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  • 1.
    Abrahamsson, Louise
    Linköping University, Department of Thematic Studies, Tema Environmental Change.
    Improving methane production using hydrodynamic cavitation as pre-treatment2016Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    To develop anaerobic digestion (AD), innovative solutions to increase methane yields in existing AD processes are needed. In particular, the adoption of low energy pre-treatments to enhance biomass biodegradability is needed to provide efficient digestion processes increasing profitability. To obtain these features, hydrodynamic cavitation has been evaluated as an innovative solutions for AD of waste activated sludge (WAS), food waste (FW), macro algae and grass, in comparison with steam explosion (high energy pre-treatment). The effect of these two pre-treatments on the substrates, e.g. particle size distribution, soluble chemical oxygen demand (sCOD), biochemical methane potential (BMP) and biodegradability rate, have been evaluated. After two minutes of hydrodynamic cavitation (8 bar), the mean fine particle size decreased from 489- 1344 nm to 277- 381 nm (≤77% reduction) depending of the biomasses. Similar impacts were observed after ten minutes of steam explosion (210 °C, 30 bar) with a reduction in particle size between 40% and 70% for all the substrates treated.  In terms of BMP value, hydrodynamic cavitation caused significant increment only within the A. nodosum showing a post treatment increment of 44% compared to the untreated value, while similar values were obtained before and after treatment within the other tested substrates. In contrast, steam explosion allowed an increment for all treated samples, A. nodosum (+86%), grass (14%) and S. latissima (4%). However, greater impacts where observed with hydrodynamic cavitation than steam explosion when comparing the kinetic constant K. Overall, hydrodynamic cavitation appeared an efficient pre-treatment for AD capable to compete with the traditional steam explosion in terms om kinetics and providing a more efficient energy balance (+14%) as well as methane yield for A. nodosum.

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  • 2.
    Agarwal, Parminder
    et al.
    Michigan State University.
    Berglund, Kris
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Effect of polymeric additives on calcium carbonate crystallization as monitored by nephelometry2004In: Crystal Growth & Design, ISSN 1528-7483, E-ISSN 1528-7505, Vol. 4, no 3, p. 479-483Article in journal (Refereed)
    Abstract [en]

    The effect of polymaleimide polymers on calcium carbonate crystallization was studied using nephelometry. Induction time and percent growth inhibition were determined for polymeric additives from the nephelometric data. The polymaleimide synthesized by KOH-initiated polymerization exhibited the greatest growth inhibition and longest nucleation time among the polymers investigated. Raman spectroscopy was used to determine the calcium carbonate polymorph formed in the presence of these polymeric additives.

  • 3.
    Agarwal, Parminder
    et al.
    Michigan State University.
    Berglund, Kris
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    In situ monitoring of calcium carbonate polymorphs during batch crystallization in the presence of polymeric additives using Raman spectroscopy2003In: Crystal Growth & Design, ISSN 1528-7483, E-ISSN 1528-7505, Vol. 3, no 6, p. 941-946Article in journal (Refereed)
    Abstract [en]

    Polycarboxylic acids are well-known to affect calcium carbonate crystallization. Agarwal et al. (Ind. Eng. Chem. Res. 2003, in press) reported previously the synthesis of polymaleimide by a variety of techniques and initiators. In the present work, the effect of these polymers on calcium carbonate crystallization was studied by a variety of techniques. Crystallization experiments were carried out in a 1-L LABMAX automated batch reactor, and the concentration of calcium in solution was determined in real time. Raman spectroscopy was used to determine the relative amount of various calcium carbonate polymorphs as the crystallization occurred. However, Raman spectroscopy is a scattering technique, which may make it surface selective, and therefore results from solids may not be representative of bulk of sample. X-ray diffraction (XRD) was used to compare the results obtained by Raman spectroscopy. Peak intensity ratios were used for both Raman spectroscopy and XRD for calibration and measurement purposes. The results obtained by these two techniques for final percent vaterite for calcium carbonate crystallization in the presence of polymeric additives were in agreement within 2%. Therefore, use of Raman spectroscopy for in situ measurement of polymorph composition during calcium carbonate crystallization appears accurate. Scanning electron microscopy (SEM) data were useful in understanding the crystal morphology and to determine crystal size.

  • 4.
    Agarwal, Parminder
    et al.
    Michigan State University.
    Yu, Qiuyue
    Michigan State University.
    Harant, Adam
    Michigan State University.
    Berglund, Kris
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Synthesis and characterization of polymaleimide2003In: Industrial & Engineering Chemistry Research, ISSN 0888-5885, E-ISSN 1520-5045, Vol. 42, no 13, p. 2881-2884Article in journal (Refereed)
    Abstract [en]

    Simplified syntheses of polymaleimide employing anionic polymerization (from the melt and from solution) and metal compound-alcohol initiators such as PbO, SnO, tin bis(2-ethyl hexanoate) in the presence of tert-butyl benzyl alcohol are presented. The resulting polymers contain a combination of C-N- and C-C-connected monomers. Preliminary structures of the polymers were determined using NMR spectroscopy. The ratio of C-N- and C-C-connected monomers was determined, and the percentage of C-N-connected monomer units was found to vary from 40 to 80%, with the higher percentage resulting from anionic polymerization. The molecular weights of the polymers, as determined by gel permeation chromatography (GPC) with aqueous mobile phase and sodium polyacrylates standards, ranged between 1100 and 4200 for anionic polymerization and were about 11 500 for metal oxide-alcohol initiated polymerization. Solution-phase properties of the polymaleimides were evaluated by calcium chelation and precipitation inhibition studies. On the basis of the measured properties of these polymers, they are proposed as biodegradable, low-impact detergent additives to substitute currently used compounds.

  • 5.
    Agnihotri, Swarnima
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Yin, D M
    Institute of Urban and Rural Mining, Changzhou University, Changzhou, China.
    Mahboubi, Amir
    University of Borås, Faculty of Textiles, Engineering and Business.
    Sapmaz, Tugba
    University of Borås, Faculty of Textiles, Engineering and Business.
    Varjani, S
    Gujarat Pollution Control Board, Gandhinagar, India.
    Qiao, W
    Institute of Urban and Rural Mining, Changzhou University, Changzhou, China.
    Koseoglu-Imer, D Y
    Department of Environmental Engineering, Istanbul Technical University, Maslak, Istanbul, Turkey.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    A Glimpse of the World of Volatile Fatty Acids Production and Application: A review2022In: Bioengineered, ISSN 2165-5979, E-ISSN 2165-5987, Vol. 13, no 1, p. 1249-1275Article, review/survey (Refereed)
    Abstract [en]

    Sustainable provision of chemicals and materials is undoubtedly a defining factor in guaranteeing economic, environmental, and social stability of future societies. Among the most sought-after chemical building blocks are volatile fatty acids (VFAs). VFAs such as acetic, propionic, and butyric acids have numerous industrial applications supporting from food and pharmaceuticals industries to wastewater treatment. The fact that VFAs can be produced synthetically from petrochemical derivatives and also through biological routes, for example, anaerobic digestion of organic mixed waste highlights their provision flexibility and sustainability. In this regard, this review presents a detailed overview of the applications associated with petrochemically and biologically generated VFAs, individually or in mixture, in industrial and laboratory scale, conventional and novel applications.

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  • 6.
    Ahlén, Gustaf
    et al.
    Recopharma AB, Stockholm, Sweden.
    Strindelius, Lena
    Recopharma AB, Stockholm, Sweden.
    Johansson, Tomas
    Recopharma AB, Stockholm, Sweden.
    Nilsson, Anki
    Recopharma AB, Stockholm, Sweden.
    Chatzissavidou, Nathalie
    Recopharma AB, Stockholm, Sweden.
    Sjöblom, Magnus
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Holgersson, Jan
    Clinical Chemistry and Transfusion Medicine, Sahlgrenska Academy, Gothenburg, Sweden.
    Mannosylated mucin-type immunoglobulin fusion proteins enhance antigen-specific antibody and T lymphocyte responses2012In: PLOS ONE, E-ISSN 1932-6203, Vol. 7, no 10, article id e46959Article in journal (Refereed)
    Abstract [en]

    Targeting antigens to antigen-presenting cells (APC) improve their immunogenicity and capacity to induce Th1 responses and cytotoxic T lymphocytes (CTL). We have generated a mucin-type immunoglobulin fusion protein (PSGL-1/mIgG2b), which upon expression in the yeast Pichia pastoris became multivalently substituted with O-linked oligomannose structures and bound the macrophage mannose receptor (MMR) and dendritic cell-specific intercellular adhesion molecule-3 grabbing non-integrin (DC-SIGN) with high affinity in vitro. Here, its effects on the humoral and cellular anti-ovalbumin (OVA) responses in C57BL/6 mice are presented.

    OVA antibody class and subclass responses were determined by ELISA, the generation of anti-OVA CTLs was assessed in 51Cr release assays using in vitro-stimulated immune spleen cells from the different groups of mice as effector cells and OVA peptide-fed RMA-S cells as targets, and evaluation of the type of Th cell response was done by IFN-γ, IL-2, IL-4 and IL-5 ELISpot assays.

    Immunizations with the OVA − mannosylated PSGL-1/mIgG2b conjugate, especially when combined with the AbISCO®-100 adjuvant, lead to faster, stronger and broader (with regard to IgG subclass) OVA IgG responses, a stronger OVA-specific CTL response and stronger Th1 and Th2 responses than if OVA was used alone or together with AbISCO®-100. Also non-covalent mixing of mannosylated PSGL-1/mIgG2b, OVA and AbISCO®-100 lead to relatively stronger humoral and cellular responses. The O-glycan oligomannoses were necessary because PSGL-1/mIgG2b with mono- and disialyl core 1 structures did not have this effect.

    Mannosylated mucin-type fusion proteins can be used as versatile APC-targeting molecules for vaccines and as such enhance both humoral and cellular immune responses.

  • 7.
    Akinbomi, J G
    et al.
    Department of Chemical Engineering, Faculty of Engineering, Lagos State University, Lagos, 100268, Nigeria.
    Patinvoh, R J
    Department of Chemical Engineering, Faculty of Engineering, Lagos State University, Lagos, 100268, Nigeria.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Current challenges of high-solid anaerobic digestion and possible measures for its effective applications: a review2022In: Biotechnology for Biofuels and Bioproducts, E-ISSN 2731-3654, Vol. 15, no 1Article, review/survey (Refereed)
    Abstract [en]

    The attention that high solids anaerobic digestion process (HS-AD) has received over the years, as a waste management and energy recovery process when compared to low solids anaerobic digestion process, can be attributed to its associated benefits including water conservation and smaller digester foot print. However, high solid content of the feedstock involved in the digestion process poses a barrier to the process stability and performance if it is not well managed. In this review, various limitations to effective performance of the HS-AD process, as well as, the possible measures highlighted in various research studies were garnered to serve as a guide for effective industrial application of this technology. A proposed design concept for overcoming substrate and product inhibition thereby improving methane yield and process stability was recommended for optimum performance of the HS-AD process.

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  • 8.
    Alinaghi, Masoumeh
    et al.
    Umeå University. Functional Microbiology, Institute of Microbiology, Department of Pathobiology, University of Veterinary Medicine, Vienna, Austria.
    Nilsson, David
    Umeå University, Faculty of Science and Technology, Department of Chemistry.
    Singh, Nikita
    Umeå University, Faculty of Science and Technology, Department of Chemistry.
    Höjer, Annika
    Norrmejerier, Umeå, Sweden.
    Saedén, Karin Hallin
    Norrmejerier, Umeå, Sweden.
    Trygg, Johan
    Umeå University, Faculty of Science and Technology, Department of Chemistry. Sartorius Corporate Research, Sartorius, Sartorius Stedim Data Analytics, Umeå, Sweden.
    Near-infrared hyperspectral image analysis for monitoring the cheese-ripening process2023In: Journal of Dairy Science, ISSN 0022-0302, E-ISSN 1525-3198, Vol. 106, no 11, p. 7407-7418Article in journal (Refereed)
    Abstract [en]

    Ripening is the most crucial process step in cheese manufacturing and constitutes multiple biochemical alterations that describe the final cheese quality and its perceived sensory attributes. The assessment of the cheese-ripening process is challenging and requires the effective analysis of a multitude of biochemical changes occurring during the process. This study monitored the biochemical and sensory attribute changes of paraffin wax-covered long-ripening hard cheeses (n = 79) during ripening by collecting samples at different stages of ripening. Near-infrared hyperspectral (NIR-HS) imaging, together with free amino acid, chemical composition, and sensory attributes, was studied to monitor the biochemical changes during the ripening process. Orthogonal projection-based multivariate calibration methods were used to characterize ripening-related and orthogonal components as well as the distribution map of chemical components. The results approve the NIR-HS imaging as a rapid tool for monitoring cheese maturity during ripening. Moreover, the pixelwise evaluation of images shows the homogeneity of cheese maturation at different stages of ripening. Among the chemical compositions, fat content and moisture are the most important variables correlating to NIR-HS images during the ripening process.

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  • 9.
    Alinejad, M.
    et al.
    Department of Forestry, Michigan State University, East Lansing, United States.
    Henry, C.
    Department of Forestry, Michigan State University, East Lansing, United States.
    Nikafshar, S.
    Department of Forestry, Michigan State University, East Lansing, United States.
    Gondaliya, A.
    Chemical Engineering and Materials Science, Michigan State University, East Lansing, United States.
    Bagheri, B.
    Chemical Engineering and Materials Science, Michigan State University, East Lansing, United States.
    Chen, N.
    Eastern Regional Research Center, USDA-ARS, Wyndmoor, United States.
    Singh, S.K.
    Chemical and Biological Engineering, Montana State University, Bozeman, United States.
    Hodge, David
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Nejad, M.
    Department of Forestry, Michigan State University, East Lansing, United States. Chemical Engineering and Materials Science, Michigan State University, East Lansing, United States..
    Lignin-based polyurethanes: Opportunities for bio-based foams, elastomers, coatings and adhesives2019In: Polymers, E-ISSN 2073-4360, Vol. 11, no 7, article id 1202Article in journal (Refereed)
    Abstract [en]

    Polyurethane chemistry can yield diverse sets of polymeric materials exhibiting a widerange of properties for various applications and market segments. Utilizing lignin as a polyol presentsan opportunity to incorporate a currently underutilized renewable aromatic polymer into theseproducts. In this work, we will review the current state of technology for utilizing lignin as a polyolreplacement in different polyurethane products. This will include a discussion of lignin structure,diversity, and modification during chemical pulping and cellulosic biofuels processes, approachesfor lignin extraction, recovery, fractionation, and modification/functionalization. We will discussthe potential of incorporation of lignins into polyurethane products that include rigid and flexiblefoams, adhesives, coatings, and elastomers. Finally, we will discuss challenges in incorporating ligninin polyurethane formulations, potential solutions and approaches that have been taken to resolvethose issues.

  • 10.
    Amorim, Lúcia F.A.
    et al.
    FibEnTech Research Unit, Faculty of Engineering, University of Beira Interior, Covilhã, Portugal.
    Li, Lengwan
    KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Centres, Wallenberg Wood Science Center. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Fibre- and Polymer Technology, Biocomposites.
    Gomes, Ana P.
    FibEnTech Research Unit, Faculty of Engineering, University of Beira Interior, Covilhã, Portugal.
    Fangueiro, Raul
    Centre for Textile Science and Technology (2C2T), University of Minho, Guimarães, Portugal.
    Gouveia, Isabel C.
    FibEnTech Research Unit, Faculty of Engineering, University of Beira Interior, Covilhã, Portugal.
    Sustainable bacterial cellulose production by low cost feedstock: evaluation of apple and tea by-products as alternative sources of nutrients2023In: Cellulose, ISSN 0969-0239, E-ISSN 1572-882X, Vol. 30, no 9, p. 5589-5606Article in journal (Refereed)
    Abstract [en]

    The high applicability of Bacterial Cellulose (BC) is often challenging due to its high production costs, which ultimately prevents its widespread use. Therefore, the present study aimed to investigate BC production using alternative feedstock to replace high-cost synthetic carbon and nitrogen sources and to evaluate the physical and structural properties of the produced BC membranes. BC was produced through a microbial consortium from kombucha, and the formulated alternative media sustained promising BC production, especially the association of apple wastes (at 10% (W/V)) with tea mixture, with a yield similar to BC produced on Hestrin–Schramm (HS) control media. Moreover, the BC samples produced in this alternative media also exhibited comparable properties to BC from HS media, with similar water-holding capacity and retention ability, thermal stability, mechanical behavior, and a crystallinity index of 87.61% and 88.08%, respectively. Thus, our findings substantiated that expensive substrates, such as glucose, peptone, and yeast extract, could be successfully replaced by apple wastes, black and green tea, for BC production while maintaining its remarkable physical and structural properties. Furthermore, besides the low-cost advantage, the bioconversion of apple waste also reduces the environmental burden caused by its disposal in landfills.

  • 11.
    Anasontzis, George E
    et al.
    Industrial Biotechnology, Chemical and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden; Wallenberg Wood Science Center, Chalmers University of Technology, Gothenburg, Sweden.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Challenges in ethanol production with Fusarium oxysporum through consolidated bioprocessing2014In: Bioengineered, ISSN 2165-5979, E-ISSN 2165-5987, Vol. 5, no 6, p. 393-395Article in journal (Other (popular science, discussion, etc.))
    Abstract [en]

    Fusarium oxysporum has been reported as being able to both produce the enzymes necessary to degrade lignocellulosic biomass to sugars and also ferment the monosaccharides to ethanol under anaerobic or microaerobic conditions. However, in order to become an economically feasible alternative to other ethanol-producing microorganisms, a better understanding of its physiology, metabolic pathways, and bottlenecks is required, together with an improvement in its efficiency and robustness. In this report, we describe the challenges for the future and give additional justification for our recent publication.

  • 12.
    Anasontzis, George E.
    et al.
    National and Kapodistrian University of Athens, Chalmers University of Technology, Department of Chemical and Biological Engineering, Microbial Biotechnology Unit, Sector of Botany, Department of Biology, National and Kapodistrian University of Athens, Zografou.
    Kourtoglou, Elisavet
    National Technical University of Athens, BIOtechMASS Unit, Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Villas-Boâs, Silas G
    Centre for Microbial Innovation, School of Biological Sciences, The University of Auckland, Technical University of Denmark.
    Hatzinikolaou, Dimitris G.
    Department of Chemical Engineering, National Technical University of Athens.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Metabolic Engineering of Fusarium oxysporum to Improve Its Ethanol-Producing Capability2016In: Frontiers in Microbiology, E-ISSN 1664-302X, Vol. 7, article id 632Article in journal (Refereed)
    Abstract [en]

    Fusarium oxysporum is one of the few filamentous fungi capable of fermenting ethanol directly from plant cell wall biomass. It has the enzymatic toolbox necessary to break down biomass to its monosaccharides and, under anaerobic and microaerobic conditions, ferments them to ethanol. Although these traits could enable its use in consolidated processes and thus bypass some of the bottlenecks encountered in ethanol production from lignocellulosic material when Saccharomyces cerevisiae is used-namely its inability to degrade lignocellulose and to consume pentoses-two major disadvantages of F. oxysporum compared to the yeast-its low growth rate and low ethanol productivity-hinder the further development of this process. We had previously identified phosphoglucomutase and transaldolase, two major enzymes of glucose catabolism and the pentose phosphate pathway, as possible bottlenecks in the metabolism of the fungus and we had reported the effect of their constitutive production on the growth characteristics of the fungus. In this study, we investigated the effect of their constitutive production on ethanol productivity under anaerobic conditions. We report an increase in ethanol yield and a concomitant decrease in acetic acid production. Metabolomics analysis revealed that the genetic modifications applied did not simply accelerate the metabolic rate of the microorganism; they also affected the relative concentrations of the various metabolites suggesting an increased channeling toward the chorismate pathway, an activation of the γ-aminobutyric acid shunt, and an excess in NADPH regeneration

  • 13.
    Anasontzis, George
    et al.
    Chalmers University of Technology, Department of Chemical and Biological Engineering.
    Kourtoglou, Elisavet
    BIOtechMASS Unit, Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Mamma, Diomi
    BIOtechMASS Unit, Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Villas-Boâs, Silas G
    Centre for Microbial Innovation, School of Biological Sciences, The University of Auckland.
    Hatzinikolaou, Dimitris
    Microbial Biotechnology Unit, Sector of Botany, Department of Biology, National and Kapodistrian University of Athens.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Constitutive homologous expression of phosphoglucomutase and transaldolase increases the metabolic flux of Fusarium oxysporum2014In: Microbial Cell Factories, E-ISSN 1475-2859, Vol. 13, article id 43Article in journal (Refereed)
    Abstract [en]

    BACKGROUND: Fusarium oxysporum is among the few filamentous fungi that have been reported of being able to directly ferment biomass to ethanol in a consolidated bioprocess. Understanding its metabolic pathways and their limitations can provide some insights on the genetic modifications required to enhance its growth and subsequent fermentation capability. In this study, we investigated the hypothesis reported previously that phosphoglucomutase and transaldolase are metabolic bottlenecks in the glycolysis and pentose phosphate pathway of the F. oxysporum metabolism.RESULTS: Both enzymes were homologously overexpressed in F. oxysporum F3 using the gpdA promoter of Aspergillus nidulans for constitutive expression. Transformants were screened for their phosphoglucomutase and transaldolase genes expression levels with northern blot. The selected transformant exhibited high mRNA levels for both genes, as well as higher specific activities of the corresponding enzymes, compared to the wild type. It also displayed more than 20 and 15% higher specific growth rate upon aerobic growth on glucose and xylose, respectively, as carbon sources and 30% higher xylose to biomass yield. The determination of the relative intracellular amino and non-amino organic acid concentrations at the end of growth revealed higher abundance of most determined metabolites between 1.5- and 3-times in the recombinant strain compared to the wild type. Lower abundance of the determined metabolites of the Krebs cycle and an 68-fold more glutamate were observed at the end of the cultivation, when xylose was used as carbon source.CONCLUSIONS: Homologous overexpression of phosphoglucomutase and transaldolase in F. oxysporum was shown to enhance the growth characteristics of the strain in both xylose and glucose in aerobic conditions. The intracellular metabolites profile indicated how the changes in the metabolome could have resulted in the observed growth characteristics.

  • 14.
    Anbalagan, Anbarasan
    Mälardalen University, School of Business, Society and Engineering, Future Energy Center.
    A passage to wastewater nutrient recovery units: Microalgal-Bacterial bioreactors2018Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    In recent years, the microalgal–bacterial process has been considered to be a very attractive engineering solution for wastewater treatment. However, it has not been widely studied in the context of conventional wastewater treatment design under Swedish conditions. The technology holds several advantages: as a CO2 sink, ability to withstand cold conditions, ability to grow under low light, fast settling without chemical precipitation, and reducing the loss of valuable nutrients (CO2, N2, N2O, PO4). The process also provides the option to be operated either as mainstream (treatment of municipal wastewater) or side stream (treatment of centrate from anaerobic digesters) to reduce the nutrient load of the wastewater. Furthermore, the application is not only limited to wastewater treatment; the biomass can be used to synthesise platform chemicals or biofuels and can be followed by recovery of ammonium and phosphate for use in agriculture.

    In the present study, the feasibility of applying the process in Swedish temperature and light conditions was investigated by implementing microalgae within the activated sludge process. In this context, the supporting operational and performance indicators (hydraulic retention time (HRT), sludge retention time (SRT) and nutrients removal) were evaluated to support naturally occurring consortia in photo-sequencing and continuous bioreactor configuration. Furthermore, CO2 uptake and light spectrum-mediated nutrient removal were investigated to reduce the impact on climate and the technical challenges associated with this type of system.

    The results identified effective retention times of 6 and 4 days (HRT = SRT) under limited lighting to reduce the electrical consumption. From the perspective of nitrogen removal, the process demands effective CO2 input either in the mainstream or side stream treatment. The incorporation of a vertical absorption column demonstrated effective CO2 mass transfer to support efficient nitrogen and phosphorus removal as a side stream treatment. However, the investigation of a continuous single-stage process as the mainstream showed a requirement for a lower SRT in comparison to semi-continuous operation due to faster settlability, regardless of inorganic carbon. Furthermore, the process showed an effective reduction of influent phosphorus and organic compounds (i.e. COD/TOC) load in the wastewater as a result of photosynthetic aeration. Most importantly, the operation was stable at the temperature equivalent of wastewater (12 and 13 ˚C), under different lighting (white, and red-blue wavelengths) and retention times (6 and 1.5 d HRT) with complete nitrification. Additionally, the biomass production was stable with faster settling properties without any physiochemical separation.

    The outcomes of this thesis on microalgal–bacterial nutrient removal demonstrates that (1) photosynthesis-based aeration at existing wastewater conditions under photo-sequential and continuous photobioreactor setup, (2) flocs with rapid settling characteristics at all studied retention times, (3) the possibility of increasing carbon supplementation to achieve higher carbon to nitrogen balance in the photobioreactor, and (4) most importantly, nitrification-based microalgal biomass uptake occurred at all spectral distributions, lower photosynthetic active radiation and existing wastewater conditions.

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  • 15.
    Anderson, Mattias
    et al.
    KTH, School of Biotechnology (BIO), Industrial Biotechnology.
    Afewerki, Samson
    Berglund, Per
    KTH, School of Biotechnology (BIO), Industrial Biotechnology.
    Cõrdova, Armando
    Total Synthesis of Capsaicin Analogues from Lignin-Derived Compounds by Combined Heterogeneous Metal, Organocatalytic and Enzymatic Cascades in One Pot2014In: Advanced Synthesis and Catalysis, ISSN 1615-4150, E-ISSN 1615-4169, Vol. 356, no 9, p. 2113-2118Article in journal (Refereed)
    Abstract [en]

    The total synthesis of capsaicin analogues was performed in one pot, starting from compounds that can be derived from lignin. Heterogeneous palladium nanoparticles were used to oxidise alcohols to aldehydes, which were further converted to amines by an enzyme cascade system, including an amine transaminase. It was shown that the palladium catalyst and the enzyme cascade system could be successfully combined in the same pot for conversion of alcohols to amines without any purification of intermediates. The intermediate vanillyl-amine, prepared with the enzyme cascade system, could be further converted to capsaicin analogues without any purification using either fatty acids and a lipase, or Schotten-Baumann conditions, in the same pot. An aldol compound (a simple lignin model) could also be used as starting material for the synthesis of capsaicin analogues. Using l-alanine as organocatalyst, vanillin could be obtained by a retro-aldol reaction. This could be combined with the enzyme cascade system to convert the aldol compound to vanillylamine in a one-step one-pot reaction.

  • 16.
    Andersson, Christian
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Biobased production of succinic acid by Escherichia coli fermentation2009Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The prospects of peak oil, climate change and the dependency of fossil carbon have urged research and development of production methods for the manufacture of fuels and chemicals from renewable resources (biomass). The present thesis illustrates different aspects of biobased succinic acid production by a metabolically engineered E. coli strain. The main areas of the thesis are sugar utilisation and feedstock flexibility, and fermentation inhibition, both due to toxic compound derived from the raw material and the fermentation products themselves.The first part of this thesis aimed to investigate the fermentation characteristics of AFP184 in a medium consisting of corn steep liquor, inorganic salts and different sugar sources without supplementation with high-cost nutrients such as yeast extract and peptone. The effects of different sugars, sucrose, glucose, fructose, xylose, equal mixtures of glucose-fructose and glucose-xylose, on succinic acid production kinetics and yields in an industrially relevant medium were investigated. AFP184 was able to utilise all sugars and sugar combinations except sucrose for biomass generation and succinate production. Using glucose resulted in the highest yield, 0.83 (g succinic acid per g sugar consumed anaerobically). Using a high initial sugar concentration resulted in volumetric productivities of almost 3 g L-1 h-1, which is above estimated values for economically feasible production. However, succinic acid production ceased at final concentrations greater than 40 g L-1. To further increase succinic acid concentrations, fermentations using NH4OH, NaOH, KOH, K2CO3, and Na2CO3 as neutralising agents were performed and compared. It was shown that substantial improvements could be made by using alkali bases to neutralise the fermentations. The highest concentrations and productivities were achieved when Na2CO3 was used, 77 g L-1 and 3 g L-1 h-1 respectively. A gradual decrease in succinate productivity was observed during the fermentations, which was shown to be due to succinate accumulation in the broth and not as a result of the addition of neutralising agent or the subsequent increase in osmolarity.To maintain high succinate productivity by keeping a low extracellular succinic acid concentration fermentations were interrupted and cells recovered and resuspended in fresh media. By removing the succinate it was possible to maintain high succinic acid productivity for a prolonged time. Cells subjected to high concentrations of succinate were also able to regain high productivity once transferred into a succinate-free medium.In the last part of the thesis succinic acid production from softwood dilute acid hydrolysates was demonstrated. This study involved establishing the degree of detoxification necessary for growth and fermentation using industrial hydrolysates. Detoxification by treatment with lime and/or activated carbon was investigated and the results show that it was possible to produce succinate from softwood hydrolysates in yields comparable to those for synthetic sugars.The work done in this thesis increases the understanding of succinic acid production with AFP184, illustrate its limitations, and suggests improvements in the current technology with the long term aim of increasing the economical feasibility of biochemical succinic acid production.

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  • 17. Andersson, Christian
    Succinic acid production using metabolically engineered Escherichia coli2007Licentiate thesis, comprehensive summary (Other academic)
    Abstract [en]

    The prospects of peak oil, climate change and the dependency of fossil carbon have urged research and development of production methods for the manufacture of fuels and chemicals from renewable resources (biomass). To date, the primary emphasis has been placed on the replacement of oil for transportation fuels. A highly significant subset of petroleum usage is the production of chemicals, which represents 10-15% of the petroleum usage. White biotechnology, also called industrial biotechnology, is a fast evolving technology with a large potential to have a substantial impact on the industrial production of fuels and chemicals from biomass. This work addresses the issue of chemical production by investigating the production of bio-based succinic acid, which can be used in a wide range of applications to replace petroleum based chemicals. Succinic acid can be produced by fermentation of sugar by a number of organisms; one is Escherichia coli (E. coli). It is known that E. coli under anaerobic conditions produces a mixture of organic acids. In order to obtain a cost-effective production it is necessary to metabolically engineer the organism to produce succinic acid in greater yield than the other acids. In the current work, E. coli mutant AFP184 was used. AFP184 originates from a near wild type strain, the C600 (ATCC 23724), which can ferment both five and six carbon sugars and has mutations in the glucose specific phosphotransferase system (ptsG), the pyruvate formate lyase system (pfl) and in the fermentative lactate dehydrogenase system (ldh). The previous studies using different organisms have all used cultivation mediums supplemented to some degree with different nutrients like biotin, thiamine and yeast extract. In order to apply the technology to large scale, production must be cost-effective and it is important to minimise the use of additional supplements. The first part of this work aimed to investigate the fermentation characteristics of AFP184 in a medium consisting of corn steep liquor, inorganic salts and different sugar sources without supplementation of other additional nutrients. It addresses questions regarding the effect of different sugars on succinic acid kinetics and yields in an industrially relevant medium. In order to gain a sustainable production of succinic acid from biomass feedstocks (sugar from biomass) it is important to investigate how well the organism can utilise different sugars in the biomass. The sugars studied were sucrose, glucose, fructose, xylose and equal mixtures of glucose-fructose and glucose-xylose at a total initial sugar concentration of 100 g L-1. AFP184 was able to utilise all sugars and sugar combinations except sucrose for biomass generation and succinate production. Using glucose resulted in the highest yield, 0.83 (g succinic acid per g sugar consumed anaerobically). Fructose resulted in a yield of 0.66 and xylose of 0.5. Using a high initial sugar concentration made it possible to obtain volumetric productivities of almost 3 g L-1h-1, which is above estimated values for feasible economic production. Succinic acid production ceased at final concentrations greater than 40 g L-1. In order to further increase succinic acid concentrations, this inhibitory effect was studied in the second part of the present work. The inhibitory effects can be two-fold including pH-based inhibition and an anion specific effect on metabolism. It has been reported that high concentrations of ammonia inhibit E. coli growth and damage cell membranes. In order to limit toxic and inhibitory effects different neutralising agents were tested. First the use of NH4OH was optimised with respect to fermentation pH and it was found that the best results were obtained at pH 6.5-6.7. Optimal pH was then used with NaOH, KOH, and Na2CO3 as neutralising agents and it was shown that NaOH, KOH, and Na2CO3 neutralised fermentations could reach succinic acid concentrations of 69 and 61 and 78 g L-1 respectively without any significant decrease in succinic acid productivity. It was observed that cells lost viability during the cause anaerobic phase. It resulted in decreasing succinic acid productivities. It is believed that the viability decrease is a combined effect of organic acids concentration and the osmolarity of the medium. The work done in this thesis is aimed towards increasing the economical feasibility of a biochemical succinic acid production.

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  • 18. Andersson, Christian
    et al.
    Helmerius, Jonas
    Berglund, Kris
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Rova, Ulrika
    Effects of neutralising agent, organic acids, and osmolarity on succinic acid production by Escherichia coli AFP1842008Conference paper (Other academic)
    Abstract [en]

    Using a low-cost medium Escherichia coli AFP184 has previously been reported to produce succinic acid with volumetric productivities close to 3 g L-1 h-1. At a total organic acid concentration of 30 g L-1 the productivity decreased drastically resulting in final succinate concentrations of 40 g L-1. The economical viability of biochemical succinic acid production would benefit from higher final succinic acid concentrations and volumetric productivities maintained at >2.5 g L-1 h-1 for an extended period of time. In the present work the effects of osmolarity and neutralising agent (NH4OH, KOH, NaOH, K2CO3, and Na2CO3) on succinic acid production by AFP184 were investigated. Highest concentration of succinic acid was obtained with Na2CO3, 75 g L-1. It was also found that the osmolarity resulting from succinate production and subsequent base addition, only marginally affected the productivity per viable cell. Organic acid inhibition due to the produced succinic acid on the other hand significantly reduced succinic acid productivity per viable cell. When using NH4OH productivity completely ceased at approximately 40 g L-1. Volumetric productivities remained at 2.5 g L-1 h-1 for 5 to 10 hours longer when using K- or Na-bases than when using NH4OH. However, loss of cell viability occurred, and together with the acid inhibition decreased the volumetric productivities. In this study it was demonstrated that by altering the neutralising agent it was possible to increase the period of high volumetric productivity in the anaerobic phase and improve the final succinic acid concentration by almost 100 %

  • 19.
    Andersson, Christian
    et al.
    Luleå University of Technology.
    Helmerius, Jonas
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Hodge, David
    Luleå University of Technology.
    Berglund, Kris
    Luleå University of Technology.
    Rova, Ulrika
    Luleå University of Technology.
    Inhibition of succinic acid production in metabolically engineered Escherichia Coli by neutralizing agent, organic acids, and osmolarity2009In: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033, Vol. 25, no 1, p. 116-123Article in journal (Refereed)
    Abstract [en]

    The economical viability of biochemical succinic acid production is a result of many processing parameters including final succinic acid concentration, recovery of succinate, and the volumetric productivity. Maintaining volumetric productivities >2.5 g L-1 h(-1) is important if production of succinic acid from. renewable resources should be competitive. In this work, the effects of organic acids, osmolarity, and neutralizing agent (NH4OH, KOH, NaOH, K2CO3, and Na2CO3) on the fermentative succinic acid production by Escherichia coli AFP184 were investigated. The highest concentration of succinic acid, 77 g L-1. was obtained with Na2O3. In general, irrespective of the base used, succinic acid productivity per viable cell was significantly reduced as the concentration of the produced acid increased. Increased osmolarity resulting from base addition during succinate production only marginally affected the productivity per viable cell. Addition of the osmoprotectant glycine betaine to cultures resulted in an increased aerobic growth rate and anaerobic glucose consumption rate, but decreased succinic acid yield. When using NH4OH productivity completely ceased at a succinic acid concentration of similar to 40 g L-1. Volumetric productivities remained at 2.5 g L-1 h(-1) for tip to 10 h longer when K- or Na-bases where used instead of NH4OH. The decrease in cellular succinic acid productivity observed during the anaerobic phase was found to be due to increased organic acid concentrations rather than medium osmolarity.

  • 20. Andersson, Christian
    et al.
    Hodge, David
    Berglund, Kris
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Rova, Ulrika
    Effect of different carbon sources on the production of succinic acid using metabolically engineered Escherichia coli2007In: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033, Vol. 23, no 2, p. 381-388Article in journal (Refereed)
    Abstract [en]

    Succinic acid (SA) is an important platform molecule in the synthesis of a number of commodity and specialty chemicals. In the present work, dual-phase batch fermentations with the E. coli strain AFP184 were performed using a medium suited for large-scale industrial production of SA. The ability of the strain to ferment different sugars was investigated. The sugars studied were sucrose, glucose, fructose, xylose, and equal mixtures of glucose and fructose and glucose and xylose at a total initial sugar concentration of 100 g L-1. AFP184 was able to utilize all sugars and sugar combinations except sucrose for biomass generation and succinate production. For sucrose as a substrate no succinic acid was produced and none of the sucrose was metabolized. The succinic acid yield from glucose (0.83 g succinic acid per gram glucose consumed anaerobically) was higher than the yield from fructose (0.66 g g-1). When using xylose as a carbon source, a yield of 0.50 g g-1 was obtained. In the mixed-sugar fermentations no catabolite repression was detected. Mixtures of glucose and xylose resulted in higher yields (0.60 g g-1) than use of xylose alone. Fermenting glucose mixed with fructose gave a lower yield (0.58 g g-1) than fructose used as the sole carbon source. The reason is an increased pyruvate production. The pyruvate concentration decreased later in the fermentation. Final succinic acid concentrations were in the range of 25-40 g L-1. Acetic and pyruvic acid were the only other products detected and accumulated to concentrations of 2.7-6.7 and 0-2.7 g L-1. Production of succinic acid decreased when organic acid concentrations reached approximately 30 g L-1. This study demonstrates that E. coli strain AFP184 is able to produce succinic acid in a low cost medium from a variety of sugars with only small amounts of byproducts formed.

  • 21. Andersson, Christian
    et al.
    Lundberg, Angela
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Geosciences and Environmental Engineering.
    Distribution of snow cover accumulation: airborne ground penetrating radar surveys2002In: Proceedings of the XXII Nordic Hydrological Conference / [ed] Å. Killingtveit, 2002, p. 517-526Conference paper (Refereed)
  • 22.
    Andersson, Christian
    et al.
    Luleå University of Technology.
    Petrova, Ekaterina
    Luleå University of Technology.
    Berglund, Kris
    Luleå University of Technology. Departments of Forestry and Chemical Engineering, Michigan State University, Michigan, USA.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Maintaining high anaerobic succinic acid productivity by product removal2010In: Bioprocess and biosystems engineering (Print), ISSN 1615-7591, E-ISSN 1615-7605, Vol. 33, no 6, p. 711-718Article in journal (Refereed)
    Abstract [en]

    During dual-phase fermentations using Escherichia coli engineered for succinic acid production, the productivity and viable cell concentration decrease as the concentration of succinic acid increases. The effects of succinic acid on the fermentation kinetics, yield, and cell viability were investigated by resuspending cells in fresh media after selected fermentation times. The cellular succinic acid productivity could be restored, but cell viability continuously decreased throughout the fermentations by up to 80% and subsequently the volumetric productivity was reduced. Omitting complex nutrients in the resuspension media had no significant effect on cellular succinate productivity and yield, although the viable cell concentration and thus the volumetric productivity was reduced by approximately 20%. By resuspending the cells, the amount of succinate produced during a 100-h fermentation was increased by more than 60%. The results demonstrate that by product removal succinic acid productivity can be maintained at high levels for extended periods of time.

  • 23.
    Andersson, Christian
    et al.
    Luleå University of Technology.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Berglund, Kris
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Process for producing succinic acid from sucrose2005Patent (Other (popular science, discussion, etc.))
    Abstract [en]

    A process for hydrolyzing sucrose to glucose and fructose using succinic acid is described. The hydrolysate can be used to produce purified glucose and/or fructose or can be used as a carbon source for fermentations to produce various chemicals including succinic acid.

  • 24.
    Andersson, Klara
    KTH, School of Biotechnology (BIO).
    Development of a shake flask method suitable for effective screening of Escherichia coli expression constructs2011Independent thesis Advanced level (professional degree), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    Screening of expression constructs suitable for protein pharmaceuticals is often done in batch cultivations. But the production of the recombinant protein is made during fed-batch cultivations. The two types of cultivations are different and therefore may good expression constructs that grow poorly in batch cultivations but good in fed-batch cultivations be rejected. Therefore would it be desirable to develop a fed-batch method that can be used in shake flasks. Biosilta has developed a method where starch is broken down into glucose by an enzyme creating fed-batch conditions. This method has been tried out and analyzed during this project. It is shown that the cells grown under these conditions can be glucose limited. However, at a later stage of the cultivation the cells produce a large amount of acetate and pH is not stable. The system builds on a booster tablet which content is unknown. If the booster is not added to the cultivations the cells stop growing, this indicates that there is some other limitation than just glucose. It is also seen that the amount of protein that is produced during this fed-batch mimic cultivation is much lower than that is produced during normal batch cultivations. I would therefore not recommend EnBase as a screening method.

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  • 25.
    Andersson, Moa
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Edman, Linus
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Kredell, Lova
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Sandqvist, Tilda
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Eliasson, Johan
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Product sieving of monoclonal antibodies in cell culture processes: An investigation of product retention in perfusion cell cultures2024Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
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  • 26.
    Andraous, Johnny I.
    et al.
    Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing.
    Claus, Michael J
    Department of Agricultural Engineering, Michigan State University.
    Berglund, Kris
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Lindemann, Deirdre J.
    Department of Chemistry, Michigan State University.
    Effect of liquefaction enzymes on methanol concentration of distilled fruit spirits2004In: American Journal of Enology and Viticulture, ISSN 0002-9254, E-ISSN 1943-7749, Vol. 55, no 2, p. 199-201Article in journal (Refereed)
    Abstract [en]

    Liquefaction enzymes are often used during fermentation of fruit mashes to improve the yield of ethanol and the ability to pump the mash. The liquefaction enzymes hydrolyze pectin, but in addition to the desired pectin hydrolysis activity, the enzymes also have pectinesterase activity which hydrolyzes the methyl ester in pectin resulting in methanol formation. Fermentations of various apple varieties and Bartlett pears were conducted using liquefaction enzymes. The resulting distilled products contained concentrations of methanol above the legal limit of 280 mg/100 mL of 40% ethanol, with the observed methanol concentrations in a range between 320 and 656 mg/100 mL of 40% ethanol. In contrast to the undesired increase in methanol concentration, the enzymes provide only a marginal increase in ethanol yield. The results indicate that liquefaction enzymes should be used with caution for pomace fruit and that methanol monitoring should be implemented if these enzymes are used for commercial products.

  • 27.
    Anja, Håkansson
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Therese, Dalén
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Josefine, Gröblacher
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    William, Göransson
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Jonathan, Jaksties
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Nathalie, Ortstad
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Pauline, Lenkeit Gesser
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Biology Education Centre.
    Hello P. pastoris!: The cultivatin and expression of proteins in the yeast Pichia pastoris2022Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Producing pharmaceuticals in Escherichia coli inevitably comes with an extensive purification process. This is because many of the native proteins of E. coli are immunogenic to humans, especially the heat and pH resistant endotoxins located in the membrane of E. coli. These native proteins drive up the cost of the purification, which led to a request from the biopharmaceutical company Affibody AB. They want a review on the possibilities of producing their unique Affibody®-molecules in a new, less problematic host cell. Based on a previous bachelor project, Affibody AB chose Pichia pastoris as the candidate. P. pastoris is a methylotrophic yeast that is increasing in use when it comes to producing pharmaceuticals. In this review, multiple ways of utilizing P. pastoris are presented. The process proposals are based on 4 different promoters, pAOX1, pAOX2, pFLD1 and the pGAP. The AOX1- and AOX2-promoters and the FLD1-promoter are inducible promoters that require an inducer-molecule. An inducible promoter presents the best control of the process. The GAP-promoter is a constitutive promoter, meaning that the gene is expressed continuously. A constitutive promoter provides a process which requires fewer steps and ingredients. If the Affibody®-molecules were to be produced with P. pastoris as the host cell, the products would contain less immunogenic substances. Further, P. pastoris is also a very effective option when it comes to producing protein extracellularly. This would ultimately lead to a purification process that requires less resources.

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  • 28.
    Antonopoulou, Io
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Development of biocatalytic processes for selective antioxidant production2018Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    Feruloyl esterases (FAEs, EC 3.1.1.73) represent a subclass of carboxylic acid esterases that under normal conditions catalyze the hydrolysis of the ester bond between hydroxycinnamic acids (ferulic acid, sinapic acid, caffeic acid, p-coumaric acid) and sugar residues in plant cell walls. Based on their specificity towards monoferulates and diferulates, substitutions on the phenolic ring and on their amino acid sequence identity, they have been classified into four types (A-D) while phylogenetic analysis has resulted in classification into thirteen subfamilies (SF1-13). Under low water content, these enzymes are able to catalyze the esterification of hydroxycinnamic acids or the transesterification of their esters (donor) with alcohols or sugars (acceptor) resulting in compounds with modified lipophilicity, having a great potential for use in the tailor-made modification of natural antioxidants for cosmetic, cosmeceutical and pharmaceutical industries. The work described in this thesis focused on the selection,characterization and application of FAEs for the synthesis of bioactive esters with antioxidant activity in non-conventional media. The basis of the current classification systems was investigated in relation with the enzymes’ synthetic and hydrolytic abilities while the developed processes were evaluated for their efficiency and sustainability.

    Paper I was dedicated to the screening and evaluation of the synthetic abilities of 28 fungal FAEs using acceptors of different lipophilicity at fixed conditions in detergentless microemulsions. It was revealed that FAEs classified in phylogenetic subfamilies related to acetyl xylan esterases (SF5 and 6) showed increased transesterification rates and selectivity. In general, FAEs showed preference on more hydrophilic alcohol acceptors and in descending order to glycerol > 1-butanol > prenol. Homology modeling and small molecule docking simulations were employed as tools for the identification of a potential relationship between the predicted surface and active site properties of selected FAEs and the transesterification selectivity.

    Papers II- IV focused on the characterization of eight promising FAEs and the optimization of reaction conditions for the synthesis of two bioactive esters (prenyl ferulate and L-arabinose ferulate) in detergentless microemulsions. The effect of the medium composition, the donor and acceptor concentration, the enzyme load, the pH, the temperature and the agitation on the transesterification yield and selectivity were investigated. It was observed that the acceptor concentration and enzyme load were crucial parameters for selectivity. Fae125 (Type A, SF5) iiexhibited highest prenyl ferulate yield (81.1%) and selectivity (4.685) converting 98.5% of VFA to products after optimization at 60 mM VFA, 1.5 M prenol, 0.04 mg FAE mL-1, 40oC, 24 h, 53.4:43.4:3.2 v/v/v n-hexane: t-butanol: 100 mM MOPS-NaOH pH 8.0. On the other hand, FaeA1 (Type A, SF5) showed highest L-arabinose ferulate yield (52.2 %) and selectivity (1.120) at 80 mM VFA, 55 mM L-arabinose, 0.02 mg FAE mL-1, 50oC, 8 h, 19.8: 74.7: 5.5 v/v/v n-hexane: t-butanol: 100 mM MOPS-NaOH pH 8.0.

    In paper V, the effect of reaction media on the enzyme stability and transesterification yield and selectivity was studied in different solvents for the synthesis of two bioactive esters: prenyl ferulate and L-arabinose ferulate. The best performing enzyme (Fae125) was used in the optimization of reaction conditions in the best solvent (n-hexane) via response surface methodology. Both bioconversions were best described by a two-factor interaction model while optimal conditions were determined as the ones resulting in highest yield and selectivity.Highest prenyl ferulate yield (87.5%) and selectivity (7.616) were observed at 18.56 mM prenol mM-1VFA, 0.04 mg FAE mL-1, 24.5 oC, 24.5 h, 91.8: 8.2 v/v n-hexane: 100 mM sodium acetate pH 4.7. Highest L-arabinose ferulate yield (56.2%) and selectivity (1.284) were observed at 2.96 mM L-arabinose mM-1VFA, 0.02 mg FAE mL-1, 38.9 oC, 12 h, 90.5: 5.0: 4.5 v/v/v n-hexane: dimethyl sulfoxide: 100 mM sodium acetate pH 4.7. The enzyme could be reused for six consecutive reaction cycles maintaining 66.6% of its initial synthetic activity. The developed bioconversions showed exceptional biocatalyst productivities (> 300 g product g-1FAE) and the waste production was within the range of pharmaceutical processes.

    Paper VI focused on the investigation of the basis of the type A classification of a well-studied FAE from Aspergillus niger(AnFaeA) by comparing its activity towards methyl and arabinose hydroxycinnamic acid esters. For this purpose, L-arabinose ferulateand caffeate were synthesized enzymatically. kcat/Kmratios revealed that AnFaeA hydrolyzed arabinose ferulate 1600 times and arabinose caffeate 6.5 times more efficiently than methyl esters. This study demonstrated that short alkyl chain hydroxycinnamate esters which are used nowadays for FAE classification can lead to activity misclassification, while L-arabinose esters could potentially substitute synthetic esters in classification describing more adequately the enzyme specificitiesin the natural environment.

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  • 29.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Dilokpimol, Adiphol
    Fungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal Molecular Physiology, Utrecht University.
    Iancu, Laura
    Dupont Industrial Biosciences.
    Mäkelä, Miia R.
    Department of Microbiology, University of Helsink.
    Varriale, Simona
    Department of Chemical Sciences, University of Naples “Federico II”.
    Cerullo, Gabriella
    Department of Chemical Sciences, University of Naples “Federico II”.
    Hüttner, Silvia
    Department of Biology and Biological Engineering, Division of Industrial Biotechnology, Chalmers University of Technology.
    Uthoff, Stefan
    Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster.
    Jütten, Peter
    Taros Chemicals GmbH & Co KG.
    Piechot, Alexander
    Taros Chemicals GmbH & Co KG.
    Steinbüchel, Alexander
    nstitut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster.
    Olsson, Lisbeth
    Department of Biology and Biological Engineering, Division of Industrial Biotechnology, Chalmers University of Technology.
    Faraco, Vincenza
    Department of Chemical Sciences, University of Naples “Federico II”.
    Hildén, Kristiina
    Department of Microbiology, University of Helsinki.
    de Vries, Ronald
    Fungal Physiology, Westerdijk Fungal Biodiversity Institute & Fungal Molecular Physiology, Utrecht University.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    The Synthetic Potential of Fungal Feruloyl Esterases: A Correlation with Current Classification Systems and Predicted Structural Properties2018In: Catalysts, E-ISSN 2073-4344, Vol. 8, no 6, article id 242Article in journal (Refereed)
    Abstract [en]

    Twenty-eight fungal feruloyl esterases (FAEs) were evaluated for their synthetic abilities in a ternary system of n-hexane: t-butanol: 100 mM MOPS-NaOH pH 6.0 forming detergentless microemulsions. Five main derivatives were synthesized, namely prenyl ferulate, prenyl caffeate, butyl ferulate, glyceryl ferulate, and l-arabinose ferulate, offering, in general, higher yields when more hydrophilic alcohol substitutions were used. Acetyl xylan esterase-related FAEs belonging to phylogenetic subfamilies (SF) 5 and 6 showed increased synthetic yields among tested enzymes. In particular, it was shown that FAEs belonging to SF6 generally transesterified aliphatic alcohols more efficiently while SF5 members preferred bulkier l-arabinose. Predicted surface properties and structural characteristics were correlated with the synthetic potential of selected tannase-related, acetyl-xylan-related, and lipase-related FAEs (SF1-2, -6, -7 members) based on homology modeling and small molecular docking simulations.

  • 30.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Hunt, Cameron
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Cerullo, Gabriella
    Department of Chemical Sciences, University of Naples "Federico II".
    Varriale, Simona
    Department of Chemical Sciences, University of Naples “Federico II”.
    Gerogianni, Alexandra
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Faraco, Vincenza
    Department of Chemical Sciences, University of Naples "Federico II".
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Tailoring the specificity of the type C feruloyl esterase FoFaeC from Fusarium oxysporum towards methyl sinapate by rational redesign based on small molecule docking simulations2018In: PLOS ONE, E-ISSN 1932-6203, Vol. 13, no 5, article id e0198127Article in journal (Refereed)
    Abstract [en]

    The type C feruloyl esterase FoFaeC from Fusarium oxysporum is a newly discovered enzyme with high potential for use in the hydrolysis of lignocellulosic biomass but it shows low activity towards sinapates. In this work, small molecule docking simulations were employed in order to identify important residues for the binding of the four model methyl esters of hydroxycinnamic acids, methyl ferulate/caffeate/sinapate/p-coumarate, to the predicted structure of FoFaeC. Subsequently rational redesign was applied to the enzyme’ active site in order to improve its specificity towards methyl sinapate. A double mutation (F230H/T202V) was considered to provide hydrophobic environment for stabilization of the methoxy substitution on sinapate and a larger binding pocket. Five mutant clones and the wild type were produced in Pichia pastoris and biochemically characterized. All clones showed improved activity, substrate affinity, catalytic efficiency and turnover rate compared to the wild type against methyl sinapate, with clone P13 showing a 5-fold improvement in catalytic efficiency. Although the affinity of all mutant clones was improved against the four model substrates, the catalytic efficiency and turnover rate decreased for the substrates containing a hydroxyl substitution.

  • 31.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Iancu, Laura
    Dupont Industrial Biosciences, Wageningen, the Netherlands.
    Jütten, Peter
    Taros Chemicals GmbH & Co KG, Dortmund, Germany.
    Piechot, Alexander
    Taros Chemicals GmbH & Co KG, Dortmund, Germany.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Optimized Enzymatic Synthesis of Feruloyl Derivatives Catalyzed by Three Novel Feruloyl Esterases from Talaromyces wortmannii in Detergentless Microemulsions2018In: Computational and Structural Biotechnology Journal, E-ISSN 2001-0370, p. 361-369Article in journal (Refereed)
    Abstract [en]

    Three novel feruloyl esterases (Fae125, Fae7262 and Fae68) from Talaromyces wortmanniioverexpressed in the C1 platform were evaluated for the transesterification of vinyl ferulatewith two acceptors of different size and lipophilicity (prenol and L-arabinose) in detergentless microemulsions. The effect of reaction conditions such as the microemulsion composition, the substrate concentration, the enzyme load, the pH, the temperature and the agitation were investigated. The type A Fae125 belonging to the subfamily 5 (SF5) of phylogenetic classification showed highest yields for the synthesis of both products after optimization of reaction conditions: 81.8% for prenyl ferulate and 33.0% for L-arabinose ferulate. After optimization, an 8-fold increase in the yield and a 12-fold increase in selectivity were achieved for the synthesis of prenyl ferulate.

  • 32.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Iancu, Laura
    Dupont Industrial Biosciences.
    Jütten, Peter
    Taros Chemicals GmbH & Co KG.
    Piechot, Alexander
    Taros Chemicals GmbH & Co KG.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Screening of novel feruloyl esterases from Talaromyces wortmannii for the development of efficient and sustainable syntheses of feruloyl derivatives2019In: Enzyme and microbial technology, ISSN 0141-0229, E-ISSN 1879-0909, Vol. 120, p. 124-135Article in journal (Refereed)
    Abstract [en]

    The feruloyl esterases Fae125, Fae7262 and Fae68 from Talaromyces wortmannii were screened in 10 different solvent: buffer systems in terms of residual hydrolytic activity and of the ability for the transesterification of vinyl ferulate with prenol or L-arabinose. Among the tested enzymes, the acetyl xylan-related Fae125 belonging to the phylogenetic subfamily 5 showed highest yield and selectivity for both products in alkane: buffer systems (n-hexane or n-octane). Response surface methodology, based on a 5-level and 6-factor central composite design, revealed that the substrate molar ratio and the water content were the most significant variables for the bioconversion yield and selectivity. The effect of agitation, the possibility of DMSO addition and the increase of donor concentration were investigated. After optimization, competitive transesterification yields were obtained for prenyl ferulate (87.5-92.6%) and L-arabinose ferulate (56.2-61.7%) at reduced reaction times (≤ 24 h) resulting in good productivities (> 1 g/L/h, >300 kg product/kg FAE). The enzyme could be recycled for six consecutive cycles retaining 66.6% of the synthetic activity and 100% of the selectivity.

  • 33.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Leonov, Laura
    DuPont Industrial Biosciences.
    Jûtten, Peter
    Taros Chemicals GmbH & Co.
    Cerullo, Gabriella
    Department of Chemical Sciences, University of Naples "Federico II".
    Faraco, Vincenza
    Department of Chemical Sciences, University of Naples "Federico II".
    Papadopoulou, Adamantia
    Institute of Biosciences and Applications NCSR "Demokritos," Laboratory of Cell Proliferation and Aging.
    Kletsas, Dimitris
    Institute of Biosciences and Applications NCSR "Demokritos," Laboratory of Cell Proliferation and Aging.
    Ralli, Marianna
    Korres Natural Products.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Optimized synthesis of novel prenyl ferulate performed by feruloyl esterases from Myceliophthora thermophila in microemulsions2017In: Applied Microbiology and Biotechnology, ISSN 0175-7598, E-ISSN 1432-0614, Vol. 101, no 8, p. 3213-3226Article in journal (Refereed)
    Abstract [en]

    Five feruloyl esterases (FAEs; EC 3.1.1.73), FaeA1, FaeA2, FaeB1, and FaeB2 from Myceliophthora thermophila C1 and MtFae1a from M. thermophila ATCC 42464, were tested for their ability to catalyze the transesterification of vinyl ferulate (VFA) with prenol in detergentless microemulsions. Reaction conditions were optimized investigating parameters such as the medium composition, the substrate concentration, the enzyme load, the pH, the temperature, and agitation. FaeB2 offered the highest transesterification yield (71.5 ± 0.2%) after 24 h of incubation at 30 °C using 60 mM VFA, 1 M prenol, and 0.02 mg FAE/mL in a mixture comprising of 53.4:43.4:3.2 v/v/v n-hexane:t-butanol:100 mM MOPS-NaOH, pH 6.0. At these conditions, the competitive side hydrolysis of VFA was 4.7-fold minimized. The ability of prenyl ferulate (PFA) and its corresponding ferulic acid (FA) to scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals was significant and similar (IC50 423.39 μM for PFA, 329.9 μM for FA). PFA was not cytotoxic at 0.8–100 μM (IC50 220.23 μM) and reduced intracellular reactive oxygen species (ROS) in human skin fibroblasts at concentrations ranging between 4 and 20 μM as determined with the dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay.

  • 34.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Papadopoulou, Adamantia
    Laboratory of Cell Proliferation & Ageing, Institute of Biosciences & Applications NCSR “Demokritos”, T. Patriarchou Grigoriou & Neapoleos.
    Iancu, Laura
    DuPont Industrial Biosciences.
    Cerullo, Gabriella
    Department of Chemical Sciences, University of Naples "Federico II".
    Ralli, Marianna
    Korres Natural Products.
    Jûtten, Peter
    Taros Chemicals GmbH & Co.
    Piechot, Alexander
    Taros Chemicals GmbH & Co.
    Faraco, Vincenza
    Department of Chemical Sciences, University of Naples "Federico II".
    Kletsas, Dimitris
    Institute of Biosciences and Applications NCSR "Demokritos," Laboratory of Cell Proliferation and Aging.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Optimization of enzymatic synthesis of l-arabinose ferulate catalyzed by feruloyl esterases from Myceliophthora thermophila in detergentless microemulsions and assessment of its antioxidant and cytotoxicity activities2018In: Process Biochemistry, ISSN 1359-5113, E-ISSN 1873-3298, Vol. 65, p. 100-108Article in journal (Refereed)
    Abstract [en]

    The feruloyl esterases FaeA1, FaeA2, FaeB1, FaeB2 from Myceliophthora thermophila C1 and MtFae1a from M. thermophila ATCC 42464 were used as biocatalysts for the transesterification of vinyl ferulate (VFA) with l-arabinose in detergentless microemulsions. The effect of parameters such as the microemulsion composition, the substrate concentration, the enzyme load, the pH, the temperature and the agitation was investigated. FaeA1 offered the highest transesterification yield (52.2 ± 4.3%) after 8 h of incubation at 50 °C using 80 mM VFA, 55 mM l-arabinose and 0.02 mg FAE mL−1 in a mixture comprising of 19.8: 74.7: 5.5 v/v/v n-hexane: t-butanol: 100 mM MOPS-NaOH pH 8.0. The ability of l-arabinose ferulate (AFA) to scavenge 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals was significant (IC50 386.5 μM). AFA was not cytotoxic even at high concentrations (1 mM) however was found to be pro-oxidant at concentrations higher than 20 μM when the antioxidant activity was determined with the dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay in human skin fibroblasts.

  • 35.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Sapountzaki, Eleftheria
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Ferulic Acid From Plant Biomass: A Phytochemical With Promising Antiviral Properties2022In: Frontiers in Nutrition, E-ISSN 2296-861X, Vol. 8, article id 777576Article, review/survey (Refereed)
    Abstract [en]

    Plant biomass is a magnificent renewable resource for phytochemicals that carry bioactive properties. Ferulic acid (FA) is a hydroxycinnamic acid that is found widespread in plant cell walls, mainly esterified to polysaccharides. It is well known of its strong antioxidant activity, together with numerous properties, such as antimicrobial, anti-inflammatory and neuroprotective effects. This review article provides insights into the potential for valorization of FA as a potent antiviral agent. Its pharmacokinetic properties (absorption, metabolism, distribution and excretion) and the proposed mechanisms that are purported to provide antiviral activity are presented. Novel strategies on extraction and derivatization routes, for enhancing even further the antiviral activity of FA and potentially favor its metabolism, distribution and residence time in the human body, are discussed. These routes may lead to novel high-added value biorefinery pathways to utilize plant biomass toward the production of nutraceuticals as functional foods with attractive bioactive properties, such as enhancing immunity toward viral infections.

  • 36.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Spanopoulos, Athanasios
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, 5 Iroon Polytechniou Str, Zografou Campus, Athens, Greece.
    Matsakas, Leonidas
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Single cell oil and ethanol production by the oleaginous yeast Trichosporon fermentans utilizing dried sweet sorghum stalks2020In: Renewable energy, ISSN 0960-1481, E-ISSN 1879-0682, Vol. 146, p. 1609-1617Article in journal (Refereed)
    Abstract [en]

    The ability of the oleaginous yeast Trichosporon fermentans to efficiently produce lipids when cultivated in dried sweet sorghum was evaluated. First, lipid production was evaluated in synthetic media mimicking the composition of sweet sorghum stalks and optimized based on the nitrogen source and C: N ratio. Under optimum conditions, the lipid production reached 3.66 g/L with 21.91% w/w lipid content by using a mixture of sucrose, glucose and fructose and peptone at C: N ratio 160. Cultivation on pre-saccharified sweet sorghum stalks offered 1.97 g/L, while it was found that sweet sorghum stalks can support yeast growth and lipid production without the need for external nitrogen source addition. At an attempt to increase the carbon source concentration for optimizing lipid production, the Crabtree effect was observed in T. fermentans. To this end, the yeast was evaluated for its potential to produce ethanol under anaerobic conditions in synthetic media and sweet sorghum. The ethanol concentration at 100 g/L glucose was 40.31 g/L, while utilizing sweet sorghum by adding a distinct saccharification step and external nitrogen source offered ethanol concentration equal to 23.5 g/L. To the authors’ knowledge, this is the first time that the Crabtree effect is observed in T. fermentans.

  • 37.
    Antonopoulou, Io
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Varriale, Simona
    Department of Chemical Sciences, University of Naples "Federico II".
    Topakas, Evangelos
    National Technical University of Athens, School of Chemical Engineering, National Technical University of Athens, Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Faraco, Voncenza
    Department of Chemical Sciences, University of Naples "Federico II".
    Enzymatic synthesis of bioactive compounds with high potential for cosmeceutical application2016In: Applied Microbiology and Biotechnology, ISSN 0175-7598, E-ISSN 1432-0614, Vol. 100, no 15, p. 6519-6543Article in journal (Refereed)
    Abstract [en]

    Cosmeceuticals are cosmetic products containing biologically active ingredients purporting to offer a pharmaceutical therapeutic benefit. The active ingredients can be extracted and purified from natural sources (botanicals, herbal extracts, or animals) but can also be obtained biotechnologically by fermentation and cell cultures or by enzymatic synthesis and modification of natural compounds. A cosmeceutical ingredient should possess an attractive property such as anti-oxidant, anti-inflammatory, skin whitening, anti-aging, anti-wrinkling, or photoprotective activity, among others. During the past years, there has been an increased interest on the enzymatic synthesis of bioactive esters and glycosides based on (trans)esterification, (trans)glycosylation, or oxidation reactions. Natural bioactive compounds with exceptional theurapeutic properties and low toxicity may offer a new insight into the design and development of potent and beneficial cosmetics. This review gives an overview of the enzymatic modifications which are performed currently for the synthesis of products with attractive properties for the cosmeceutical industry

  • 38.
    Aremu, Mujidat Omolara
    et al.
    University of Borås, Faculty of Textiles, Engineering and Business.
    Ishola, M. M.
    Göteborg Energi AB, Göteborg.
    Taherzadeh, Mohammad J
    University of Borås, Faculty of Textiles, Engineering and Business.
    Polyhydroxyalkanoates (Phas) production from volatile fatty acids (vfas) from organic wastes by pseudomonas oleovorans2021In: Fermentation, ISSN 2311-5637, Vol. 7, no 4, article id 287Article in journal (Refereed)
    Abstract [en]

    This study aimed to investigate the production of polyhydroxyalkanoates (PHAs), a biodegradable polymer from organic wastes by Pseudomonas oleovorans. Volatile fatty acids (VFAs) from acidogenic fermentations of chicken manure (VFAs-CM) and potato peels (VFAs-PP), rich in organic matter majorly acetic (49.9%), butyric (15%) and propionic acids (11.1%) were utilized as substrates for microbial processes. During 72 h of cultivations, samples were withdrawn at intervals and analyzed for cell growth parameters, PHAs accumulation and polymeric properties. The highest biopolymer accumulation (0.39 g PHAs/g DCW) was achieved at 48 h of cultivation from medium containing VFAs-PP as the sole source of carbon. On characterization, the produced biopolymers were shown to be semi-crystalline of carbonyl C=O group. Additionally, thermogravimetric analysis (TGA) showed that the produced biopolymers demonstrated the capability to withstand thermal degradation above prescribed temperatures at which cross-linking isomerization reaction occurs, which is a vital property denoting the thermal stability of biopolymer. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

  • 39.
    Arnau, Laurent
    KTH, School of Chemical Science and Engineering (CHE).
    Techno-Economic Feasibility Study for the Production of Microalgae Based Plant Biostimulant2016Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    Microalgae are considered as a potential feedstock for many promising applications. Some active substances in microalgae have plant biostimulation effects potentially useful in agriculture. However, to produce such a microalgal biomass, specific microalgae cultivation and post-treatment processes must be designed to preserve active substances. A particular focus is provided on cultivation (tubular photobioreactor) and different plausible post-treatment scenarios for microalgae separation (flocculation and centrifugation) and preservation (sterilization and drying). For each step, yield and energy consumption are modeled using data taken from literature or lab and pilot scale experiments. Industrial equipment for scale-up process is also studied by comparing existing systems. These models enable to make an economic evaluation of the whole process and to study its profitability for each scenario. The breakeven price is calculated as a function of the production rate. Several parameters are suggested to improve system efficiency and profitability at the end of this study. However, a better microalgae characterization and more experiments on potential post-treatment systems are required to improve the accuracy of the model.

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  • 40.
    Arrhenius, Karine
    et al.
    RISE Research Institutes of Sweden, Materials and Production, Applied Mechanics.
    Culleton, Lucy
    NPL National Physical Laboratory, UK.
    Nwaboh, Javis
    PTB Physikalisch-Technische Bundesanstalt, Germany.
    Li, Jianrong
    VSL Van Swinden Laboratorium BV, Netherlands.
    Need for a protocol for performance evaluation of the gas analyzers used in biomethane conformity assessment2024In: Accreditation and Quality Assurance, ISSN 0949-1775, E-ISSN 1432-0517, Vol. 29, no 1, p. 69-76Article in journal (Refereed)
    Abstract [en]

    Biomethane may contain trace components that can have adverse effects on gas vehicles performances and on the pipelines when injected in the gas grid. Biomethane quality assurance against specifications is therefore crucial for the integrity of the end-users’ appliances. Analytical methods used to assess biomethane conformity assessment must be validated properly and possibly, new methods specifically for biomethane should be developed. This paper provides an overview of the biomethane quality assurance infrastructure and the challenges faced with focus on sampling, analysis methods, reference gas mixtures, and performance evaluation. Currently, requirements for analytical method validation and fit-for-purpose assessments do not exist for biomethane. The industry is in urgent need of a protocol to evaluate the fit-for-purpose of methods in a harmonized manner. Reference gas mixtures to check the accuracy of the instrument and to determine the traceability of the measurement are also urgently required. 

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  • 41.
    Aryal, Nabin
    et al.
    Biological and Chemical Engineering, Aarhus University, Aarhus N, Denmark.
    Ghimire, Nirmal
    Department of Process, Energy and Environmental Technology, University of South-Eastern Norway, Porsgrunn, Norway.
    Bajracharya, Suman
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Coupling of microbial electrosynthesis with anaerobic digestion for waste valorization2020In: Advances in Bioenergy: Volume 5 / [ed] Yebo Li; Samir Kumar Khanal, Elsevier, 2020, p. 101-127Chapter in book (Refereed)
    Abstract [en]

    Anaerobic digestion (AD) has been widely applied bioprocess to produce the biogas for fuels from organic waste degradation. AD has been integrated with other processes for increasing the digestion efficiency and waste valorization. The integration of AD with other bioprocess optimizes the production of targeted product and reduces the waste. Recently, microbial electrosynthesis (MES) was coupled with AD for the biomethane production, chemical synthesis and resource recovery. MES coupling to AD also gives an opportunity for value-added chemical generation and hence provides additional economic gains of integrated system. In MES, the remaining carbon dioxide (CO2) in biogas is reduced to methane by methanogens utilizing in situ produced hydrogen at cathode, thereby enriching methane content. Furthermore, electroactive microbes could directly accept the electron from cathode to reduce the CO2 to methane and chemicals. Therefore, CO2 fraction in the biogas could be utilized for the further chemical synthesis such as acetate, butyrate. In this chapter, advances on AD technology and MES coupling with AD are thoroughly discussed for the production of fuels and chemicals. The outputs of recent laboratory scale experiments are summarized and discussed. Furthermore, mechanism of CO2 reduction is elaborated with methane and chemical production.

  • 42.
    Atasoy, Merve
    et al.
    KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemical Engineering, Resource recovery.
    Cetecioglu, Zeynep
    KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Chemical Engineering, Resource recovery.
    Butyric acid dominant volatile fatty acids production: Bio-Augmentation of mixed culture fermentation by Clostridium butyricum2020In: Journal of Environmental Chemical Engineering, E-ISSN 2213-3437, Vol. 8, no 6, article id 104496Article in journal (Refereed)
    Abstract [en]

    The most sustainable and environmentally friendly butyric acid production method is fermentation; however, low production yield and high substrate cost limit the competition with petrol-based production. The study is aimed to enhance butyric acid production via bioaugmentated mixed culture by Clostridium butyricum. Anaerobic sequencing batch reactors (bioaugmented and control) were operated under alkali pH (pH 10) at 35 °C and fed by dairy industry wastewater as substrate. The performance of bioaugmentation was monitored in three stages: before the application, during the application (C. butyricum was injected as %10 of active reactor volume on a daily basis for seven days), after bioaugmentation. The VFA concentration and composition (by GC-FID) with the copy gene number of C. butyricum (by Q-PCR) were monitored in the bioaugmented reactor during the operation. The bioaugmentation of C. butyricum increased butyric acid production (mgCOD L-1) from 260 ± 36 to 2889 ± 180. The total VFA production (mgCOD L-1) was increased from 1434 ± 217 to 4642 ± 1778 in control and bioaugmented reactors, respectively. There was a positive correlation between the gene copies of C. butyricum with butyric, hexanoic, n-heptanoic, valeric acids production. Furthermore, the bioaugmented mixed culture had better performance than pure culture regarding butyric acid production. The cycle analysis showed that the similar butyric acid production efficiency would be obtained in the first 6 h in the bioaugmented reactor, in the first 14 h in the control reactor of the cycle. The study provides a fundamental solution to step forward to achieve next-generation biorefineries by using both monocultures modularity and mixed culture robustness and stability regarding.

  • 43.
    Bachinger, Thomas
    Linköping University, Department of Physics, Measurement Technology, Biology and Chemistry. Linköping University, The Institute of Technology.
    Multisensor arrays: for bioprocess monitoring1999Doctoral thesis, monograph (Other academic)
    Abstract [en]

    Bioprocess engineering, the technology that is focused on the exploitation of the metabolic potential of biological agents, has attracted growing interest throughout the past 50 years from both scientific and industrial communities. The products that have been brought to market range from pharmaceuticals and enzymes to food products and vitamins. The quality of human life has been improved through these efforts in many ways.

    Despite a strong research effort and the fact that microbial transformations often reach yields close to the theoretical maximum. many bioprocesses still operate at relatively low yields. One of the obstacles in effective operation is the extraction of useful information from the bioprocess. Sensors that acquire real-time information about the cells' state and their interaction with the environment in the bioreactor are seldom available. Hence, the implementation of sophisticated process control is prevented.

    In this thesis a new approach of non-invasive on-line bioprocess monitoring is evaluated. Chemical multisensor arrays (i.e. electronic noses) are used to extract information from the composition of volatiles emitted from the cell culture. The focus is on two specific areas: (i) monitoring of key variables in the bioreactor environment and (ii) monitoring of cell states and physiological events. The overall concern is, besides the increase of yield and reproducibility, the safety operation of bioprocesses.

    To cover a comprehensive area of modern bioprocessing, several organisms are investigated under different modes of operation in laboratory- and production scale processes. In repeated batch cultivations of recombinant Escherichia coli it is shown that an electronic nose can monitor biomass and specific growth rate with high accuracy. Glucose and ethanol concentration are monitored in batch cultivations of Saccharomyces cerevisiae. Bioproduct monitoring is presented in production-scale mammalian cell cultivation. The concentration of a therapeutic protein is monitored on-line in this long-term bioprocess thereby also outlining the stability of the sensor technique.

    In production-scale mammalian cell culture it is possible to follow cell transition states and monitor the reproducibility of the process. The physiological state of the cell population is revealed in laboratory-scale cultivations. It is shown that microbial contamination can be identified earlier than with conventional methods. Finally, the metabolic burden imposed on bacterial cells through strong overexpression of recombinant protein is monitored in fed-batch cultivation.

  • 44.
    Bajracharya, Suman
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Krige, Adolf
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Matsakas, Leonidas
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Advances in cathode designs and reactor configurations of microbial electrosynthesis systems to facilitate gas electro-fermentation2022In: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 354, article id 127178Article in journal (Refereed)
    Abstract [en]

    In gas fermentation, a range of chemolithoautotrophs fix single-carbon (C1) gases (CO2 and CO) when H2 or other reductants are available. Microbial electrosynthesis (MES) enables CO2 reduction by generating H2 or reducing equivalents with the sole input of renewable electricity. A combined approach as gas electro-fermentation is attractive for the sustainable production of biofuels and biochemicals utilizing C1 gases. Various platform compounds such as acetate, butyrate, caproate, ethanol, butanol and bioplastics can be produced. However, technological challenges pertaining to the microbe-material interactions such as poor gas-liquid mass transfer, low biomass and biofilm coverage on cathode, low productivities still exist. We are presenting a review on latest developments in MES focusing on the configuration and design of cathodes that can address the challenges and support the gas electro-fermentation. Overall, the opportunities for advancing CO and CO2-based biochemicals and biofuels production in MES with suitable cathode/reactor design are prospected.

  • 45.
    Bajracharya, Suman
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Krige, Adolf
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Matsakas, Leonidas
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Dual cathode configuration and headspace gas recirculation for enhancing microbial electrosynthesis using Sporomusa ovata2022In: Chemosphere, ISSN 0045-6535, E-ISSN 1879-1298, Vol. 287, Part 3, article id 132188Article in journal (Refereed)
    Abstract [en]

    High-rate production of acetate and other value-added products from the reduction of CO2 in microbial electrosynthesis (MES) using acetogens can be achieved with high reducing power where H2 appears as a key electron mediator. H2 evolution using metal cathodes can enhance the availability of H2 to support high-rate microbial reduction of CO2. Due to the low solubility of H2, the availability of H2 remains limited to the bacteria. In this study, we investigated the performances of Sporomusa ovata for CO2 reduction when dual cathodes were used together in an MES, one was regular carbon cathode, and the other was a titanium mesh that allows higher hydrogen evolution. The dual cathode configuration was investigated in two sets of MES, one set had the usual S. ovata inoculated graphite rod, and another set had a synthetic biofilm-imprinted carbon cloth. Additionally, the headspace gas in MES was recirculated to increase the H2 availability to the bacteria in suspension. High-rate CO2 reduction was observed at −0.9 V vs Ag/AgCl with dual cathode configuration as compared to single cathodes. High titers of acetate (up to ∼11 g/L) with maximum instantaneous rates of 0.68–0.7 g/L/d at −0.9 V vs Ag/AgCl were observed, which are higher than the production rates reported in literatures for S. ovata using MES with surface modified cathodes. A high H2 availability supported the high-rate acetate production from CO2 with diminished electricity input.

  • 46.
    Bajracharya, Suman
    et al.
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Krige, Adolf
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Matsakas, Leonidas
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Christakopoulos, Paul
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Microbial Electrosynthesis Using 3D Bioprinting of Sporomusa ovata on Copper, Stainless-Steel, and Titanium Cathodes for CO2 Reduction2023In: Fermentation, E-ISSN 2311-5637, Vol. 10, no 1, article id 34Article in journal (Refereed)
    Abstract [en]

    Acetate can be produced from carbon dioxide (CO2) and electricity using bacteria at the cathode of microbial electrosynthesis (MES). This process relies on electrolytically-produced hydrogen (H2). However, the low solubility of H2 can limit the process. Using metal cathodes to generate H2 at a high rate can improve MES. Immobilizing bacteria on the metal cathode can further proliferate the H2 availability to the bacteria. In this study, we investigated the performances of 3D bioprinting of Sporomusa ovata on three metal meshes—copper (Cu), stainless steel (SS), and titanium (Ti), when used individually as a cathode in MES. Bacterial cells were immobilized on the metal using a 3D bioprinter with alginate hydrogel ink. The bioprinted Ti mesh exhibited higher acetate production (53 ± 19 g/m2/d) at −0.8 V vs. Ag/AgCl as compared to other metal cathodes. More than 9 g/L of acetate was achieved with bioprinted Ti, and the least amount was obtained with bioprinted Cu. Although all three metals are known for catalyzing H2 evolution, the lower biocompatibility and chemical stability of Cu hampered its performance. Stable and biocompatible Ti supported the bioprinted S. ovata effectively. Bioprinting of synthetic biofilm on H2-evolving metal cathodes can provide high-performing and robust biocathodes for further application of MES.

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  • 47.
    Bansal, Namita
    et al.
    DOE-Great Lakes Bioenergy Research Center, Michigan State University, East Lansing.
    Bhalla, Aditya
    DOE-Great Lakes Bioenergy Research Center, Michigan State University, East Lansing.
    Pattathil, Sivakumar
    University of Georgia, Complex Carbohydrate Research Center, University of Georgia, Athens, GA.
    Adelman, Sara L.
    DOE-Great Lakes Bioenergy Research Center, Michigan State University, East Lansing.
    Hahn, Michael G
    University of Georgia, Complex Carbohydrate Research Center, University of Georgia, Athens, GA.
    Hodge, David
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Sustainable Process Engineering.
    Hegg, Eric L.
    Michigan State University, DOE-Great Lakes Bioenergy Research Center, University of Wisconsin, Madison.
    Cell wall-associated transition metals improve alkaline-oxidative pretreatment in diverse hardwoods2016In: Green Chemistry, ISSN 1463-9262, E-ISSN 1463-9270, Vol. 18, no 5, p. 1405-1415Article in journal (Refereed)
    Abstract [en]

    The responses of four diverse hardwoods (hybrid poplar, silver birch, hybrid aspen, and sugar maple) to alkaline hydrogen peroxide (AHP) pretreated at ambient temperature and pressure were analyzed to gain a deeper understanding of the cell wall properties that contribute to differences in enzymatic hydrolysis efficacy following alkaline-oxidative pretreatment. The enzymatic hydrolysis yields of these diverse hardwoods increased significantly with increasing the cell wall-associated, redox-active transition metal content. These increases in hydrolysis yields were directly correlated with improved delignification. Furthermore, we demonstrated that these improvements in hydrolysis yields could be achieved either through elevated levels of naturally-occurring metals, namely Cu, Fe, and Mn, or by the addition of a homogeneous transition metal catalyst (e.g. Cu 2,2′-bipyridine complexes) capable of penetrating into the cell wall matrix. Removal of naturally-occurring cell wall-associated transition metals by chelation resulted in substantial decreases in the hydrolysis yields following AHP pretreatment, while re-addition of metals in the form of Cu 2,2′-bipyridine complexes and to a limited extent Fe 2,2′-bipyridine complexes prior to pretreatment restored the improved hydrolysis yields. Glycome profiles showed improved extractability of xylan, xyloglucan, and pectin epitopes with increasing hydrolysis yields for the diverse hardwoods subjected to the alkaline-oxidative pretreatment, demonstrating that the strength of association between cell wall matrix polymers decreased as a consequence of improved delignification

  • 48.
    Bauer, Fredric
    et al.
    Lund University.
    Berglund, Kris
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Hulteberg, Christian
    Lund University.
    Lundgren, Joakim
    Luleå University of Technology, Department of Engineering Sciences and Mathematics, Energy Science.
    Mesfun, Sennai
    Luleå University of Technology, Department of Engineering Sciences and Mathematics, Energy Science.
    Nilsson, Robert
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Rova, Ulrika
    Luleå University of Technology, Department of Civil, Environmental and Natural Resources Engineering, Chemical Engineering.
    Wännström, Sune
    SP Technical Research Institute of Sweden.
    Comparative system analysis of carbon preserving fermentations for biofuels production2013Report (Refereed)
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  • 49.
    Beach, Elisabeth
    University of Borås, Faculty of Textiles, Engineering and Business.
    Using biological conversion to increase the value of short-chain fatty acids by mixed cultures2023Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    The development of anaerobic digestion processes has gained recognition for its potential for producing volatile fatty acids alongside bio-alcohols. In addition, the fermentation process and the products produced from biological fermentation have the possibility to overcome the abundance of organic waste in our society, which is potentially immensely rich in untapped valuable potential products. The current project aims at producing medium chained volatile fatty acids and alcohols from short-chain volatile fatty acids using hydrogen as an electron donor. For efficient conversion, inhibition of methanogenic microorganisms was performed by thermally pre-treating the microorganisms at 90 °C for 15 min. The highest observed concentrations of volatile fatty acid were 6.42 ± 0.09 g/L and the concentration of ethanol was 0.33±0.03 g/L. These concentrations were obtained with the addition of 4 g/L of liquid substrate and gas co-substrate (H2:CO2). Moreover, the predominant product from the present experiment was valeric acid and it reached its highest concentration of 1.41 g/L after 37 days. Interestingly, the addition of H2:CO2 co-substrate showed that this fermentation can be used for carbon capture and utilisation alongside hydrogen consumption in a ratio of 1:1 to increase the value of short-chain fatty acids. Furthermore, this can contribute to decreased CO2 emissions and reduced use of fossil resources for alcohol production which is in line with the global environmental goals.

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    fulltext
  • 50.
    Behravan, Gity
    et al.
    Umeå universitet.
    Sen, Srikanta
    Stockholms Universitet.
    Rova, Ulrika
    Thelander, Lars
    Umeå universitet.
    Eckstein, Fritz
    Max-Planck-Institut fur Experimentelle Medizin.
    Gräslund, Astrid
    Stockholms Universitet.
    Formation of a free radical of the sulfenylimine type in the mouse ribonucleotide reductase reaction with 2'-azido-2'-deoxycytidine 5'-diphosphate1995In: Biochimica et Biophysica Acta, Gene Structure and Expression, ISSN 0167-4781, E-ISSN 1879-2634, Vol. 1264, no 3, p. 323-329Article in journal (Refereed)
    Abstract [en]

    Mouse and Escherichia coli ribonucleotide reductases (RR) both belong to the same class of RR, where the enzyme consists of two non-identical subunits, proteins R1 and R2. A transient free radical was observed by EPR spectroscopy in the mouse RR reaction with the suicidal inhibitor 2′-azido-2′-deoxycytidine 5′-diphosphate. The detailed hyperfine structure of the EPR spectrum of the transient radical is somewhat different for the mouse and previously studied E. coli enzymes. When the positive allosteric effector ATP was replaced by the negative effector dATP, no transient radical was observed, showing that ‘normal' binding of the inhibitor to the substrate binding site is required. Using the mouse protein R2 mutants W 103Y and D266A, where the mutations have been shown to specifically block long range electron transfer between the active site of the R1 protein to the iron/radical site in protein R2, no evidence of transient radical was found. Taken together, the data suggest that the radical is located at the active site in protein R1, and is probably of the sulfenylimine type

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