58 wt % sulfuric acid resulted inboth cellulose nanocrystals (CNCs) and microcrystalline celluloseat a relatively high yield (>55%), whereas hydrolysis at 64 wt% gaveCNCs at a lower yield (<20%). CNCs from 58 wt % hydrolysis weremore polydisperse and had a higher average aspect ratio (1.5-2x),a lower surface charge (2x), and a higher shear viscosity (100-1000x).Hydrolysis of unbleached pulp additionally yielded spherical nanoparticles(NPs) that were <50 nm in diameter and identified as lignin bynanoscale Fourier transform infrared spectroscopy and IR imaging.Chiral nematic self-organization was observed in films from CNCs isolatedat 64 wt % but not from the more heterogeneous CNC qualities producedat 58 wt %. All films degraded to some extent under simulated sunlighttrials, but these effects were less pronounced in lignin-NP-containingfilms, suggesting a protective feature, but the hemicellulose contentand CNC crystallinity may be implicated as well. Finally, heterogeneousCNC compositions obtained at a high yield and with improved resourceefficiency are suggested for specific nanocellulose uses, for instance,as thickeners or reinforcing fillers, representing a step toward thedevelopment of application-tailored CNC grades.
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| 9. |
- Bohman, Björn
(författare)
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Seed dispersal: Hungry hornets are unexpected and effective vectors
- 2022
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Ingår i: Current Biology. - : Elsevier BV. - 0960-9822 .- 1879-0445. ; 32, s. R836-R838
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Tidskriftsartikel (refereegranskat)abstract
- A new study finds that, in the forests of tropical China, hungry hornets are lured to the fruits of Aquilaria sinensis by highly volatile compounds structurally similar to volatiles from herbivore-damaged leaves. The hornets disperse the short-lived seeds rapidly to optimal new habitats.
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| 10. |
- Choi, BoHyun, 1986, et al.
(författare)
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Engineering of Saccharomyces cerevisiae for enhanced metabolic robustness and L-lactic acid production from lignocellulosic biomass
- 2024
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Ingår i: Metabolic Engineering. - 1096-7176 .- 1096-7184. ; 84, s. 23-33
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Tidskriftsartikel (refereegranskat)abstract
- Metabolic engineering for high productivity and increased robustness is needed to enable sustainable biomanufacturing of lactic acid from lignocellulosic biomass. Lactic acid is an important commodity chemical used for instance as a monomer for production of polylactic acid, a biodegradable polymer. Here, rational and model-based optimization was used to engineer a diploid, xylose fermenting Saccharomyces cerevisiae strain to produce L-lactic acid. The metabolic flux was steered towards lactic acid through the introduction of multiple lactate dehydrogenase encoding genes while deleting ERF2, GPD1, and CYB2. A production of 93 g/L of lactic acid with a yield of 0.84 g/g was achieved using xylose as the carbon source. To increase xylose utilization and reduce acetic acid synthesis, PHO13 and ALD6 were also deleted from the strain. Finally, CDC19 encoding a pyruvate kinase was overexpressed, resulting in a yield of 0.75 g lactic acid/g sugars consumed, when the substrate used was a synthetic lignocellulosic hydrolysate medium, containing hexoses, pentoses and inhibitors such as acetate and furfural. Notably, modeling also provided leads for understanding the influence of oxygen in lactic acid production. High lactic acid production from xylose, at oxygen-limitation could be explained by a reduced flux through the oxidative phosphorylation pathway. On the contrast, higher oxygen levels were beneficial for lactic acid production with the synthetic hydrolysate medium, likely as higher ATP concentrations are needed for tolerating the inhibitors therein. The work highlights the potential of S. cerevisiae for industrial production of lactic acid from lignocellulosic biomass.
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