| 1. |
- Anasontzis, George E, 1980
(författare)
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Biomass modifying enzymes: From discovery to application
- 2012
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Ingår i: Oral presentation at the Chalmers Life Science AoA conference.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)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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| 2. |
- Ask, Magnus, 1983
(författare)
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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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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)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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| 3. |
- Ylitervo, Päivi
(författare)
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Concepts for improving ethanol productivity from lignocellulosic materials : encapsulated yeast and membrane bioreactors
- 2014
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)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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| 4. |
- Ylitervo, Päivi, et al.
(författare)
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Continuous Ethanol Production with a Membrane Bioreactor at High Acetic Acid Concentrations
- 2014
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Ingår i: Membranes. - : MDPI. - 2077-0375. ; 4:3, s. 372-387
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Tidskriftsartikel (refereegranskat)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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| 5. |
- Olofsson, Martin, 1975-, et al.
(författare)
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Combined Effects of Nitrogen Concentration and Seasonal Changes on the Production of Lipids in Nannochloropsis oculata
- 2014
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Ingår i: Marine Drugs. - Basel, Switzerland : MDPI AG. - 1660-3397 .- 1660-3397. ; 12:4, s. 1891-1910
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Tidskriftsartikel (refereegranskat)abstract
- Instead of sole nutrient starvation to boost algal lipid production, we addressed nutrient limitation at two different seasons (autumn and spring) during outdoor cultivation in flat panel photobioreactors. Lipid accumulation, biomass and lipid productivity and changes in fatty acid composition of Nannochloropsis oculata were investigated under nitrogen (N) limitation (nitrate:phosphate N:P 5, N:P 2.5 molar ratio). N. oculata was able to maintain a high biomass productivity under N-limitation compared to N-sufficiency (N:P 20) at both seasons, which in spring resulted in nearly double lipid productivity under N-limited conditions (0.21 g L−1 day−1) compared to N-sufficiency (0.11 g L−1 day−1). Saturated and monounsaturated fatty acids increased from 76% to nearly 90% of total fatty acids in N-limited cultures. Higher biomass and lipid productivity in spring could, partly, be explained by higher irradiance, partly by greater harvesting rate (~30%). Our results indicate the potential for the production of algal high value products (i.e., polyunsaturated fatty acids) during both N-sufficiency and N-limitation. To meet the sustainability challenges of algal biomass production, we propose a dual-system process: Closed photobioreactors producing biomass for high value products and inoculum for larger raceway ponds recycling waste/exhaust streams to produce bulk chemicals for fuel, feed and industrial material.
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| 6. |
- Sánchez I Nogué, Violeta, et al.
(författare)
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Isolation and characterization of a resident tolerant Saccharomyces cerevisiae strain from a spent sulfite liquor fermentation plant
- 2012
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Ingår i: AMB Express. - : Springer Science and Business Media LLC. - 2191-0855. ; 2:1, s. 68-
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Tidskriftsartikel (refereegranskat)abstract
- Spent Sulfite Liquor (SSL) from wood pulping facilities is a sugar rich effluent that can be used as feedstock for ethanol production. However, depending on the pulping process conditions, the release of monosaccharides also generates a range of compounds that negatively affect microbial fermentation. In the present study, we investigated whether endogenous yeasts in SSL-based ethanol plant could represent a source of Saccharomyces cerevisiae strains with a naturally acquired tolerance towards this inhibitory environment. Two isolation processes were performed, before and after the re-inoculation of the plant with a commercial baker’s yeast strain. The isolates were clustered by DNA fingerprinting and a recurrent Saccharomyces cerevisiae strain, different from the inoculated commercial baker’s yeast strain, was isolated. The strain, named TMB3720, flocculated heavily and presented high furaldehyde reductase activity. During fermentation of undiluted SSL, TMB3720 displayed a 4-fold higher ethanol production rate and 1.8-fold higher ethanol yield as compared to the commercial baker’s yeast. Another non-Saccharomyces cerevisiae species, identified as the pentose utilizing Pichia galeiformis, was also recovered in the last tanks of the process where the hexose to pentose sugar ratio and the inhibitory pressure are expected to be the lowest.
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| 7. |
- Tomas-Pejo, Elia, 1980, et al.
(författare)
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EVALUATION OF EVOLVED AND BARCODED XYLOSE FERMENTING STRAINS FOR BIOETHANOL PRODUCTION FROM LIGNOCELLULOSE
- 2012
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Ingår i: Science and Technology Day 2012, Chalmers University of Technology, 27th March 2012.
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Lignocellulosic raw materials for bioethanol production are today the basis for many ethanol production sites around the world. However, the utilization of engineered yeast strains for second generation ethanol production at large-scale can still be improved. Yeasts mainly use the sugars present in the lignocellulosic biomass but, toxic compounds derived from cellulose, hemicellulose and lignin degradation during pretreatment are also found in the media and inhibit yeast growth. Furthermore, wild type Saccharomyces cerevisiae is not able to ferment xylose which could constitute up to 40% of the lignocellulose material. Hence the recombinant yeast strains must be robust and ferment xylose to ethanol with high yields in the presence of inhibitors.In this study, different evolved xylose fermenting Saccharomyces cerevisiae strains have been compared in ethanol production processes from lignocellulosic hydrolysates. The differences between using single cell transformants and mixed populations will be evaluated in terms of ethanol production in large scale bioreactors.Furthermore, we have established a method to barcode the evolved yeast strains in order to be able to verify their origin. It is of outmost importance that after barcoding the original characteristics of a yeast strain are maintained. Those requirements can only be fulfilled by using a dominant selection principle. We have obtained a few hundred transformants that were shown to contain the new unique barcode DNA sequence via DNA isolation and DNA sequencing. The transformed strains must be able to grow on the lignocellulosic material and consume xylose at the same rate as before the transformation which also was tested in this study.
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| 8. |
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| 9. |
- Martinez Avila, Hector, 1985, et al.
(författare)
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Biocompatibility evaluation of densified bacterial nanocellulose hydrogel as an implant material for auricular cartilage regeneration
- 2014
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Ingår i: Applied Microbiology and Biotechnology. - : Springer Science and Business Media LLC. - 1432-0614 .- 0175-7598. ; 98:17, s. 7423-7435
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Tidskriftsartikel (refereegranskat)abstract
- Bacterial nanocellulose (BNC), synthesized by the bacterium Gluconacetobacter xylinus, is composed of highly hydrated fibrils (99 % water) with high mechanical strength. These exceptional material properties make BNC a novel biomaterial for many potential medical and tissue engineering applications. Recently, BNC with cellulose content of 15 % has been proposed as an implant material for auricular cartilage replacement, since it matches the mechanical requirements of human auricular cartilage. This study investigates the biocompatibility of BNC with increased cellulose content (17 %) to evaluate its response in vitro and in vivo. Cylindrical BNC structures (48 Au 20 mm) were produced, purified in a built-in house perfusion system, and compressed to increase the cellulose content in BNC hydrogels. The reduction of endotoxicity of the material was quantified by bacterial endotoxin analysis throughout the purification process. Afterward, the biocompatibility of the purified BNC hydrogels with cellulose content of 17 % was assessed in vitro and in vivo, according to standards set forth in ISO 10993. The endotoxin content in non-purified BNC (2,390 endotoxin units (EU)/ml) was reduced to 0.10 EU/ml after the purification process, level well below the endotoxin threshold set for medical devices. Furthermore, the biocompatibility tests demonstrated that densified BNC hydrogels are non-cytotoxic and cause a minimal foreign body response. In support with our previous findings, this study concludes that BNC with increased cellulose content of 17 % is a promising non-resorbable biomaterial for auricular cartilage tissue engineering, due to its similarity with auricular cartilage in terms of mechanical strength and host tissue response.
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| 10. |
- Svensson, Sven-Erik, et al.
(författare)
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Ethanol production from industrial hemp : Effect of combined dilute acid/steam pretreatment and economic aspects
- 2014
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Ingår i: Bioresource Technology. - : Elsevier BV. - 0960-8524 .- 1873-2976. ; 163, s. 236-243
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Tidskriftsartikel (refereegranskat)abstract
- In the present study, combined steam (140-180 degrees C) and dilute-acid pre-hydrolysis (0.0-2.0%) were applied to industrial hemp (Cannabis sativa L), as pretreatment for lignocellulosic bioethanol production. The influence of the pretreatment conditions and cultivation type on the hydrolysis and ethanol yields was also evaluated. Pretreatment with 1% sulfuric acid at 180 degrees C resulted in the highest glucose yield (73-74%) and ethanol yield of 75-79% (0.38-0.40 g-ethanol/g-glucose). Taking into account the costs of biomass processing, from field to ethanol facility storage, the field-dried hemp pretreated at the optimal conditions showed positive economic results. The type of hemp cultivation (organic or conventional) did not influence significantly the effectiveness of the pretreatment as well as subsequent enzymatic hydrolysis and ethanol fermentation. (C) 2014 Elsevier Ltd. All rights reserved.
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