| 1. |
- Munthe, Christian, 1962
(author)
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Precaution and Ethics: Handling risks, uncertainties and knowledge gaps in the regulation of new biotechnologies
- 2017
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Book (other academic/artistic)abstract
- This volume outlines and analyses ethical issues actualized by applying a precautionary approach to the regulation of new biotechnologies. It presents a novel way of categorizing and comparing biotechnologies from a precautionary standpoint. Based on this, it addresses underlying philosophical problems regarding the ethical assessment of decision-making under uncertainty and ignorance, and discusses how risks and possible benefits of such technologies should be balanced from an ethical standpoint. It argues on conceptual and ethical grounds for a technology neutral regulation as well as for a regulation that not only checks new technologies but also requires old, inferior ones to be phased out. It demonstrates how difficult ethical issues regarding the extent and ambition of precautionary policies need to be handled by such a regulation, and presents an overarching framework for doing so.
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| 2. |
- Eivazihollagh, Alireza, et al.
(author)
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On chelating surfactants : Molecular perspectives and application prospects
- 2019
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In: Journal of Molecular Liquids. - : Elsevier BV. - 0167-7322 .- 1873-3166. ; 278, s. 688-705
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Journal article (peer-reviewed)abstract
- Chelating agents, molecules that very strongly coordinates certain metal ions, are used industrially as well as in consumer products to minimize disturbances and increase performance of reactions and applications. The widely used sequestering agents, nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DTPA) belong to this branch of readily water-soluble compounds. When these chemical structures also have hydrophobic parts, they are prone to adsorb at air-water interfaces and to self-assemble. Such bifunctional molecules can be called chelating surfactants and will have more extended utilization prospects than common chelating agents or ordinary ionic surfactants. The present review attempts to highlight the fundamental behavior of chelating surfactants in solution and at interfaces, and their very specific interactions with metal ions. Methods to recover chelating surfactants from metal chelates are also described. Moreover, utilization of chelating surfactants in applications for metal removal in environmental engineering and mineral processing, as well as for metal control in the fields of biology, chemistry and physics, is exemplified and discussed.
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| 3. |
- Munthe, Christian, 1962
(author)
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The Price of Precaution and the Ethics of Risk
- 2011
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Book (other academic/artistic)abstract
- Since a couple of decades, the notion of a precautionary principle plays a central and increasingly influential role in international as well as national policy and regulation regarding the environment and the use of technology. Urging society to take action in the face of potential risks of human activities in these areas, the recent focus on climate change has further sharpened the importance of this idea. However, the idea of a precautionary principle has also been problematised and criticised by scientists, scholars and policy activists, and been accused of almost every intellectual sin imaginable: unclarity, impracticality, arbitrariness and moral as well as political unsoundness. In that light, the very idea of precaution as an ideal for policy making rather comes out as a dead end. On the basis of these contrasting starting points, Christian Munthe undertakes an innovative, in-depth philosophical analysis of what the idea of a precautionary principle is and should be about. A novel theory of the ethics of imposing risks is developed and used as a foundation for defending the idea of precaution in environmental and technological policy making against its critics, while at the same time avoiding a number of identified flaws. The theory is shown to have far-reaching consequences for areas such as bio-, information- and nuclear technology, and global environmental policy in areas such as climate change. The author argues that, while the price we pay for precaution must not be too high, we have to be prepared to pay it in order to act ethically defensible. A number of practical suggestions for precautionary regulation and policy making are made on the basis of this, and some challenges to basic ethical theory as well as consumerist societies, the global political order and liberal democracy are identified
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| 4. |
- Mayers, Joshua, 1988, et al.
(author)
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Integrating Microalgal Production with Industrial Outputs - Reducing Process Inputs and Quantifying the Benefits
- 2016
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In: Industrial Biotechnology. - : Mary Ann Liebert Inc. - 1550-9087 .- 1931-8421. ; 12:4, s. 219-234
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Journal article (peer-reviewed)abstract
- The cultivation and processing of microalgal biomass is resource- and energy-intensive, negatively affecting the sustainability and profitability of producing bulk commodities, limiting this platform to the manufacture of relatively small quantities of high-value compounds. A biorefinery approach where all fractions of the biomass are valorized might improve the case for producing lower-value products. However, these systems are still likely to operate very close to thresholds of profitability and energy balance, with wide-ranging environmental and societal impacts. It thus remains critically important to reduce the use of costly and impactful inputs and energy-intensive processes involved in these scenarios. Integration with industrial infrastructure can provide a number of residual streams that can be readily used during microalgal cultivation and downstream processing. This review critically considers some of the main inputs required for microalgal biorefineries - such as nutrients, water, carbon dioxide, and heat - and appraises the benefits and possibilities for industrial integration on a more quantitative basis. Recent literature and demonstration studies will also be considered to best illustrate these benefits to both producers and industrial operators. Additionally, this review will highlight some inconsistencies in the data used in assessments of microalgal production scenarios, allowing more accurate evaluation of potential future biorefineries.
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| 5. |
- Sepehri, Sobhan, 1986, et al.
(author)
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Volume-amplified magnetic bioassay integrated with microfluidic sample handling and high-Tc SQUID magnetic readout
- 2018
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In: APL Bioengineering. - : AIP Publishing. - 2473-2877. ; 2:1
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Journal article (peer-reviewed)abstract
- A bioassay based on a high-Tc superconducting quantum interference device (SQUID) reading out functionalized magnetic nanoparticles (fMNPs) in a prototype microfluidic platform is presented. The target molecule recognition is based on volume amplification using padlock-probe-ligation followed by rolling circle amplification (RCA). The MNPs are functionalized with single-stranded oligonucleotides, which give a specific binding of the MNPs to the large RCA coil product, resulting in a large change in the amplitude of the imaginary part of the ac magnetic susceptibility. The RCA products from amplification of synthetic Vibrio cholera target DNA were investigated using our SQUID ac susceptibility system in microfluidic channel with an equivalent sample volume of 3 μl. From extrapolation of the linear dependence of the SQUID signal versus concentration of the RCA coils, it is found that the projected limit of detection for our system is about 1.0 e5 RCA coils (0.2e−18 mol), which is equivalent to 66 fM in the 3 μl sample volume. This ultra-high magnetic sensitivity and integration with microfluidic sample handling are critical steps towards magnetic bioassays for rapid detection of DNA and RNA targets at the point of care.
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| 6. |
- Hong, Kuk-ki, 1976, et al.
(author)
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Metabolic Engineering of Saccharomyces cerevisiae: A Key Cell Factory Platform for Future Biorefineries
- 2012
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In: Cellular and Molecular Life Sciences. - : Springer Science and Business Media LLC. - 1420-9071 .- 1420-682X. ; 69:16, s. 2671-2690
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Research review (peer-reviewed)abstract
- Metabolic engineering is the enabling science of development of efficient cell factories for the production of fuels, chemicals, pharmaceuticals, and food ingredients through microbial fermentations. The yeast Saccharomyces cerevisiae is a key cell factory already used for the production of a wide range of industrial products, and here we review ongoing work, particularly in industry, on using this organism for the production of butanol, which can be used as biofuel, and isoprenoids, which can find a wide range of applications including as pharmaceuticals and as biodiesel. We also look into how engineering of yeast can lead to improved uptake of sugars that are present in biomass hydrolyzates, and hereby allow for utilization of biomass as feedstock in the production of fuels and chemicals employing S. cerevisiae. Finally, we discuss the perspectives of how technologies from systems biology and synthetic biology can be used to advance metabolic engineering of yeast.
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| 7. |
- Skoog, Emma, 1983, et al.
(author)
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Biobased adipic acid – The challenge of developing the production host
- 2018
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In: Biotechnology Advances. - : Elsevier BV. - 0734-9750. ; 36:8, s. 2248-2263
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Research review (peer-reviewed)abstract
- Adipic acid is a platform chemical, and is the most important commercial dicarboxylic acid. It has been targeted for biochemical conversion as an alternative to present chemical production routes. From the perspective of bioeconomy, several kinds of raw material are of interest including the sugar platform (derived from starch, cellulose or hemicellulose), the lignin platform (aromatics) and the fatty acid platform (lipid derived). Two main biochemical-based production schemes may be employed: (i) direct fermentation to adipic acid, or (ii) fermentation to muconic or glucaric acid, followed by chemical hydrogenation (indirect fermentation). This review presents a comprehensive description of the metabolic pathways that could be constructed and analyzes their respective theoretical yields and metabolic constraints. The experimental yields and titers obtained so far are low, with the exception of processes based on palm oil and glycerol, which have been reported to yield up to 50 g and 68 g adipic acid/L, respectively. The challenges that remain to be addressed in order to achieve industrially relevant production levels include solving redox constraints, and identifying and/or engineering enzymes for parts of the metabolic pathways that have yet to be metabolically demonstrated. This review provides new insights into ways in which metabolic pathways can be constructed to achieve efficient adipic acid production. The production host provides the chassis to be engineered via an appropriate metabolic pathway, and should also have properties suitable for the industrial production of adipic acid. An acidic process pH is attractive to reduce the cost of downstream processing. The production host should exhibit high tolerance to complex raw material streams and high adipic acid concentrations at acidic pH.
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| 8. |
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| 9. |
- Andersson, Viktor, 1983
(author)
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Excess heat utilisation in oil refineries - CCS and algae-based biofuels
- 2016
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Doctoral thesis (other academic/artistic)abstract
- The main objective of this thesis is to investigate two different concepts for CO2 mitigation, from a system perspective, in relation to the oil refining industry: CO2 capture and storage; and algae-based biofuels. For all these processes, process integration with an oil refinery is assumed. The oil refinery sector is a major emitter of CO2 and is responsible for 9% of the industrial emissions of CO2 worldwide. Oil refineries have large amounts of unused excess heat, which can be used to satisfy the heat demands of a CO2 capture plant, a land-based algal cultivation facility, or an algae-based biofuel process. The use of this excess heat significantly reduces the cost for CO2 capture, while an economic evaluation for algae-based biofuels has not been made.Since the amount of heat available from the oil refinery´s processes increase with decreasing temperature in the stripper reboiler, it was investigated how much heat was available at different temperatures. It was also investigated how the decreased temperature would affect the heat demand of CO2 capture processes that use MEA or ammonia as the absorbent. The findings show that it is possible to capture more CO2 using excess heat when the temperature in the stripper reboiler is decreased. For the MEA process, the lower limit of the temperature interval investigated showed the maximum CO2 capture rate, while the ammonia process benefitted from a lower temperature than the standard temperature but showed maximal CO2 capture rate above the lower limit. These results are valid only when using excess heat to satisfy the entire heat demand. At the case study refinery, the available excess heat could satisfy between 28% and 50% of the heat demand of the MEA process when treating the flue gases from all chimneys, depending on the temperature in the stripper reboiler. This utilisation of excess heat represents a way to reduce significantly the costs for CCS in an oil refinery. Land-based cultivation of algae proved to be unsuitable for the utilisation of excess heat. Since the cultivation pond is exposed to wind, rain, and cold, the heat demand fluctuates strongly over the year, making the pond an unstable recipient of the excess heat.Three types of biofuel processes based on microalgae and macroalgae were investigated with respect to integration with the oil refinery. For the algae-based biofuel processes, heat integration and material integration combined to increase the efficiency of the system. When two different build margin technologies (with different CO2 emission factors) are employed for electricity production, macroalgae-based biofuel production appears to be the more robust process from the perspective of CO2 due to the lower electricity demands of the algal cultivation and harvesting phases.
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| 10. |
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| 11. |
- Anasontzis, George E, 1980
(author)
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Biomass modifying enzymes: From discovery to application
- 2012
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In: Oral presentation at the Chalmers Life Science AoA conference.
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Conference paper (other academic/artistic)abstract
- It has now been realized that the road towards the bio-based economy is a one-way street, leaving gradually the oil-based technology and driving slowly towards a more sustainable society. The current non-biodegradable hydrocarbon fuels and plastics will be replaced by new products which will derive from natural and renewable resources. The synthesis of such biofuels and biochemicals is still challenged by the difficulties to cost efficiently degrade lignocellulosic material to fermentable sugars or to isolate the intact polymers. Biomass degrading and modifying enzymes play an integral role both in the separation of the polymers from the wood network, as well as in their subsequent modification, prior to further product development.Our group interests focus on all levels of applied enzyme research of biomass acting enzymes: Discovery, assay development, production and application. Relevant examples will be provided: What is our strategy for discovering novel microorganisms and enzymes from the tropical forests and grasslands of Vietnam? How do we design novel real-world assays for enzyme activity determination? Which are the bottlenecks in the enzymatic cellulose hydrolysis? How enzymes can be used to produce high added value compounds from biomass?
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| 12. |
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| 13. |
- Gullfot, Fredrika, 1967-
(author)
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Synthesis of xyloglucan oligo- and polysaccharides with glycosynthase technology
- 2009
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Licentiate thesis (other academic/artistic)abstract
- Xyloglucans are polysaccharides found as storage polymers in seeds and tubers, and as cross-linking glycans in the cell wall of plants. Their structure is complex with intricate branching patterns, which contribute to the physical properties of the polysaccharide including its binding to and interaction with other glycans such as cellulose. Xyloglucan is widely used in bulk quantities in the food, textile and paper making industries. With an increasing interest in technically more advanced applications of xyloglucan, such as novel biocomposites, there is a need to understand and control the properties and interactions of xyloglucan with other compounds, to decipher the relationship between xyloglucan structure and function, and in particular the effect of different branching patterns. However, due to the structural heterogeneity of the polysaccharide as obtained from natural sources, relevant studies have not been possible to perform in practise. This fact has stimulated an interest in synthetic methods to obtain xyloglucan mimics and analogs with well-defined structure and decoration patterns. Glycosynthases are hydrolytically inactive mutant glycosidases that catalyse the formation of glycosidic linkages between glycosyl fluoride donors and glycoside acceptors. Since its first conception in 1998, the technology is emerging as a useful tool in the synthesis of large, complex polysaccharides. This thesis presents the generation and characterisation of glycosynthases based on xyloglucanase scaffolds for the synthesis of well-defined homogenous xyloglucan oligo- and polysaccharides with regular substitution patterns.
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| 14. |
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| 15. |
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| 16. |
- Steinhagen, Sophie, et al.
(author)
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Harvest time can affect the optimal yield and quality of sea lettuce (Ulva fenestrata) in a sustainable sea-based cultivation : Seasonal Cultivation of Ulva fenestrata
- 2022
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In: Frontiers in Marine Science. - : Frontiers Media SA. - 2296-7745. ; 9
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Journal article (peer-reviewed)abstract
- Seaweed biomass is a renewable resource with multiple applications. Sea-based cultivation of seaweeds can provide high biomass yields, low construction, operation, and maintenance costs and could offer an environmentally and economically sustainable alternative to land-based cultivations. The biochemical profile of sea-grown biomass depends on seasonal variation in environmental factors, and the optimization of harvest time is important for the quality of the produced biomass. To identify optimal harvest times of Swedish sea-based cultivated sea lettuce (Ulva fenestrata), this study monitored biomass yield, morphology, chemical composition, fertility, and biofouling at five different harvesting times in April - June 2020. The highest biomass yields (approx. 1.2 kg fw [m rope]-1) were observed in late spring (May). The number and size of holes in the thalli and the amount of fertile and fouled tissue increased with prolonged growth season, which together led to a significant decline in both biomass yield and quality during summer (June). Early spring (April) conditions were optimal for obtaining high fatty acid, protein, biochar, phenolic, and pigment contents in the biomass, whereas carbohydrate and ash content, as well as essential and non-essential elements, increased later in the growth season. Our study results show that the optimal harvest time of sea-based cultivated U. fenestrata depends on the downstream application of the biomass and must be carefully selected to balance yield, quality, and desired biochemical contents to maximize the output of future sea-based algal cultivations in the European Northern Hemisphere.
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| 17. |
- Iseri, Emre
(author)
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Microfluidic Compartmentalization for Smart Materials, Medical Diagnostics and Cell Therapy
- 2022
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Doctoral thesis (other academic/artistic)abstract
- The organisation of fluids in small compartments is ubiquitous in nature, such as in the cellular composition of all life. This work explores several engineering avenues where microscale fluid compartmentalization can bring novel material properties or novel functionality in life sciences or medicine. Here, we introduce four unique compartmentalization methods: 1) 3D fluid self-organisation in microscaffolds (FLUID3EAMS), 2) 2D microcapillary arrays on a dipstick (Digital Dipstick), 3) a sliding microfluidic platform with cross-flow (Slip-X-Chip), and 4) compartmentalization by cutting of soft solid matter (Solidify & Cut). These methods were used in a wide range of applications. Within the area of smart materials, we applied FLUID3EAMS to synthesize materials with temperature-tuneable permeability and surface energy and to establish, in a well-controlled fashion, tissue-like materials in the form of 3D droplet interface bilayer networks. Solidify & Cut was used to form soft composites with a new type of magnetic behaviour, rotation-induced ferromagnetism, that allows easy reprogramming of the magnetization of magnetopolymers. Within the area of medical diagnostics, we applied Digital Dipstick to perform rapid digital bacterial culture in a dipstick format and obtained clinically relevant diagnostic results on samples from patients with a urinary tract infection. Furthermore, Slip-X-Chip enables particle concentration and washing as new functions in sliding microfluidic platforms, which significantly expands their potential application area. Finally, within the area of cell therapy, we explored the microencapsulation of high concentrations of therapeutic cells and presented a novel technique to fabricate core-shell microcapsules by exploiting the superior material properties of spider silk membranes.
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| 18. |
- Ask, Magnus, 1983
(author)
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Towards More Robust Saccharomyces cerevisiae Strains for Lignocellulosic Bioethanol Production: Lessons from process concepts and physiological investigations
- 2013
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Doctoral thesis (other academic/artistic)abstract
- Dwindling oil reserves and the negative impacts of fossil fuels on the environment call for more sustainable energy sources. First-generation bioethanol produced from sugar cane and corn has met some of these needs, but it competes with the food supply for raw materials. Lignocellulosic biomass is an abundant non-edible raw material that can be converted to ethanol using the yeast Saccharomyces cerevisiae. However, due to the inherent recalcitrance to degradation of lignocellulosic raw materials, harsh pretreatment methods must be used to liberate fermentable sugars, resulting in the release of compounds such as acetic acid, furan aldehydes and phenolics, that inhibit yeast metabolism. This thesis research aimed to identify bottlenecks in terms of inhibitory compounds related to ethanol production from two lignocellulosic raw materials, Arundo donax and spruce, and furthermore to harness the physiological responses to these inhibitors to engineer more robust yeast strains. A comparative study of separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) revealed that acetic acid limits xylose utilization in pretreated Arundo donax, whereas the furan aldehydes furfural and 5-hydroxymethyl-2-furaldehyde (HMF) were hypothesized to be key inhibitors in pretreated spruce. The impacts of furfural and HMF on the redox and energy metabolism of S. cerevisiae were studied in detail in chemostat and batch cultivations. After adding the inhibitors to the feed medium of chemostat cultivations, the intracellular levels of NADH, NADPH, and ATP were found to decrease by 40, 75, and 19%, respectively, suggesting that furan aldehydes drain the cells of reducing power. A strong effect on redox metabolism was also observed after pulsing furfural and HMF in the xylose consumption phase in batch cultures. The drainage of reducing power was also observed in a genome-wide study of transcription that found that genes related to NADPH-requiring processes, such as nitrogen and sulphur assimilation, were significantly induced. The redox metabolism was engineered by overproducing the protective metabolite and antioxidant glutathione. Strains with an increased intracellular level of reduced glutathione were found to sustain ethanol production for longer duration in SSF of pretreated spruce, yielding 70% more ethanol than did the wild type strain.
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| 19. |
- Ylitervo, Päivi
(author)
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Concepts for improving ethanol productivity from lignocellulosic materials : encapsulated yeast and membrane bioreactors
- 2014
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Doctoral thesis (other academic/artistic)abstract
- Lignocellulosic biomass is a potential feedstock for production of sugars, which can be fermented into ethanol. The work presented in this thesis proposes some solutions to overcome problems with suboptimal process performance due to elevated cultivation temperatures and inhibitors present during ethanol production from lignocellulosic materials. In particular, continuous processes operated at high dilution rates with high sugar utilisation are attractive for ethanol fermentation, as this can result in higher ethanol productivity. Both encapsulation and membrane bioreactors were studied and developed to achieve rapid fermentation at high yeast cell density. My studies showed that encapsulated yeast is more thermotolerant than suspended yeast. The encapsulated yeast could successfully ferment all glucose during five consecutive batches, 12 h each at 42 °C. In contrast, freely suspended yeast was inactivated already in the second or third batch. One problem with encapsulation is, however, the mechanical robustness of the capsule membrane. If the capsules are exposed to e.g. high shear forces, the capsule membrane may break. Therefore, a method was developed to produce more robust capsules by treating alginate-chitosan-alginate (ACA) capsules with 3-aminopropyltriethoxysilane (APTES) to get polysiloxane-ACA capsules. Of the ACA-capsules treated with 1.5% APTES, only 0–2% of the capsules broke, while 25% of the untreated capsules ruptured within 6 h in a shear test. In this thesis membrane bioreactors (MBR), using either a cross-flow or a submerged membrane, could successfully be applied to retain the yeast inside the reactor. The cross-flow membrane was operated at a dilution rate of 0.5 h-1 whereas the submerged membrane was tested at several dilution rates, from 0.2 up to 0.8 h-1. Cultivations at high cell densities demonstrated an efficient in situ detoxification of very high furfural levels of up to 17 g L-1 in the feed medium when using a MBR. The maximum yeast density achieved in the MBR was more than 200 g L-1. Additionally, ethanol fermentation of nondetoxified spruce hydrolysate was possible at a high feeding rate of 0.8 h-1 by applying a submerged membrane bioreactor, resulting in ethanol productivities of up to 8 g L-1 h-1. In conclusion, this study suggests methods for rapid continuous ethanol production even at stressful elevated cultivation temperatures or inhibitory conditions by using encapsulation or membrane bioreactors and high cell density cultivations.
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| 20. |
- Jansson, Ronnie, et al.
(author)
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Functionalized silk assembled from a recombinant spider silk fusion protein (Z-4RepCT) produced in the methylotrophic yeast Pichia pastoris
- 2016
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In: Biotechnology Journal. - : Wiley-VCH Verlagsgesellschaft. - 1860-6768 .- 1860-7314. ; 11:5, s. 687-699
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Journal article (peer-reviewed)abstract
- Functional biological materials are a growing research area with potential applicability in medicine and biotechnology. Using genetic engineering, the possibility to introduce additional functions into spider silk-based materials has been realized. Recently, a recombinant spider silk fusion protein, Z-4RepCT, was produced intracellularly in Escherichia coli and could after purification self-assemble into silk-like fibers with ability to bind antibodies via the IgG-binding Z domain. In this study, the use of the methylotrophic yeast Pichia pastoris for production of Z-4RepCT has been investigated. Temperature, pH and production time were influencing the amount of soluble Z-4RepCT retrieved from the extracellular fraction. Purification of secreted Z-4RepCT resulted in a mixture of full-length and degraded silk proteins that failed to self-assemble into fibers. A position in the C-terminal domain of 4RepCT was identified as being subjected to proteolytic cleavage by proteases in the Pichia culture supernatant. Moreover, the C-terminal domain was subjected to glycosylation during production in P. pastoris. These observed alterations of the CT domain are suggested to contribute to the failure in fiber assembly. As alternative approach, Z-4RepCT retrieved from the intracellular fraction, which was less degraded, was used and shown to retain ability to assemble into silk-like fibers after enzymatic deglycosylation.
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| 21. |
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| 22. |
- Bergman, Alexandra Linda, 1985, et al.
(author)
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Heterologous phosphoketolase expression redirects flux towards acetate, perturbs sugar phosphate pools and increases respiratory demand in Saccharomyces cerevisiae
- 2019
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In: Microbial Cell Factories. - : Springer Science and Business Media LLC. - 1475-2859. ; 18:1
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Journal article (peer-reviewed)abstract
- Introduction: Phosphoketolases (Xfpk) are a non-native group of enzymes in yeast, which can be expressed in combination with other metabolic enzymes to positively influence the yield of acetyl-CoA derived products by reducing carbon losses in the form of CO2. In this study, a yeast strain expressing Xfpk from Bifidobacterium breve, which was previously found to have a growth defect and to increase acetate production, was characterized. Results: Xfpk-expression was found to increase respiration and reduce biomass yield during glucose consumption in batch and chemostat cultivations. By cultivating yeast with or without Xfpk in bioreactors at different pHs, we show that certain aspects of the negative growth effects coupled with Xfpk-expression are likely to be explained by proton decoupling. At low pH, this manifests as a reduction in biomass yield and growth rate in the ethanol phase. Secondly, we show that intracellular sugar phosphate pools are significantly altered in the Xfpk-expressing strain. In particular a decrease of the substrates xylulose-5-phosphate and fructose-6-phosphate was detected (26% and 74% of control levels) together with an increase of the products glyceraldehyde-3-phosphate and erythrose-4-phosphate (208% and 542% of control levels), clearly verifying in vivo Xfpk enzymatic activity. Lastly, RNAseq analysis shows that Xfpk expression increases transcription of genes related to the glyoxylate cycle, the TCA cycle and respiration, while expression of genes related to ethanol and acetate formation is reduced. The physiological and transcriptional changes clearly demonstrate that a heterologous phosphoketolase flux in combination with endogenous hydrolysis of acetyl-phosphate to acetate increases the cellular demand for acetate assimilation and respiratory ATP-generation, leading to carbon losses. Conclusion: Our study shows that expression of Xfpk in yeast diverts a relatively small part of its glycolytic flux towards acetate formation, which has a significant impact on intracellular sugar phosphate levels and on cell energetics. The elevated acetate flux increases the ATP-requirement for ion homeostasis and need for respiratory assimilation, which leads to an increased production of CO2. A majority of the negative growth effects coupled to Xfpk expression could likely be counteracted by preventing acetate accumulation via direct channeling of acetyl-phosphate towards acetyl-CoA.
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| 23. |
- Westman, Johan, 1983, et al.
(author)
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Current progress in high cell density yeast bioprocesses for bioethanol production
- 2015
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In: Biotechnology journal. - : Wiley. - 1860-6768 .- 1860-7314. ; 10:8, s. 1185-1195
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Research review (peer-reviewed)abstract
- High capital costs and low reaction rates are major challenges for establishment of fermentation-based production systems in the bioeconomy. Using high cell density cultures is an efficient way to increase the volumetric productivity of fermentation processes, thereby enabling faster and more robust processes and use of smaller reactors. In this review, we summarize recent progress in the application of high cell density yeast bioprocesses for first and second generation bioethanol production. High biomass concentrations obtained by retention of yeast cells in the reactor enables easier cell reuse, simplified product recovery and higher dilution rates in continuous processes. High local cell density cultures, in the form of encapsulated or strongly flocculating yeast, furthermore obtain increased tolerance to convertible fermentation inhibitors and utilize glucose and other sugars simultaneously, thereby overcoming two additional hurdles for second generation bioethanol production. These effects are caused by local concentration gradients due to diffusion limitations and conversion of inhibitors and sugars by the cells, which lead to low local concentrations of inhibitors and glucose. Quorum sensing may also contribute to the increased stress tolerance. Recent developments indicate that high cell density methodology, with emphasis on high local cell density, offers significant advantages for sustainable second generation bioethanol production.
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| 24. |
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| 25. |
- Wang, Shule, 1994-, et al.
(author)
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Effect of H2 as Pyrolytic Agent on the Product Distribution during Catalytic Fast Pyrolysis of Biomass Using Zeolites
- 2018
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In: Energy & Fuels. - : American Chemical Society (ACS). - 0887-0624 .- 1520-5029. ; 32:8, s. 8530-8536
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Journal article (peer-reviewed)abstract
- Bio-oil generated from catalytic fast pyrolysis or hydrotreating processes represents one of the most promising alternatives to liquid fossil fuels. The use of H2 as carrier gas in the pyrolysis of biomass requires further research to study the catalytic fast pyrolysis reactions in the case of using reactive atmosphere. In this work, pyrolysis experiments with lignocellulosic biomass have been performed in a fixed bed reactor in H2 and N2 atmospheres with/without HZSM-5 additions to investigate the influence of the pyrolytic agents during fast pyrolysis of biomass and upgrading of pyrolytic vapors over a zeolitic catalyst. It was found that in a H2 atmosphere, H2 was consumed in both noncatalytic and catalytic pyrolysis processes, respectively. Higher yields of nonaqueous liquids and permanent gases are obtained in a H2 atmosphere compared to a N2 atmosphere. A catalytic pyrolysis process using HZSM-5 in a H2 atmosphere increased the production of polymer aromatic hydrocarbons and suppressed the production of monomer aromatic hydrocarbons compared to similar tests performed in a N2 atmosphere. The results show an overall increased activity of HZSM-5 in the reactive H2 atmosphere compared to a N2 atmosphere.
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| 26. |
- Rems, Lea, et al.
(author)
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Cell electrofusion using nanosecond electric pulses
- 2013
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In: Scientific Reports. - : Macmillan Publishers Ltd.. - 2045-2322. ; 3
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Journal article (peer-reviewed)abstract
- Electrofusion is an efficient method for fusing cells using short-duration high-voltage electric pulses. However, electrofusion yields are very low when fusion partner cells differ considerably in their size, since the extent of electroporation (consequently membrane fusogenic state) with conventionally used microsecond pulses depends proportionally on the cell radius. We here propose a new and innovative approach to fuse cells with shorter, nanosecond (ns) pulses. Using numerical calculations we demonstrate that ns pulses can induce selective electroporation of the contact areas between cells (i.e. the target areas), regardless of the cell size. We then confirm experimentally on B16-F1 and CHO cell lines that electrofusion of cells with either equal or different size by using ns pulses is indeed feasible. Based on our results we expect that ns pulses can improve fusion yields in electrofusion of cells with different size, such as myeloma cells and B lymphocytes in hybridoma technology.
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| 27. |
- Guo, Weijin, et al.
(author)
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Synthetic Paper Separates Plasma from Whole Blood with Low Protein Loss
- 2020
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In: Analytical Chemistry. - Washington D.C. : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 92:9, s. 6194-6199
-
Journal article (peer-reviewed)abstract
- The separation of plasma from whole blood is the first step in many diagnostic tests. Point-of-care tests often rely on integrated plasma filters, but protein retention in such filters limits their performance. Here, we investigate plasma separation on interlocked micropillar scaffolds ("synthetic paper") by the local agglutination of blood cells coupled with the capillary separation of the plasma. We separated clinically relevant volumes of plasma with high efficiency in a separation time on par with that of state of the art techniques. We investigated different covalent and non-covalent surface treatments (PEGMA, HEMA, BSA, O2 plasma) on our blood filter and their effect on protein recovery, and identified O2 plasma treatment and 7.9 μg/cm2 agglutination antibody as most suitable treatments. Using these treatments, we recovered at least 82% of the blood plasma proteins, more than with state-of-the-art filters. The simplicity of our device and the performance of our approach could enable better point-of-care tests.
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| 28. |
- Cutas, Daniela, 1978, et al.
(author)
-
Legal imperialism in the regulation of stem cell research and therapy: the problem of extraterritorial jurisdiction
- 2010
-
In: Capps BJ & Campbell AV (eds.). CONTESTED CELLS: Global Perspectives on the Stem Cell Debate. - London : Imperial College Press. - 9781848164376 ; , s. 95-119
-
Book chapter (other academic/artistic)abstract
- Countries worldwide have very different national regulations on human embryonic stem (ES) cell research, informed by a range of ethical values. Some countries find reason to extend the applicability of their regulations on such research to its citizens when they visit other countries. Extraterritorial jurisdiction has recently been identified as a potential challenge towards global regulation of ES cell research. This chapter explores the implications and impact of extraterritorial jurisdiction and global regulation of ES cell research on researchers, clinicians and national health systems, and how this may affect patients. The authors argue that it would make ethical sense for ES cell restrictive countries to extend its regulations on ES cell research beyond its borders, because, if these countries really consider embryo destruction to be objectionable on the basis on the status of the embryo, then they ought to count it morally on par with murder (and thus have a moral imperative to protect embryos from the actions of its own citizens). However, doing so could lead to a legal situation that would result in substantial harm to central values in areas besides research, such as health care, the job market, basic freedom of movement, and strategic international finance and politics. Thus, it seems that restrictive extraterritorial jurisdiction in respect to ES cell research would be deeply problematic, given that the ethical permissibility of ES cell research is characterised by deep and wide disagreement.
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| 29. |
- Franzén, Carl Johan, 1966, et al.
(author)
-
Multifeed simultaneous saccharification and fermentation enables high gravity submerged fermentation of lignocellulose.
- 2015
-
In: Recent Advances in Fermentation Technology (RAFT 11), Clearwater Beach, Florida, USA, November 8-11, 2015. Oral presentation..
-
Conference paper (other academic/artistic)abstract
- Today, second generation bioethanol production is becoming established in production plants across the world. In addition to its intrinsic value, the process can be viewed as a model process for biotechnological conversion of recalcitrant lignocellulosic raw materials to a range of chemicals and other products. So called High Gravity operation, i.e. fermentation at high solids loadings, represents continued development of the process towards higher product concentrations and productivities, and improved energy and water economy. We have employed a systematic, model-driven approach to the design of feeding schemes of solid substrate, active yeast adapted to the actual substrate, and enzymes to fed-batch simultaneous saccharification and co-fermentation (Multifeed SSCF) of steam-pretreated lignocellulosic materials in stirred tank reactors. With this approach, mixing problems were avoided even at water insoluble solids contents of 22%, leading to ethanol concentrations of 56 g/L within 72 hours of SSCF on wheat straw. Similar fermentation performance was verified in 10 m3 demonstration scale using wheat straw, and in lab scale on birch and spruce, using several yeast strains. The yeast was propagated in the liquid fraction obtained by press filtration of the pretreated slurry. Yet, even with such preadaptation and repeated addition of fresh cells, the viability in the SSCF dropped due to interactions between lignocellulose-derived inhibitors, the produced ethanol and the temperature. Decreasing the temperature from 35 to 30°C when the ethanol concentration reached 40-50 g/L resulted in rapid initial hydrolysis, maintained fermentation capacity, lower residual glucose and xylose and ethanol concentrations above 60 g/L.
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| 30. |
- Nickel, David, 1990, et al.
(author)
-
Uncertainty analysis as a tool to consistently evaluate lignocellulosic bioethanol processes at different system scales
- 2018
-
Conference paper (other academic/artistic)abstract
- Lignocellulosic processes are highly prone to batch-to batch variability, e.g. of raw materials and enzyme activities. This variability can be propagated throughout system scales during process development and optimization, influencing the outputs of bioreaction models, techno-economic analyses and life cycle assessments. As these outputs are the main decision variables for designing and developing lignocellulose-based processes, tools are required to evaluate the influences of process variation at different system scales. Uncertainty analysis quantifies the effects of model input variations on model outputs. It is an effective tool to consistently propagate process variation throughout scales and analyse its influence on model outputs. As an example, we use a model describing multi-feed simultaneous saccharification and co-fermentation (SSCF) of wheat straw. During the process enzymes hydrolyse the lignocellulosic material to release glucose which can be converted by microorganisms into ethanol. To investigate the impact of batch-to-batch variability in enzyme cocktails, we collected literature data on the enzymatic activity of Cellic CTec2. Retrieved data were propagated in models at bioreactor, techno-economic analysis and life cycle assessment scale. We show how uncertainty analysis can be used to guide process development by comparing different modes of operation. The method can identify economically feasible process ranges with low environmental impact while increasing the robustness of bioprocesses with high variation in raw material inputs. Furthermore, uncertainty analysis could help to identify relevant parameters to choose as response variables in experimental designs.
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| 31. |
- Wang, Ruifei, 1985, et al.
(author)
-
Model-based optimization and scale-up of multi-feed simultaneous saccharification and co-fermentation of steam pre-treated lignocellulose enables high gravity ethanol production.
- 2016
-
In: Biotechnology for Biofuels. - : Springer Science and Business Media LLC. - 1754-6834 .- 1754-6834. ; 9:1, s. 88-
-
Journal article (peer-reviewed)abstract
- High content of water-insoluble solids (WIS) is required for simultaneous saccharification and co-fermentation (SSCF) operations to reach the high ethanol concentrations that meet the techno-economic requirements of industrial-scale production. The fundamental challenges of such processes are related to the high viscosity and inhibitor contents of the medium. Poor mass transfer and inhibition of the yeast lead to decreased ethanol yield, titre and productivity. In the present work, high-solid SSCF of pre-treated wheat straw was carried out by multi-feed SSCF which is a fed-batch process with additions of substrate, enzymes and cells, integrated with yeast propagation and adaptation on the pre-treatment liquor. The combined feeding strategies were systematically compared and optimized using experiments and simulations.
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| 32. |
- Ylitervo, Päivi, et al.
(author)
-
Continuous Ethanol Production with a Membrane Bioreactor at High Acetic Acid Concentrations
- 2014
-
In: Membranes. - : MDPI. - 2077-0375. ; 4:3, s. 372-387
-
Journal article (peer-reviewed)abstract
- The release of inhibitory concentrations of acetic acid from lignocellulosic raw materials during hydrolysis is one of the main concerns for 2nd generation ethanol production. The undissociated form of acetic acid can enter the cell by diffusion through the plasma membrane and trigger several toxic effects, such as uncoupling and lowered intracellular pH. The effect of acetic acid on the ethanol production was investigated in continuous cultivations by adding medium containing 2.5 to 20.0 g•L−1 acetic acid at pH 5.0, at a dilution rate of 0.5 h−1. The cultivations were performed at both high (~25 g•L−1) and very high (100–200 g•L−1) yeast concentration by retaining the yeast cells inside the reactor by a cross-flow membrane in a membrane bioreactor. The yeast was able to steadily produce ethanol from 25 g•L−1 sucrose, at volumetric rates of 5–6 g•L−1•h−1 at acetic acid concentrations up to 15.0 g•L−1. However, the yeast continued to produce ethanol also at a concentration of 20 g•L−1 acetic acid but at a declining rate. The study thereby demonstrates the great potential of the membrane bioreactor for improving the robustness of the ethanol production based on lignocellulosic raw materials.
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| 33. |
- Punzi, Marisa, et al.
(author)
-
Combined anaerobic-ozonation process for treatment of textile wastewater : Removal of acute toxicity and mutagenicity
- 2015
-
In: Journal of Hazardous Materials. - : Elsevier BV. - 0304-3894 .- 1873-3336. ; 292, s. 52-60
-
Journal article (peer-reviewed)abstract
- A novel set up composed of an anaerobic biofilm reactor followed by ozonation was used for treatment of artificial and real textile effluents containing azo dyes. The biological treatment efficiently removed chemical oxygen demand and color. Ozonation further reduced the organic content of the effluents and was very important for the degradation of aromatic compounds, as shown by the reduction of UV absorbance. The acute toxicity toward Vibrio fischeri and the shrimp Artemia salina increased after the biological treatment. No toxicity was detected after ozonation with the exception of the synthetic effluent containing the highest concentration, 1. g/l, of the azo dye Remazol Red. Both untreated and biologically treated textile effluents were found to have mutagenic effects. The mutagenicity increased even further after 1. min of ozonation. No mutagenicity was however detected in the effluents subjected to longer exposure to ozone. The results of this study suggest that the use of ozonation as short post-treatment after a biological process can be beneficial for the degradation of recalcitrant compounds and the removal of toxicity of textile wastewater. However, monitoring of toxicity and especially mutagenicity is crucial and should always be used to assess the success of a treatment strategy. © 2015 Elsevier B.V.
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| 34. |
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| 35. |
- Nickel, David, 1990, et al.
(author)
-
Multi-scale uncertainty analysis – A tool to systematically consider variability in lignocellulosic bioethanol processes
- 2018
-
Conference paper (other academic/artistic)abstract
- Bioethanol production processes from lignocellulosic raw materials are highly prone to batch-to-batch variations. For example, raw material compositions and enzymatic activities required to release fermentable sugars from lignocellulose vary significantly between batches. To develop lignocellulosic biofuel processes and evaluate their performance regarding economics and sustainability consistently, tools are required to cope with this variability. In this presentation we will propose a multi-scale uncertainty analysis strategy to propagate input variability throughout system scales. In a first step, we use meta-data obtained from literature to define uncertainties in the process inputs. Utilizing these meta-data, uncertainty analysis is performed on a macro-kinetic model by sampling from the defined uncertain input space. The results of this uncertainty analysis are transferred to process simulations to analyze the impact of input uncertainties on the process mass- and energy balances, and on the economics of building this type of bioprocess. The generated data from process simulations (mass flows, energy integration, and economic data) are in the next step extracted and used as input to an environmental impact assessment of the process. This is done whilst keeping the simulation and systems modeling parameters constant, thus the input variability is propagated throughout the different system scales. The data generated in this integrated approach will then be compared with the variations and uncertainties observed with relevance to the estimated parameters in the process simulation and environmental impact assessment. Based on this consistent strategy, we can analyze the impact of input variability from different system perspectives, identify important bottlenecks for development, and suggest robust and sustainable process designs for different conditions and under given uncertainties. In a case study we demonstrate how integrated kinetic modeling (in Matlab), process simulation (in SuperPro Designer), and environmental impact assessment together with statistical analysis can be used for assessing how variability in enzymatic activities in bioethanol production can be propagated throughout system scales. A macro-kinetic model is used to describe the enzymatic breakdown of lignocellulose-derived polysaccharides into fermentable sugars (saccharification) and the simultaneous fermentation to bioethanol. We discuss the integration of the simulation results of the macro-kinetic model into the flowsheeting software for mass and energy balance generation, and then further on to assess environmental impacts of the process. We will evaluate different process designs regarding their robustness towards input variability. Finally, we also show how propagated uncertainties at different system scales can be integrated to design experiments at laboratory scale so that these focus on the most important parameters for developing robust kinetic models, and include the parameters that are most important for sustainable design of processes and value chains.
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| 36. |
- Nickel, David, 1990
(author)
-
Process development for platform chemical production from agricultural and forestry residues
- 2021
-
Doctoral thesis (other academic/artistic)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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| 37. |
- Skvaril, Jan, 1982-, et al.
(author)
-
Applications of near-infrared spectroscopy (NIRS) in biomass energy conversion processes : A review
- 2017
-
In: Applied spectroscopy reviews (Softcover ed.). - : Informa UK Limited. - 0570-4928 .- 1520-569X. ; 52:8, s. 675-728
-
Research review (peer-reviewed)abstract
- Biomass used in energy conversion processes is typically characterized by high variability, making its utilization challenging. Therefore, there is a need for a fast and non-destructive method to determine feedstock/product properties and directly monitor process reactors. The near-infrared spectroscopy (NIRS) technique together with advanced data analysis methods offers a possible solution. This review focuses on the introduction of the NIRS method and its recent applications to physical, thermochemical, biochemical and physiochemical biomass conversion processes represented mainly by pelleting, combustion, gasification, pyrolysis, as well as biogas, bioethanol, and biodiesel production. NIRS has been proven to be a reliable and inexpensive method with a great potential for use in process optimization, advanced control, or product quality assurance.
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| 38. |
- Forsgren, Johan, et al.
(author)
-
Co-loading of bisphosphonates and antibiotics to a biomimetic hydroxyapatite coating.
- 2011
-
In: Biotechnology letters. - : Springer Science and Business Media LLC. - 1573-6776 .- 0141-5492. ; 33:6, s. 1265-8
-
Journal article (peer-reviewed)abstract
- We have incorporated bisphosphonates and antibiotics simultaneously into a biomimetic hydroxyapatite implant coating aiming to use the interaction between drug-molecules and hydroxyapatite to enable local release of the two different substances to obtain a dual biological effect. A sustained release over for 43h of antibiotics (cephalothin) was achieved without negative interference from the presence of the bisphosphonate (clodronate) which, in turn, successfully bonded to the coating surface. To our knowledge, this is the first study that indicates the possibility to simultaneously incorporate both antibiotics and bisphosphonates to an implant coating, a strategy that is believed to improve implant stability and reduce implant-related infections.
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| 39. |
- Koskela, Salla, et al.
(author)
-
Hemicellulose content affects the properties of cellulose nanofibrils produced from softwood pulp fibres by LPMO
- 2022
-
In: Green Chemistry. - : Royal Society of Chemistry (RSC). - 1463-9262 .- 1463-9270.
-
Journal article (peer-reviewed)abstract
- Lytic polysaccharide monooxygenase (LPMO)-catalysed oxidation of cellulose has emerged as a green alternative to chemical modifications in the production of cellulose nanofibrils (CNFs) from wood pulp fibres. The effect of the hemicellulose content of the starting pulp fibres on the oxidation capabilities of cellulose-active LPMO is important and has not been investigated previously. In this study, the production of LPMO-oxidised CNFs was evaluated on two commercial softwood pulp fibres with different hemicellulose contents. Thin and colloidally stable CNFs were readily obtained from kraft pulp with a hemicellulose content of 16%. The preserved hemicellulose fraction in the kraft pulp enhanced the access of LPMO into the fibre cell wall, enabling the production of homogeneous CNFs with a thin width of 3.7 ± 1.7 nm. By contrast, the LPMO-oxidised dissolving pulp with a lower hemicellulose content of 4% could only be partially disintegrated into thin CNFs, leaving a large amount of cellulose microfibril aggregates with widths of around 50 to 100 nm. CNFs disintegrated from the LPMO-oxidised kraft pulp could be processed into nanopapers with excellent properties including an optical transmittance of 86%, tensile strength of 260 MPa, and Young's modulus of 16.9 GPa. Such CNFs also showed acid-triggered nanofibril gelation owing to the introduced carboxyl groups on cellulose microfibril surfaces. These results indicate that the inherent hemicelluloses present in the wood cell wall are essential for LPMO-mediated CNF production from wood pulp fibres.
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| 40. |
- Zambrano, Jesús, et al.
(author)
-
Optimal steady-state design of zone volumes of bioreactors with Monod growth kinetics
- 2015
-
In: Biochemical Engineering Journal. - : Elsevier BV. - 1369-703X .- 1873-295X. ; 100, s. 59-66
-
Journal article (peer-reviewed)abstract
- This paper deals with steady-state analysis and design of bioreactors consisting of a number of completely stirred tank reactors (CSTRs) in series. The study is confined to one consumed (substrate) and one consuming constituent (biomass). The specific microbial growth rate is assumed to be described by Monod kinetics. The death of biomass is assumed to be negligible. Two optimal design problems for a large number of CSTRs in series are studied: to minimize the effluent substrate concentration for a given total volume, and to minimize the total volume for a given effluent substrate concentration. As an appealing alternative to solve these problems numerically, it is proposed to consider the asymptotic case where the number of CSTRs tends to infinity. This is shown to correspond to one CSTR in series with a plug flow reactor (PFR). A CSTR with a sufficient large volume is needed to avoid wash-out of the biomass. The main result is that both design problems for the CSTR + PFR configuration have the same solution with respect to the optimal volume of the CSTR, which is given as an explicit function of the incoming substrate concentration, the volumetric flow rate and the coefficients of the Monod growth rate function. Numerical results indicate that the plug flow approach may be used as a feasible design procedure even for a reasonably low number of CSTRs in series.
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| 41. |
- Martinez Avila, Hector, 1985
(author)
-
Biofabrication, Biomechanics and Biocompatibility of Nanocellulose-based Scaffolds for Auricular Cartilage Regeneration
- 2015
-
Doctoral thesis (other academic/artistic)abstract
- In about 2:10,000 births the external part of the ear, the auricle, is severely malformed or absent. Furthermore, tumors and trauma can cause defects to the auricle. For patients with dysplasia of the auricle, and especially for children, an inconspicuous outer appearance with life-like auricles is important for their psychological and emotional well being as well as their psycho-social development. Auricular reconstruction remains a great challenge due to the complexity of surgical reconstruction using rib cartilage. Despite the advances in stem cell technology and biomaterials, auricular cartilage tissue engineering (TE) is still in an early stage of development due to critical requirements demanding appropriate mechanical properties and shape stability of the tissue-engineered construct. This thesis has focused on developing patient-specific tissue-engineered auricles for one-step surgery using a novel biomaterial, bacterial nanocellulose (BNC), seeded with human nasoseptal chondrocytes (hNC) and bone marrow mononuclear cells (MNC).Biomechanical properties of human auricle cartilage were measured and used as a benchmark for tuning BNC properties. In order to meet the biomechanical requirements, a scaffold with bilayer architecture composed of a dense BNC support layer and a macroporous structure was designed. Firstly, the biocompatibility of the dense BNC layer was investigated, demonstrating a minimal foreign body response according to standards set forth in ISO 10993. Secondly, different methods to create macroporous BNC scaffolds were studied and the redifferentiation capacity of hNCs was evaluated in vitro; revealing that macroporous BNC scaffolds support cell ingrowth, proliferation and neocartilage formation. The bilayer BNC scaffold was biofabricated and tested for endotoxins and cytotoxicity before evaluating in long-term 3D culture, and subsequently in vivo for eight weeks—in an immunocompromised animal model. The results demonstrated that the non-pyrogenic and non- cytotoxic bilayer BNC scaffold offers a good mechanical stability and maintains a structural integrity, while providing a porous 3D environment that is suitable for hNCs and MNCs to produce neocartilage, in vitro and in vivo. Furthermore, patient-specific auricular BNC scaffolds with bilayer architecture were biofabricated and seeded with autologous rabbit auricular chondrocytes (rAC) for implantation in an immunocompetent rabbit model for six weeks. The results demonstrated the shape stability of the rAC-seeded scaffolds and neocartilage depositions in the immunocompetent autologous grafts. 3D bioprinting was also evaluated for biofabrication of patient-specific, chondrocyte-laden auricular constructs using a bioink composed of nanofibrillated cellulose and alginate. Bioprinted auricular constructs showed an excellent shape and size stability after in vitro culture. Moreover, this bioink supports redifferentiation of hNCs while offering excellent printability, making this a promising approach for auricular cartilage TE. Furthermore, the use of bioreactors is essential for the development of tissue-engineered cartilage in vitro. Thus, a compression bioreactor was utilized to apply dynamic mechanical stimulation to cell-seeded constructs as a means to enhance production of extracellular matrix in vitro.In this work, a potential clinical therapy for auricular reconstruction using tissue-engineered auricles is demonstrated; where BNC is proposed as a promising non-degradable biomaterial with good chemical and mechanical stability for auricular cartilage TE. Although the primary focus of this thesis is on auricular reconstruction, the methods developed are also applicable in the regeneration of other cartilage tissues such as those found in the nose, trachea, spine and articular joints.
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| 42. |
- Harrysson, Hanna, 1987, et al.
(author)
-
Strategies for Improving the Protein Yield in pH-Shift Processing of Ulva lactuca Linnaeus: Effects of Ulvan Lyases, pH-Exposure Time, and Temperature
- 2019
-
In: Acs Sustainable Chemistry & Engineering. - : American Chemical Society (ACS). - 2168-0485. ; 7:15, s. 12688-12691
-
Journal article (peer-reviewed)abstract
- Globally, there is a need for novel vegetarian protein sources. We recently showed that the pH-shift process, using alkaline protein solubilization followed by isoelectric precipitation, is an efficient way to produce extracts with high protein concentrations from Ulva lactuca (>50% on a dry matter basis). However, the total protein yield was low, and to improve this, the effects of adding ulvan lyase, preincubating the seaweed homogenate at pH 8.5 and using different protein extraction temperatures (8 degrees C, RT and 40 degrees C), were evaluated in this study. Addition of ulvan lyase reduced protein solubility but increased the precipitation. Incubation at pH 8.5, without ulvan lyase added, significantly increased both protein solubility and precipitation at 8 degrees C and RT. Temperature per se had no effect on protein solubility, while protein precipitation increased with decreasing temperature. Highest protein yield (29%) was achieved when keeping the process at 8 degrees C with a preincubation step at pH 8.5 for 1 h. By these process modifications, the yield was 3.2 times higher than achieved by the control process (9.2%).
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| 43. |
- Juul, Louise, et al.
(author)
-
Ulva fenestrata protein – comparison of three extraction methods with respect to protein yield and protein quality
- 2021
-
In: Algal Research. - : Elsevier BV. - 2211-9264. ; 60
-
Journal article (peer-reviewed)abstract
- Seaweed is gaining attention as a possible alternative and sustainable source of proteins. This study investigates three protein extraction methods and their effect on protein yield and quality when applied to Ulva fenestrata . Two of the methods included alkaline extractions (pH-shifts); one version solubilizing the proteins at pH 8.5 and one solubilizing them at pH 8.5 followed by pH 12 (pH 8.5+12). The third method was a mechanical pressing, using a double screw press. All extraction methods were followed by isoelectric precipitation to concentrate the proteins. Extraction at pH 8.5 gave the significantly highest total protein yield after the isoelectric precipitation, followed by extraction at pH 8.5+12 and lastly mechanical extraction gave the lowest yield. Proteins extracted with both alkaline methods had a significantly higher solubility at pH 7 and pH 9, compared to proteins from the mechanical pressing. There were no significant differences between the three methods in total D/L-amino acid ratio. Amino acid cross-links measured as lysinoalanine (LAL) and lanthionine (LAN) where found in significantly higher amounts in alkali-extracted proteins compared to mechanically extracted, however not to a degree that expect to compromise functional or nutritional quality. Further, no significant difference in protein in vitro digestibility was found between extraction methods. In conclusion, results indicated that protein extraction at pH 8.5 can be recommended, especially regarding total protein yield and solubility of the final protein extract.
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| 44. |
- Ekener, Elisabeth, 1963-, et al.
(author)
-
Developing Life Cycle Sustainability Assessment methodology by applying values-based sustainability weighting - Tested on biomass based and fossil transportation fuels
- 2018
-
In: Journal of Cleaner Production. - : Elsevier BV. - 0959-6526 .- 1879-1786. ; 181, s. 337-351
-
Journal article (peer-reviewed)abstract
- The production and use of transportation fuels can lead to sustainability impacts. Assessing them simultaneously in a holistic way is a challenge. This paper examines methodology for assessing the sustainability performance of products in a more integrated way, including a broad range of social impacts. Life Cycle Sustainability Assessment (LCSA) methodology is applied for this assessment. LSCA often constitutes of the integration of results from social LCA (S-LCA), environmental life cycle assessment (E-LCA) and life cycle costing (LCC). In this study, an S-LCA from an earlier project is extended with a positive social aspect, as well as refined and detailed. E-LCA and LCC results are built from LCA database and literature. Multi Criteria Decision Analysis (MCDA) methodology is applied to integrate the results from the three different assessments into an LCSA. The weighting of key sustainability dimensions in the MCDA is performed in different ways, where the sustainability dimensions are prioritized differently priority based on the assumed values of different stakeholder profiles (Egalitarian, Hierarchist, and Individualist). The developed methodology is tested on selected biomass based and fossil transportation fuels - ethanol produced from Brazilian sugarcane and US corn/maize, and petrol produced from Russian and Nigerian crude oils, where it delineates differences in sustainability performance between products assessed. The outcome in terms of relative ranking of the transportation fuel chains based on sustain ability performance differs when applying different decision-maker profiles. This result highlights and supports views that there is no one single answer regarding which of the alternatives that is most sustainable. Rather, it depends strongly upon the worldview and values held by the decision maker. A key conclusion is that sustainability assessments should pay more attention to potential differences in underlying values held by key stakeholders in relevant societal contexts. The LCSA methodology still faces challenges regarding results integration but MCDA in combination with stakeholder profiles appears to be a useful approach to build on further.
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| 45. |
- Cámara, Elena, 1985, et al.
(author)
-
Data mining of Saccharomyces cerevisiae mutants engineered for increased tolerance towards inhibitors in lignocellulosic hydrolysates
- 2022
-
In: Biotechnology Advances. - : Elsevier BV. - 0734-9750. ; 57
-
Research review (peer-reviewed)abstract
- The use of renewable plant biomass, lignocellulose, to produce biofuels and biochemicals using microbial cell factories plays a fundamental role in the future bioeconomy. The development of cell factories capable of efficiently fermenting complex biomass streams will improve the cost-effectiveness of microbial conversion processes. At present, inhibitory compounds found in hydrolysates of lignocellulosic biomass substantially influence the performance of a cell factory and the economic feasibility of lignocellulosic biofuels and chemicals. Here, we present and statistically analyze data on Saccharomyces cerevisiae mutants engineered for altered tolerance towards the most common inhibitors found in lignocellulosic hydrolysates: acetic acid, formic acid, furans, and phenolic compounds. We collected data from 7971 experiments including single overexpression or deletion of 3955 unique genes. The mutants included in the analysis had been shown to display increased or decreased tolerance to individual inhibitors or combinations of inhibitors found in lignocellulosic hydrolysates. Moreover, the data included mutants grown on synthetic hydrolysates, in which inhibitors were added at concentrations that mimicked those of lignocellulosic hydrolysates. Genetic engineering aimed at improving inhibitor or hydrolysate tolerance was shown to alter the specific growth rate or length of the lag phase, cell viability, and vitality, block fermentation, and decrease product yield. Different aspects of strain engineering aimed at improving hydrolysate tolerance, such as choice of strain and experimental set-up are discussed and put in relation to their biological relevance. While successful genetic engineering is often strain and condition dependent, we highlight the conserved role of regulators, transporters, and detoxifying enzymes in inhibitor tolerance. The compiled meta-analysis can guide future engineering attempts and aid the development of more efficient cell factories for the conversion of lignocellulosic biomass.
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| 46. |
- Ferreira, Sofia, et al.
(author)
-
Metabolic engineering strategies for butanol production in Escherichia coli
- 2020
-
In: Biotechnology and Bioengineering. - : Wiley. - 0006-3592 .- 1097-0290. ; 117:8, s. 2571-2587
-
Research review (peer-reviewed)abstract
- The global market of butanol is increasing due to its growing applications as solvent, flavoring agent, and chemical precursor of several other compounds. Recently, the superior properties of n-butanol as a biofuel over ethanol have stimulated even more interest. (Bio)butanol is natively produced together with ethanol and acetone by Clostridium species through acetone-butanol-ethanol fermentation, at noncompetitive, low titers compared to petrochemical production. Different butanol production pathways have been expressed in Escherichia coli, a more accessible host compared to Clostridium species, to improve butanol titers and rates. The bioproduction of butanol is here reviewed from a historical and theoretical perspective. All tested rational metabolic engineering strategies in E. coli to increase butanol titers are reviewed: manipulation of central carbon metabolism, elimination of competing pathways, cofactor balancing, development of new pathways, expression of homologous enzymes, consumption of different substrates, and molecular biology strategies. The progress in the field of metabolic modeling and pathway generation algorithms and their potential application to butanol production are also summarized here. The main goals are to gather all the strategies, evaluate the respective progress obtained, identify, and exploit the outstanding challenges.
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| 47. |
- Huang, Mingtao, 1984, et al.
(author)
-
Engineering the protein secretory pathway of Saccharomyces cerevisiae enables improved protein production
- 2018
-
In: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 115:47, s. E11025-E11032
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Journal article (peer-reviewed)abstract
- Baker’s yeast Saccharomyces cerevisiae is one of the most important and widely used cell factories for recombinant protein production. Many strategies have been applied to engineer this yeast for improving its protein production capacity, but productivity is still relatively low, and with increasing market demand, it is important to identify new gene targets, especially targets that have synergistic effects with previously identified targets. Despite improved protein production, previous studies rarely focused on processes associated with intracellular protein retention. Here we identified genetic modifications involved in the secretory and trafficking pathways, the histone deacetylase complex, and carbohydrate metabolic processes as targets for improving protein secretion in yeast. Especially modifications on the endosome-to-Golgi trafficking was found to effectively reduce protein retention besides increasing protein secretion. Through combinatorial genetic manipulations of several of the newly identified gene targets, we enhanced the protein production capacity of yeast by more than fivefold, and the best engineered strains could produce 2.5 g/L of a fungal α-amylase with less than 10% of the recombinant protein retained within the cells, using fed-batch cultivation.
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| 48. |
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Systems Perspectives on Biorefineries 2012
- 2012
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Editorial collection (peer-reviewed)abstract
- Replacing fossil fuels with biomass for the production of energy carriers, materials and specialty chemicalsis a challenge that now confronts humanity. In which applications shall we use limited resourcesof biomass? How can biomass be refined into the products we want? What is an optimal design of abiorefinery? How is the most advantageous portfolio of policy instruments designed to realise the biorefineriesof the future?There is not one final answer to these questions. However, different systems studies can provide us withcomplementary pieces of the puzzle. These can be valuable by themselves, or be brought together into alarger and more complex picture. Systems perspectives on Biorefineries 2012 contains nine chapters thataddress different topics related to the immensely important issue of how the world’s biomass resourcescan, or should, be converted into the goods we need and desire. The book is far from complete, but it is acontribution and a start...
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| 49. |
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| 50. |
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