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  • 51.
    Chakraborty, Reena
    et al.
    Michigan State University.
    Berglund, Kris
    Steady state fluorescence spectroscopy of pyranine as a trace extrinsic probe to study structure in aqueous sugar solutions1992Inngår i: Journal of Crystal Growth, ISSN 0022-0248, E-ISSN 1873-5002, Vol. 125, nr 1-2, s. 81-96Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The steady state fluorescence behavior of pyranine has been monitored in aqueous solutions of sucrose, glucose, fructose, and lactose. In each case it is possible to use the ratio of peak intensities at 440 and 511 nm (the PIR) as an indicator of the degree of supersaturation of the sugar solutions. We observed both bulk water and water of solvation in all solutions examined, regardless of degree of sapersaturation. In each case the amounts of bulk water and water of solvation per molecule of sugar appear to be nearly equal at saturation. We did not observe any evidence of solvent exclusive clusters in any of the systems we studied

  • 52.
    Chakraborty, Reena
    et al.
    Michigan State University.
    Berglund, Kris
    Use of pyranine as a trace fluorescent probe to study structure in aqueous sucrose solutions1991Inngår i: Particle design via crystallization / [ed] Ramu Ramanarayanan, Washington, DC: American Institute of Chemical Engineers, 1991, s. 114-123Konferansepaper (Fagfellevurdert)
    Abstract [en]

    Emission spectroscopy using pyranine as a trace fluorescent probe was evaluated as a means of studying solution structure in aqueous sucrose solutions. Spectra of pyranine doped sucrose solutions of 0-70 wt% were obtained, indicating two microenvironments. The peak intensity ratio (defined as the ratio of peak intensities at 440 nm and 511 nm) was observed to be a very sensitive function of the water content in supersaturated solutions. The two microenvironments were designated as bulk water and that near the solvated sucrose molecules. Calculations yielded six water molecules of solvation per molecule of sucrose. This is in agreement with values in literature. The number of molecules of water per molecule of sucrose in the bulk water environment varies exponentially from 165 in a 10% solution to three for a 70% solution at 20 degrees C. These results indicate that this technique can be used successfully to measure water content in supersaturated solutions.

  • 53.
    Charavgi, Maria-Despoina
    et al.
    National Technical University of Athens.
    Dimarogona, Maria
    National Technical University of Athens.
    Topakas, Evangelos
    National Technical University of Athens.
    Christakopoulos, Paul
    Chrysina, Evangelia D.
    National Hellenic Research Foundation, Athens.
    The structure of a novel glucuronoyl esterase from Myceliophthora thermophila gives new insights into its role as a potential biocatalyst2013Inngår i: Acta Crystallographica Section D: Biological Crystallography, ISSN 0907-4449, E-ISSN 1399-0047, Vol. 69, nr 1, s. 63-73Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The increasing demand for the development of efficient biocatalysts is a consequence of their broad industrial applications. Typical difficulties that are encountered during their exploitation in a variety of processes are interconnected with factors such as temperature, pH, product inhibitors etc. To eliminate these, research has been directed towards the identification of new enzymes that would comply with the required standards. To this end, the recently discovered glucuronoyl esterases (GEs) are an enigmatic family within the carbohydrate esterase (CE) family. Structures of the thermophilic StGE2 esterase from Myceliophthora thermophila (synonym Sporotrichum thermophile), a member of the CE15 family, and its S213A mutant were determined at 1.55 and 1.9 Å resolution, respectively. The first crystal structure of the S213A mutant in complex with a substrate analogue, methyl 4-O-methyl-[beta]-D-glucopyranuronate, was determined at 2.35 Å resolution. All of the three-dimensional protein structures have an [alpha]/[beta]-hydrolase fold with a three-layer [alpha][beta][alpha]-sandwich architecture and a Rossmann topology and comprise one molecule per asymmetric unit. These are the first crystal structures of a thermophilic GE both in an unliganded form and bound to a substrate analogue, thus unravelling the organization of the catalytic triad residues and their neighbours lining the active site. The knowledge derived offers novel insights into the key structural elements that drive the hydrolysis of glucuronic acid esters.

  • 54.
    Chotteau, Veronique
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Perfusion Processes2015Inngår i: Animal Cell Culture / [ed] Mohamed Al-Rubeai, Switzerland Springer: Springer , 2015, s. 407-443Kapittel i bok, del av antologi (Fagfellevurdert)
    Abstract [en]

    The interest for perfusion is increasing nowadays. This new focus has emerged from a synergy of a demand for disposable equipment and the availability of robust cell separation device, as well as the need for higher flexibility and lower investment cost. The cell separation devices mostly used today are based on filtration, i.e. alternating flow filtration, tangential flow filtration, spin-filter, or acceleration/gravity, i.e. inclined settler, centrifuge, acoustic settler. This paper gives an introduction to the basic concepts of perfusion and its practical implementation. It reviews the actual cell separation devices and describes the approaches used in the field to develop and optimize the perfusion processes.

  • 55.
    Chotteau, Veronique
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Diana, Rafael
    Ge Healthcare Bio-sciences Ab.
    Kaisermayer, Christian
    Ge Healthcare Bio-sciences Ab.
    Lindskog, Eva
    Ge Healthcare Bio-sciences Ab.
    Robinson, Craig
    Ge Healthcare Bio-sciences Ab.
    Rucker, Jimmie L.
    Ge Healthcare Bio-sciences Ab.
    Walsh, Kieron D.
    Ge Healthcare Bio-sciences Ab.
    Flexible bag for cultivation of cells2011Patent (Annet (populærvitenskap, debatt, mm))
    Abstract [en]

    An inflatable bioreactor bag for cell cultivation, which comprising a top and a bottom sheet of flexible material, joined together to form two end edges and two side edges, wherein one baffle or a plurality of baffles extend from the bottom sheet in a region where the shortest distance to any one of the two end edges is higher than about one fourth of the shortest distance between the two end edges.

  • 56.
    Chotteau, Veronique
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Zhang, Ye
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Clincke, Marie-Francoise
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Very High Cell Density in Perfusion of CHO Cells by ATF, TFF, Wave Bioreactor, and/or CellTank Technologies: Impact of Cell Density and Applications2014Inngår i: Continuous Processing in Pharmaceutical Manufacturing / [ed] Ganapathy Subramanian, Germany Weinheim: Wiley-VCH Verlagsgesellschaft, 2014, s. 339-356Kapittel i bok, del av antologi (Annet vitenskapelig)
  • 57.
    Chotteau, Véronique
    et al.
    KTH, Skolan för bioteknologi (BIO), Bioprocessteknik.
    Hjalmarsson, Håkan
    KTH, Skolan för elektro- och systemteknik (EES), Reglerteknik. KTH, Skolan för elektro- och systemteknik (EES), Centra, ACCESS Linnaeus Centre.
    Tuning of dissolved oxygen and pH PID control parameters in large scale bioreactor by lag control2012Inngår i: Proceedings of the 21st Annual Meeting of the European Society for Animal Cell Technology (ESACT), 2012, s. 327-330Konferansepaper (Fagfellevurdert)
    Abstract [en]

    A method has been developed to tune the DO and pH controller PID parameters for pilot / large scale mammalian cultivation. Our approach is to identify a model of the variable to be controlled (e.g. DO, pH) and to design several possible PID controllers based on this model. The controllers were first tested in computer simulations, followed by wet simulation and finally the best controller was tested on the real process. The approach is developed for the tuning of the DO controller of a 50 L bioreactor using microbubble continuous oxygen flow. The method, called lag control here, is based on a lead lag control design using Bode analysis where the prediction part is omitted. Experiments show that the approach results in a highly satisfactory DO control. The oxygen microbubbles were almost completely consumed before reaching the liquid surface so the oxygen flow used to maintain the DO gave an excellent indication of the cellular oxygen consumption. The control system was robust against all the perturbations, i.e. cell growth, cell bleed, addition of air-saturated fresh medium, DO set point change and a second gas sparger used to strip out the carbon dioxide. This approach was also successfully used for the tuning of a 400 L bioreactor DO controller and pH controller.

  • 58.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Project: Bioconversion of Recalcitrant Polysaccharides by Novel Oxidative Biocatalysts for the production of ethanol and other novel products (SWEDISH ENERGY AGENCY)2014Annet (Annet (populærvitenskap, debatt, mm))
  • 59.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Project: Efficient bioconversion of forest biomass insoluble polyesters with a potential use in lignocellulosic feedstock biorefineries (FORMAS)2014Annet (Annet (populærvitenskap, debatt, mm))
  • 60.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Project: Optimized esterase biocatalysts for cost-effective industrial production (EU-FP7)2014Annet (Annet (populærvitenskap, debatt, mm))
  • 61.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Project: SOUnd-DRIven BIOtechnology (VR)2014Annet (Annet (populærvitenskap, debatt, mm))
  • 62.
    Christakopoulos, Paul
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Antonopoulou, Io
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    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.
    Synthesis of biological active compounds using carbohydrate esterases as biocatalysts2014Inngår i: New Biotechnology, ISSN 1871-6784, E-ISSN 1876-4347, Vol. 31, nr Supplement, s. S90-S91Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Various fungal and bacterial carbohydrate esterases represent appealing biocatalysts that have the ability not only to deconstruct plant biomass but also to modify compounds with a potential use in food, cosmetic and pharmaceutical industries. Feruloyl esterases (FAEs, E.C. 3.1.1.73) have been proved promising candidates for the enzymatic synthesis of antioxidants allowing more flexible process configurations. Among the advantages they provide are use of lower temperatures (50-60 °C) comparing to the counterpart chemical process (150οC), one step production of one product instead of mixtures and no need of by-product and catalyst residues removal in order to produce clean and high quality substances. Glucuronoyl esterase (GE) synthetic ability needs to be explored towards the production of alkyl branched glucuronic acid derivatives which are non-ionic surfactants and have good surface properties, including biodegradability. In addition, due to their tastelessness, non skin-irritation and non toxicity, these bioactive compounds find diverse uses in the cosmetic and pharmaceutical industries.Aim of this work is the development of competitive and eco-friendly bioconversions based on transesterification reactions catalyzed by FAEs and GEs, for the production of molecules with antioxidant activity, such as phenolic fatty and sugar esters. The synthesis of four biological active compounds (prenyl ferulate, prenyl caffeate, 5-O-(trans-feruloyl)-arabinofuranose, and glyceryl ferulate) was evaluated using recombinant FAEs from Myceliopthora thermophila and Fusarium oxysporum, while the synthesis of benzyl D-glucuronate and prenyl-D-glucuronate was evaluated using recombinant GEs from M. thermophila. All reactions were carried out in ternary systems of n-hexane/alcohol/water forming surfactantless microemulsions.

  • 63.
    Christakopoulos, Paul
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Rova, Ulrika
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Project: Organosolv Biomass Pretreatment for Flexible Fuel Production (Swedish Energy Agency)2016Annet (Annet (populærvitenskap, debatt, mm))
  • 64.
    Christakopoulos, Paul
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Rova, Ulrika
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Sjöblom, Magnus
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Topakas, Evangelos
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Project: BIOcatalytic Carbon Capture and Conversion of steel flue gas to liquid hydrocarbons (FORMAS)2016Annet (Annet (populærvitenskap, debatt, mm))
  • 65.
    Christakopoulos, Paul
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    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.
    Editorial note: Advances in enzymology and enzyme engineering2012Inngår i: Computational and Structural Biotechnology Journal, ISSN 2001-0370, Vol. 2, nr 3, artikkel-id e201209001Artikkel i tidsskrift (Fagfellevurdert)
  • 66.
    Claus, Michael
    et al.
    Michigan State University.
    Berglund, Kris
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Fruit brandy production by batch column distillation with reflux2005Inngår i: Journal of food process engineering, ISSN 0145-8876, E-ISSN 1745-4530, Vol. 28, nr 1, s. 53-67Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The relationship between the operating parameters of batch fruit spirits column stills with reflux and the congener (trace compounds that provide flavors and aromas) concentrations in resulting fruit spirits has not been widely studied. Congener concentrations were determined in three different collection fractions, or "cuts," during batch distillation. Acetaldehyde and ethyl acetate were found in higher concentrations in the head cut, first overhead fraction, of the distillation and have lower boiling points relative to ethanol. 1-Propanol and isoamyl alcohol (isopentanol) were present in higher concentrations in the tail cut, third or final fraction, of the distillation and have boiling points that are higher than ethanol. Methanol has a unique concentration profile as it has higher concentrations in both the head and tail cuts, but a lower concentration in the heart cut, the middle fraction which is the desired product of the distillation. Methanol was of particular interest because the distillate must adhere to governmental regulations that limit its concentration in the product. Operating-condition parameters that were studied include the number of trays used in the distillation as well as the use of a "catalytic converter," a high surface, copper-packing material thought to catalyze formation of cyanide-containing compounds allowing them to be separated from the distillate. The effect of the number of trays used in a distillation on the concentration of ethanol and the congeners, methanol, acetaldehyde, ethyl acetate, 1-propanol and isoamyl alcohol in the final distilled spirits product is presented. An additional result of acetaldehyde production at the copper surface of the catalytic converter was also discovered in the analysis of the data.

  • 67.
    Claus, Michael J.
    et al.
    Department of Chemistry, Adrian College.
    Berglund, Kris
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Defining still parameters using Chemcad batch distillation model for modeling fruit spirits distillations2009Inngår i: Journal of food process engineering, ISSN 0145-8876, E-ISSN 1745-4530, Vol. 32, nr 6, s. 881-892Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Fruit spirits production has always been and continues to be heavily dependent upon the sensory evaluation of the distillate by the distiller. Sensory fatigue can lead to many problems of analysis for the distiller during production. As such, it is desirable to have a method of analysis that is not heavily dependent upon the senses of the distiller. Modeling of the distillation process is difficult because of the number of components present in the fruit mash and distillate, as well as because of the constantly changing thermodynamic interactions present on each tray as a result of the batch distillation process. This paper identifies a method for identifying the still parameters needed to perform predictive modeling. This involves setting a distillate flow rate and reflux ratio, as well as time or volume cutoff frames of analysis. After identifying the still characteristics, it is possible to model the distillation using the CHEMCAD batch distillation program.

  • 68.
    Crowe, Jacob D.
    et al.
    Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing.
    Feringa, Nicholas
    Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing.
    Pattathil, Sivakumar
    Complex Carbohydrate Research Center, University of Georgia, Athens, GA.
    Merritt, Brian B.
    Complex Carbohydrate Research Center, University of Georgia, Athens, GA.
    Foster, Cliff E.
    DOE-Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, MI.
    Dines, Dayna
    DOE-Great Lakes Bioenergy Research Center, Michigan State University, East Lansing, MI.
    Ong, Rebecca G.
    Department of Chemical Engineering, Michigan Technological University, Houghton, MI.
    Hodge, David
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Identification of developmental stage and anatomical fraction contributions to cell wall recalcitrance in switchgrass2017Inngår i: Biotechnology for Biofuels, ISSN 1754-6834, E-ISSN 1754-6834, Vol. 10, nr 1, artikkel-id 184Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Background: Heterogeneity within herbaceous biomass can present important challenges for processing feedstocks to cellulosic biofuels. Alterations to cell wall composition and organization during plant growth represent major contributions to heterogeneity within a single species or cultivar. To address this challenge, the focus of this study was to characterize the relationship between composition and properties of the plant cell wall and cell wall response to deconstruction by NaOH pretreatment and enzymatic hydrolysis for anatomical fractions (stem internodes, leaf sheaths, and leaf blades) within switchgrass at various tissue maturities as assessed by differing internode. Results: Substantial differences in both cell wall composition and response to deconstruction were observed as a function of anatomical fraction and tissue maturity. Notably, lignin content increased with tissue maturity concurrently with decreasing ferulate content across all three anatomical fractions. Stem internodes exhibited the highest lignin content as well as the lowest hydrolysis yields, which were inversely correlated to lignin content. Confocal microscopy was used to demonstrate that removal of cell wall aromatics (i.e., lignins and hydroxycinnamates) by NaOH pretreatment was non-uniform across diverse cell types. Non-cellulosic polysaccharides were linked to differences in cell wall response to deconstruction in lower lignin fractions. Specifically, leaf sheath and leaf blade were found to have higher contents of substituted glucuronoarabinoxylans and pectic polysaccharides. Glycome profiling demonstrated that xylan and pectic polysaccharide extractability varied with stem internode maturity, with more mature internodes requiring harsher chemical extractions to remove comparable glycan abundances relative to less mature internodes. While enzymatic hydrolysis was performed on extractives-free biomass, extractible sugars (i.e., starch and sucrose) comprised a significant portion of total dry weight particularly in stem internodes, and may provide an opportunity for recovery during processing. Conclusions: Cell wall structural differences within a single plant can play a significant role in feedstock properties and have the potential to be exploited for improving biomass processability during a biorefining process. The results from this work demonstrate that cell wall lignin content, while generally exhibiting a negative correlation with enzymatic hydrolysis yields, is not the sole contributor to cell wall recalcitrance across diverse anatomical fractions within switchgrass

  • 69.
    Crowe, Jacob D.
    et al.
    Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing.
    Zarger, Rachael A.
    Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing.
    Hodge, David
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik. Department of Chemical Engineering and Materials Science, Michigan State University; DOE-Great Lakes Bioenergy Research Center, and Department of Biosystems & Agricultural Engineering, Michigan State University.
    Relating nanoscale accessibility within plant cell walls to improved enzyme hydrolysis yields in corn stover subjected to diverse pretreatments2017Inngår i: Journal of Agricultural and Food Chemistry, ISSN 0021-8561, E-ISSN 1520-5118, Vol. 65, nr 39, s. 8652-8662Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Simultaneous chemical modification and physical reorganization of plant cell walls via alkaline hydrogen peroxide or liquid hot water pretreatment can alter cell wall structural properties impacting nanoscale porosity. Nanoscale porosity was characterized using solute exclusion to assess accessible pore volumes, water retention value as a proxy for accessible water-cell walls surface area, and solute-induced cell wall swelling to measure cell wall rigidity. Key findings concluded that delignification by alkaline hydrogen peroxide pretreatment decreased cell wall rigidity and that the subsequent cell wall swelling resulted increased nanoscale porosity and improved enzyme binding and hydrolysis compared to limited swelling and increased accessible surface areas observed in liquid hot water pretreated biomass. The volume accessible to a 90 Å dextran probe within the cell wall was found to be correlated to both enzyme binding and glucose hydrolysis yields, indicating cell wall porosity is a key contributor to effective hydrolysis yields.

  • 70.
    Danielsson, Per-Erik
    Linköpings universitet, Institutionen för systemteknik. Linköpings universitet, Tekniska högskolan.
    Implementations of the Convolution Operation1982Rapport (Annet vitenskapelig)
    Abstract [en]

    The first part of this article surveys a large number of implementations of the convolution operation (which is also known as the sum-of-products, the inner product) based on a systematic exploration of index permutations. First we assume a limited amount of parallelism in the form of an adder. Next, multipliers and RAM:s are utilized. The so called distributed arithmetic follows naturally from this approach.

    The second part brings in the concept of pipelining on the bitlevel to obtain high throughput convolvers adapted for VLSI-design (systolic arrays). The serial/parallel multiplier is analyzed in a way that unravels a vast amount new variations. Even more interesting, all these new variations can be carried over to serial/parallel convolvers. These novel devices can be implemented as linear structures of identical cells where the multipliers are embedded at equidistant intervals.

  • 71.
    de Maré, L
    et al.
    Lund Institute of Technology, Sweden.
    Velut, S
    Lund Institute of Technology, Sweden.
    Ledung, Erika
    Mälardalens högskola, Institutionen för biologi och kemiteknik.
    Cimander, C
    Novozymes Biopharma AB, Lund, Sweden .
    Norrman, Bo
    Mälardalens högskola, Institutionen för biologi och kemiteknik.
    Karlsson, E
    Lund Institute of Technology, Lund, Sweden.
    Holst, O
    Lund Institute of Technology, Lund, Sweden.
    Hagander, P
    Lund Institute of Technology, Sweden.
    A cultivation technique for E. coli fed-batch cultivations operating close to the maximum oxygen transfer capacity of the reactor2005Inngår i: Biotechnology Letters, ISSN 0141-5492, Vol. 27, nr 14, s. 983-990Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    A cultivation strategy combining the advantages of temperature-limited fed-batch and probing feeding control is presented. The technique was evaluated in fed-batch cultivations with E. coli BL21(DE3) producing xylanase in a 3 liter bioreactor. A 20% increase in cell mass was achieved and the usual decrease in specific enzyme activity normally observed during the late production phase was diminished with the new technique. The method was further tested by growing E. coli W3110 in a larger bioreactor (50 l). It is a suitable cultivation technique when the O2 transfer capacity of the reactor is reached and it is desired to continue to produce the recombinant protein.

  • 72.
    Decker, Daniel
    et al.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysiologisk botanik.
    Kleczkowski, Leszek A.
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Institutionen för fysiologisk botanik.
    Substrate specificity and inhibitor sensitivity of plant UDP-sugar producing pyrophosphorylasesManuskript (preprint) (Annet vitenskapelig)
  • 73.
    Dimarogona, M
    et al.
    Institute of Organic and Pharmaceutical Chemistry, National Hellenic Research Foundation.
    Topakas, E
    School of Chemical Engineering, National Technical University of Athens.
    Christakopoulos, Paul
    Chrysina, E.D
    Institute of Organic and Pharmaceutical Chemistry, National Hellenic Research Foundation.
    The structure of a GH10 xylanase from Fusarium oxysporum reveals the presence of an extended loop on top of the catalytic cleft2012Inngår i: Acta Crystallographica Section D: Biological Crystallography, ISSN 0907-4449, E-ISSN 1399-0047, Vol. 68, nr 7, s. 735-742Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Xylanase enzymes have been the focus of considerable research in recent decades owing to their extensive use in a variety of biotechnological applications. Previous structural studies of a number of GH10 xylanases revealed that all GH10 family members have the (β/α)8-barrel fold and their catalytic site is conserved. The structure of a new GH10 xylanase from Fusarium oxysporum (FoXyn10a) was determined at 1.94 Å resolution from crystals belonging to the tetragonal space group P41212 with five molecules per asymmetric unit. Comparison of the structure of FoXyn10a with previously determined structures of GH10 family members indicated that most of the differences were located in the loop regions between the ordered secondary-structure elements of the barrel, as expected. However, alignment of FoXyn10a with sequence and structural homologues denoted an atypically long loop connecting strand β6b and helix 6 that was only present in one other GH10 xylanase, the structure of which is not known. This structural feature may be of functional importance, with potential implications in the catalytic efficiency of the enzyme.

  • 74.
    Dimarogona, Maria
    et al.
    National Technical University of Athens, Institute of Organic and Pharmaceutical Chemistry, National Hellenic Research Foundation, Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Nikolaivits, Efstratios
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Kanelli, Maria
    National Technical University of Athens, Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Sandgren, Mats
    Department of Chemistry and Biotechnology, Swedish University of Agricultural Sciences (SLU).
    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.
    Structural and functional studies of a Fusarium oxysporum cutinase with polyethylene terephthalate modification potential2015Inngår i: Biochimica et Biophysica Acta - General Subjects, ISSN 0304-4165, E-ISSN 1872-8006, Vol. 1850, nr 11, s. 2308-2317Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    BackgroundCutinases are serine hydrolases that degrade cutin, a polyester of fatty acids that is the main component of plant cuticle. These biocatalysts have recently attracted increased biotechnological interest due to their potential to modify and degrade polyethylene terephthalate (PET), as well as other synthetic polymers.MethodsA cutinase from the mesophilic fungus Fusarium oxysporum, named FoCut5a, was expressed either in the cytoplasm or periplasm of Escherichia coli BL21. Its X-ray structure was determined to 1.9 Å resolution using molecular replacement. The activity of the recombinant enzyme was tested on a variety of synthetic esters and polyester analogues.ResultsThe highest production of recombinant FoCut5a was achieved using periplasmic expression at 16οC. Its crystal structure is highly similar to previously determined Fusarium solani cutinase structure. However, a more detailed comparison of the surface properties and amino acid interactions revealed differences with potential impact on the biochemical properties of the two enzymes. FoCut5a showed maximum activity at 40οC and pH 8.0, while it was active on three p-nitrophenyl synthetic esters of aliphatic acids (C2, C4, C12), with the highest catalytic efficiency for the hydrolysis of the butyl ester. The recombinant cutinase was also found capable of hydrolyzing PET model substrates and synthetic polymers.ConclusionsThe first reported expression and crystal structure determination of a functional cutinase from the mesophilic fungus F. oxysporum with potential application in surface modification of PET synthetic polymers.General significanceFoCut5a could be used as a biocatalyst in industrial applications for the environmentally-friendly treatment of synthetic polymers.

  • 75. Dimarogona, Maria
    et al.
    Topakas, Evangelos
    Christakopoulos, Paul
    Cellulose degradation by oxidative enzymes2012Inngår i: Computational and Structural Biotechnology Journal, ISSN 2001-0370, Vol. 2, nr 3Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Enzymatic degradation of plant biomass has attracted intensive research interest for the production of economically viable biofuels. Here we present an overview of the recent findings on biocatalysts implicated in the oxidative cleavage of cellulose, including polysaccharide monooxygenases (PMOs or LPMOs which stands for lytic PMOs), cellobiose dehydrogenases (CDHs) and members of carbohydrate-binding module family 33 (CBM33). PMOs, a novel class of enzymes previously termed GH61s, boost the efficiency of common cellulases resulting in increased hydrolysis yields while lowering the protein loading needed. They act on the crystalline part of cellulose by generating oxidized and non-oxidized chain ends. An external electron donor is required for boosting the activity of PMOs. We discuss recent findings concerning their mechanism of action and identify issues and questions to be addressed in the future.

  • 76.
    Dimarogona, Maria
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Topakas, Evangelos
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Recalcitrant polysaccharide degradation by novel oxidative biocatalysts2013Inngår i: Applied Microbiology and Biotechnology, ISSN 0175-7598, E-ISSN 1432-0614, Vol. 97, nr 19, s. 8455-8465Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The classical hydrolytic mechanism for the degradation of plant polysaccharides by saprophytic microorganisms has been reconsidered after the recent landmark discovery of a new class of oxidases termed lytic polysaccharide monooxygenases (LPMOs). LPMOs are of increased biotechnological interest due to their implication in lignocellulosic biomass decomposition for the production of biofuels and high-value chemicals. They act on recalcitrant polysaccharides by a combination of hydrolytic and oxidative function, generating oxidized and non-oxidized chain ends. They are copper-dependent and require molecular oxygen and an external electron donor for their proper function. In this review, we present the recent findings concerning the mechanism of action of these oxidative enzymes and identify issues and questions to be addressed in the future

  • 77.
    Dimarogona, Maria
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Topakas, Evangelos
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Olsson, Lisbeth
    Chalmers University of Technology.
    Christakopoulos, Paul
    Lignin boosts the cellulase performance of a GH-61 enzyme from Sporotrichum thermophile2012Inngår i: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 110, s. 480-487Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    An enzyme belonging to the glycoside hydrolase family 61 from the thermophilic fungus Sporotrichum thermophile, was functionally expressed in the methylotrophic yeast Pichia pastoris under the transcriptional control of the alcohol oxidase (AOX1) promoter. The enzyme hydrolyzed barley β-glucan, carboxymethyl cellulose, lichenan, wheat arabinoxylan and birchwood xylan showing optimal activity at pH 8 and 65 °C. A 2:1 mixture of Celluclast 1.5 L and StCel61a was capable of increasing the degree of spruce conversion by 42%. The use of substrates with varying lignin content permitted the detection of a dependence of the enhancing capacity of StCel61a on the radical scavenging capacity of the different lignocellulosics. In the presence of a reductant, StCel61a boosted the efficiency of a mixture of purified cellulases (EGII, CBHI, β-GLUC) by 20%. The synergistic activity exhibited by StCel61a and its dependence on reducing substances provide guidelines for process design towards the production of economically viable bioethanol.

  • 78. Dulebohn, Joel I.
    et al.
    Haefner, Steven C.
    Berglund, Kris
    Dunbar, Kim R.
    Reversible carbon monoxide addition to sol-gel derived composite films containing a cationic rhodium(I) complex: towards the development of a new class of molecule-based carbon monoxide sensors1992Inngår i: Chemistry of Materials, ISSN 0897-4756, E-ISSN 1520-5002, Vol. 4, nr 3, s. 506-508Artikkel i tidsskrift (Fagfellevurdert)
  • 79.
    Dunuwila, D.D.
    et al.
    Michigan State University.
    Berglund, Kris
    ATR FTIR spectroscopy for in situ measurement of supersaturation1997Inngår i: Journal of Crystal Growth, ISSN 0022-0248, E-ISSN 1873-5002, Vol. 179, nr 1-2, s. 185-193Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The current contribution establishes the technical feasibility of Attenuated Total Reflection (ATR) Fourier transform infrared (FTIR) spectroscopy for the in situ measurement of supersaturation in crystallization processes. The approach was inspired by recent advancements in ATR spectroscopy by way of various light transfer systems for remote sensing and by the increasing availability of ATR configurations well suited for remote, in situ measurements. The feasibility of the technique was investigated using a DIPPER-210® immersion probe manufactured by Axiom Analytical, Inc. Initial experiments conducted using aqueous maleic acid proved that ATR FTIR spectroscopy can be successfully employed to measure supersaturation, solubility and the metastable limit, in situ, with sufficient accuracy and precision.

  • 80.
    Dunuwila, Dilum D.
    et al.
    Michigan State University.
    Carroll, Leslie B.
    Michigan State University.
    Berglund, Kris
    An investigation of the applicability of attenuated total reflection infrared spectroscopy for measurement of solubility and supersaturation of aqueous citric acid solutions1994Inngår i: Journal of Crystal Growth, ISSN 0022-0248, E-ISSN 1873-5002, Vol. 137, nr 3-4, s. 561-568Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Currently applied methods for measurement of solubility and supersaturation based on viscometry, refractometry, interferometry and density require the separation of phases prior to measurement. ATR (attenuated total reflection) infrared spectroscopy provides a unique configuration in which the infrared spectrum of a liquid phase can be obtained in a slurry without phase separation. The applicability of the technique was investigated using a micro Circle® open boat cell equipped with a ZnSe (zinc selenide) ATR rod. Experiments conducted with aqueous citric acid proved that ATR infrared spectroscopy can be successfully employed to determine solubility and supersaturation.

  • 81.
    Dunuwila, Dilum D.
    et al.
    Michigan State University.
    Gagliardi, Charles D.
    Michigan State University.
    Berglund, Kris
    Application of controlled hydrolysis of titanium(IV) isopropoxide to produce sol-gel-derived thin films1994Inngår i: Chemistry of Materials, ISSN 0897-4756, E-ISSN 1520-5002, Vol. 6, nr 9, s. 1556-1562Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    A variety of porous optically transparent titanium carboxylate films that can be supported on silicate glass substrates have been developed. These films were made by spin casting polymeric solutions of titanium carboxylates processed using sol-gel techniques. The stability and the quality of these films have been qualitatively correlated to the extent of control of hydrolysis exerted by carboxylate modification of the precursor titanium(IV) isopropoxide. A series of carboxylic acids ranging from acetic acid to myristic acid has been used, and their effects on film quality and stability are presented.

  • 82.
    Dunuwila, Dilum D.
    et al.
    Michigan State University.
    Torgerson, B. A.
    Michigan State University.
    Chang, C. K.
    Michigan State University.
    Berglund, Kris
    Sol-gel derived titanium carboxylate thin films for optical detection of analytes1994Inngår i: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 66, nr 17, s. 2739-2744Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The feasibility of using a sol-gel derived titanium carboxylate thin film, which can be supported on glass, for the fabrication of optical test strips was investigated using a model probe/ analyte combination. The porous films, being optically transparent and baring the capability to host probe molecules, provided an excellent system to investigate the possibility of making optical test strips. A colorimetric reagent, an iron(III) porpohyrin, was chosen as the probe molecule to detect free cyanide ion concentrations in aqueous solutions. Entrapment of the porphyrin in the film was accomplished by direct dissolution of the porphyrin in the sol-gel solutions. Chemically induced structural modifications of the polymer were carried out to stabilize the encapsulated metalloporphyrin within the sol-gel derived matrix. The syntheses of this model analyte detection system and its response are presented. An optical parameter reflective of the chemical changes that occur in the system was selected as the measurement tool; its response asymptotically increased over the cyanide ion concentration range of 40-25 000 ppm.

  • 83.
    Elankovan, P.
    et al.
    Michigan State University.
    Berglund, Kris
    Contact nucleation from aqueous dextrose solutions1987Inngår i: AIChE Journal, ISSN 0001-1541, E-ISSN 1547-5905, Vol. 33, nr 11, s. 1844-1849Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Contact nucleation studies were performed with the dextrose-water system. Under some conditions two crystallographic phases were formed, which were identified in situ using a laser Raman microprobe. In addition, the growth of the contact nuclei was monitored in situ using photomicroscopy. The results are discussed in the context of contact nucleation and growth models.

  • 84.
    Elankovan, P.
    et al.
    Michigan State University.
    Berglund, Kris
    Technique for obtaining Raman spectra of contact nuclei In Situ1986Inngår i: Applied Spectroscopy, ISSN 0003-7028, E-ISSN 1943-3530, Vol. 40, nr 5, s. 712-714Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Secondary nucleation, the formation of new crystals due to the prior presence of other growing crystals, is the primary source of new particles in most industrial crystallizers. Of the various types of secondary nucleation possible, contact nucleation, wherein a disturbance of a growing crystal surface results in nuclei, is thought to be the most prevalent.

  • 85.
    El-Gamal, Rehab
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Abdel-Maksoud, Gomaa
    Cairo University, Faculty of Archaeology, Conservation Department, Giza.
    Darwish, Sawsan
    Cairo University, Faculty of Archaeology, Conservation Department, Giza.
    Topakas, Evangelos
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    FTIR analysis for the evaluation of some triazole fungicides for the treatment of wooden artifacts2018Inngår i: Mediterranean Archaeology & Archaeometry, ISSN 1108-9628, Vol. 18, nr 2, s. 141-151Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The Growth of fungi on wooden artifacts is accompanied as a rule by various physic-chemical processes making wood rigid, brittle and deformed. Therefore it is necessary to evaluate some fungicides for the preservation of wooden artifacts in order to eliminate any deformation caused by microorganisms. This study represents an attempt to use some triazole fungicides with different concentrations (propiconazole and tebuconazole) in order to assess the chemical stability of wood damaged by fungi. Fungal ageing over different periods of time was applied by using three species of fungi (Aspergillus niger, Aspergillus flavus and Penicillium chrysogenum), which were collected from different historical buildings in Egypt (such as The Mosque of Sabiile and Koutab Suleiman Agha Selehdar dated back to 1837-1839 AD, The Mosque of El Mouayed Sheikh Al-Mahmoudi dated back to 1415 to 1421 AD etc.), and were identified in previous work. Fourier transform infrared spectroscopy (FTIR) was used to evaluate the fungicides used. The results revealed that it is unfavorable to use propiconazole in the treatment of wood infested by Aspergillus flavus. However, tebuconazole can be used safely to treat wood infested by this fungus. The results also proved that increasing propiconazole and tebuconazole concentrations was needed to achieve acceptable protection against Aspergillus niger. In case of Penicillium chrysogenum, it is noticed from the results that the increased in propiconazole and tebuconazole concentrations is not recommended for treatment and a low concentration (0.25 %) is sufficient to inhibit the fungal effect.

  • 86.
    El-Gamal, Rehab
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Nikolaivits, Efstratios
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Zervakis, Georgios I.
    Agricultural University of Athens, Laboratory of General and Agricultural Microbiology.
    Abdel-Maksoud, Gomaa
    Conservation Department, Faculty of Archaeology, Cairo University, Giza.
    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.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    The use of chitosan in protecting wooden artifacts from damage by mold fungi2016Inngår i: Electronic Journal of Biotechnology, ISSN 0717-3458, E-ISSN 0717-3458, Vol. 24, s. 70-78Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Background

    Many buildings in Egypt e.g. museums, mosques and churches, do not possess controlled environments for minimizing the risks of damage of wooden artifacts due to the growth of fungi. Fungal damage usually appears as change in wood color, appearance of stains, and sometimes deformation of wooden surfaces. In this study we focused on the effect that some fungi exert on the properties of wooden artifacts and evaluated the effectiveness of different concentrations of chitosan on their protection against damage by mold fungi.

    Results

    Samples were collected from different monuments and environments, and fungi growing on them were isolated and identified. The isolated Penicillium chrysogenum, Aspergillus flavus and A. niger strains were used for the infestation of new pitch pine samples. The results revealed that the lightness of samples infected with any of the tested fungi decreased with increasing incubation times. XRD analysis showed that the crystallinity of incubated samples treated individually with the different concentrations of chitosan was lower than the crystallinity of infected samples. The crystallinity index measured by the first and the second method decreased after the first and second months but increased after the third and fourth months. This may due to the reducing of amorphous part by enzymes or acids produced by fungi in wooden samples.

    Conclusions

    The growth of fungi on the treated wood samples decreased with increasing the concentration of chitosan. Hence, it was demonstrated that chitosan prevented fungal growth, and its use could be recommended for the protection of archaeological wooden artifacts.

    Keywords

    • archaeological wood;
    • Chitosan protection;
    • crystallinity index;
    • damage of wooden artifacts;
    • filamentous fungi;
    • FTIR;
    • Fungal damage;
    • prevention of fungal growth;
    • UV spectrophotometry;
    • wood deterioration;
    • XRD
  • 87.
    Enman, Josefine
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Fungal production and solid state chemistry of eritadenine: an integrated approach to development of an active pharmaceutical ingredient2009Doktoravhandling, med artikler (Annet vitenskapelig)
    Abstract [en]

    The present thesis demonstrates an integrated approach to the development of a potential active pharmaceutical ingredient, eritadenine, a cholesterol reducing compound originating from the shiitake mushroom (Lentinus edodes). The main areas covered in the thesis are a method for quantification of eritadenine, production of eritadenine by submerged cultivation of fungal mycelia and investigation of the influence of process parameters on mycelial growth and production, and finally solid state characterizations of eritadenine. The usage of the fungus as a source of eritadenine requires an analytical tool for quantification of the compound. An HPLC method was hence developed for identification and quantification of eritadenine, using chemically synthesized eritadenine as a reference. The amount of eritadenine in fruit bodies of selected strains of shiitake was determined and with the method developed in this study, eritadenine concentrations up to ten times higher than previously reported were detected. Since both fruit bodies and mycelia of shiitake have been shown to contain eritadenine submerged cultivation of shiitake mycelia was investigated as an alternative source for this compound. The mycelia were cultivated in various submerged conditions, both in shake flasks and in bioreactors. It was found that both the mycelia and the culture media contained eritadenine, of which the major part was detected in the culture media. While the biomass concentrations were higher in shake flasks, the eritadenine concentrations were considerably higher in the bioreactors, which were assigned to morphological variations. In an attempt to improve the mycelial growth and eritadenine production, a growth promotive substance in the form of a water extract of DDGS, a by-product from drygrind ethanol facilities, was added to the culture media. It was demonstrated that an amendment of the cultivation media with this extract caused a considerable growth promotive effect on shiitake mycelia in bioreactor cultivations, along with enhanced eritadenine production. If eritadenine will be used as a pharmaceutical agent, understanding about the solid state chemistry of the compound is required. Raman spectroscopy is a valuable technique for investigation of structural properties; hence, a Raman reference spectrum with line assignments for the solid state of synthetic eritadenine was established. To further investigate the solid state chemistry of eritadenine, its synthetic analogue was slowly crystallized from water and different ethanol concentrations, at different temperatures. Solids formed from slow cooling of either water or aqueous ethanol showed crystallinity. No polymorphism was detected, irrespective of solvent system or temperature. However, dissimilar thermal behaviours were observed, deducing crystals derived from water as dihydrates and crystals derived from aqueous ethanol as 2.5 hydrates.

  • 88.
    Enman, Josefine
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Production and quantification of eritadenine, a cholesterol reducing compound in shiitake (Lentinus edodes)2007Licentiatavhandling, med artikler (Annet vitenskapelig)
    Abstract [en]

    Cardiovascular diseases are among the main causes of death in our society and there is a strong correlation between enhanced blood cholesterol levels and the development of such diseases. The popular edible fungus, shiitake mushroom (Lentinus edodes), has been shown to produce a blood cholesterol lowering compound designated eritadenine, and the hypocholesterolemic action of this compound has been quite extensively examined in rats. Eritadenine is suggested to accelerate the removal of blood cholesterol either by stimulating tissue uptake or by inhibiting tissue release; there are no indications of this compound inhibiting the biosynthesis of cholesterol. If shiitake mushrooms are to be used as a source for a potential cholesterol reducing product, it is of great importance to determine the content of eritadenine in the mushrooms as accurately as possible. Hence, in paper I methanol extraction was used to recover as much as possible of the hypocholesterolemic agent from the fungal cells. In order to analyse the target compound, a reliable and reproducible HPLC method for separation, identification and quantification of eritadenine was developed. The amounts of eritadenine in fruit bodies of four commercially cultivated shiitake mushrooms were determined, and the mushrooms under investigation exhibited up to ten times higher levels of eritadenine (3.17-6.33 mg/g dry mushrooms) than previously reported. Not only the fruit bodies of shiitake, but also its mycelia contain eritadenine. Growing fruit bodies of shiitake is a fairly demanding and time consuming process. Hence, in search for a source of eritadenine, submerged (liquid) cultivation of shiitake mycelia could be an alternative. The reason why shiitake mushrooms synthesize eritadenine is yet not clarified; i.e. the function of this secondary metabolite and the growth conditions that favour its production are not elucidated. In addition, like other filamentous fungi, shiitake exhibits different hyphal morphologies in submerged cultures depending on cultivation conditions such as medium composition, temperature, pH, inoculum concentration, dissolved oxygen and shear. The fungal metabolism and hence production of secondary metabolites is in turn affected by the morphology, as have been shown in several studies on filamentous fungi. Submerged cultivation of shiitake mycelia offers a convenient way to change the cultivating conditions in order to improve eritadenine yield and productivity. The study in paper II focused on cultivation of mycelia at different conditions, both in shake flasks and in bioreactors, to investigate the effect of pH and stirring rate on production of eritadenine. The shiitake mycelia were found to produce eritadenine, and the compound of interest was found in both the fungal cells and the growth media. The major part (90-99%) was found in the culture medium, which offers a facilitated downstream processing if large scale production of the compound is to be conducted. The mycelial morphology in the shake flask cultures were macroscopic aggregates, pellets, and the specific productivity of eritadenine was relatively low; 6.56 mg/g dry cell weight (DCW). In the bioreactor cultivations, the mycelia grew as freely dispersed filaments, showing a higher specific productivity than in the shake flasks, ranging between 26.00- 39.58 mg/g DCW. This indicates the influence of morphology on eritadenine production. The biomass yield in shake flasks and bioreactors was in parity; 0.45 g in the shake flasks and 0.25- 0.62 g in the bioreactors. A stirring rate of 50 rpm in the bioreactors was preferable for eritadenine production, whereas for biomass production it was 250 rpm, indicating the influence of agitation on both growth and productivity. The pH did not have any major impact on growth, whereas the specific productivity in the bioreactors was higher when pH was uncontrolled than controlled at 5.7.

  • 89.
    Enman, Josefine
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Hodge, David
    Berglund, Kris
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Rova, Ulrika
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Growth promotive conditions for enhanced eritadenine production during submerged cultivation of Lentinus edodes2012Inngår i: Journal of chemical technology and biotechnology (1986), ISSN 0268-2575, E-ISSN 1097-4660, Vol. 87, nr 7, s. 903-907Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Background: Mycelium of the medicinal mushroom shiitake, Lentinus edodes, is a potential source for production of the blood cholesterol reducing compound eritadenine. To increase the mycelial biomass and in turn the production of eritadenine, a potential growth promoting substance in the form of a water extract of distillers dried grains with solubles (DDGS) was added to the culture media. Results: The hot water extract of DDGS was shown to considerably increase the growth of shiitake mycelia in bioreactor cultivations; the mycelial yield was 2-3 times higher than in the control, and the highest final biomass concentration obtained was 3.4 g L -1. Further, by using shake flask cultures as inoculums the bioreactor cultivation time could be reduced by 1 week for some of the experiments. The highest final titer of eritadenine in the present study was 25.1 mg L -1, which was about 2 times higher than in the control, and was also obtained when a water extract of DDGS was added to the culture medium. Conclusion: It was demonstrated that a water extract of DDGS promoted the growth of shiitake mycelia in bioreactor cultivations, along with enhanced eritadenine production

  • 90.
    Enman, Josefine
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Hodge, David
    Berglund, Kris
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Rova, Ulrika
    Production of the bioactive compound eritadenine by submerged cultivation of shiitake (Lentinus edodes) mycelia2008Inngår i: Journal of Agricultural and Food Chemistry, ISSN 0021-8561, E-ISSN 1520-5118, Vol. 56, nr 8, s. 2609-2612Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Fruit bodies and mycelia of shiitake mushroom (Lentinus edodes) have been shown to contain the cholesterol-reducing compound eritadenine, 2(R),3(R)-dihydroxy-4-(9-adenyl)butyric acid. In the search for a production method for eritadenine, shiitake mycelia were investigated in the present study. The mycelia were cultivated both in shake flasks and in bioreactors, to investigate the effects of pH, stirring rate, and reactor type on the production and distribution of eritadenine. Both the biomass and the culture broth were examined for their eritadenine content. In the shake flasks, the final concentration of eritadenine was 1.76 mg/L and eritadenine was equally distributed between the mycelia and the growth media. In the bioreactors, the shiitake mycelia were found to contain eritadenine in relatively low levels, whereas the majority, 90.6-98.9%, was detected in the growth media. Applying a stirring rate of 250 rpm during bioreactor cultivation resulted in the highest eritadenine concentrations: 10.23 mg/L when the pH was uncontrolled and 9.59 mg/L when the pH was controlled at 5.7. Reducing the stirring rate to 50 rpm resulted in a decreased eritadenine concentration, both at pH 5.7 (5.25 mg/L) and when pH was not controlled (5.50 mg/L). The mycelia in the shake flask cultures appeared as macroscopic aggregates, whereas mycelia cultivated in bioreactors grew more as freely dispersed filaments. This study demonstrates for the first time the extra- and intracellular distribution of eritadenine produced by shiitake mycelial culture and the influence of reactor conditions on the mycelial morphology and eritadenine concentrations.

  • 91.
    Enman, Josefine
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Patra, Anuttam
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Ramser, Kerstin
    Luleå tekniska universitet, Institutionen för system- och rymdteknik, Signaler och system.
    Rova, Ulrika
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Berglund, Kris
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Solid state characterization of sodium eritadenate2011Inngår i: American Journal of Analytical Chemistry, ISSN 2156-8251, E-ISSN 2156-8278, Vol. 2, nr 2, s. 164-173Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Knowledge of the solid state is of great importance in the development of a new active pharmaceutical ingredient, since the solid form often dictates the properties and performance of the drug. In the present study, solid state characteristics of the sodium salt of the candidate cholesterol reducing compound eritadenine, 2(R), 3(R))-dihydroxy-4-(9-adenyl)-butanoic acid, were investigated. The compound was crystallized by slow cooling from water and various aqueous ethanol solutions, at different temperatures. Further, the compound solution was subjected to lyophilization and to high vacuum drying. The resulting solids were screened for polymorphism by micro Raman spectroscopy (λex = 830 nm) and the crystallinity was investigated by X-ray powder diffraction. Further, thermal analysis was applied to study possible occurrence of solvates or hydrates. Solids obtained from slow cooling showed crystallinity, whereas rapid cooling gave rise to more amorphous solids. Analysis of difference spectra of the Raman data for solids obtained from slow cooling of solution revealed subtle differences in the structures between crystals derived from pure water and crystals derived from aqueous ethanol solutions. Finally, from the thermal analysis it was deduced that crystals obtained from pure water were stoichiometrically dihydrates whereas crystals obtained from aqueous ethanol solutions were 2.5 hydrates; this formation of different hydrates were supported by the Raman difference analysis.

  • 92.
    Enman, Josefine
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Ramser, Kerstin
    Luleå tekniska universitet, Institutionen för system- och rymdteknik, Signaler och system.
    Rova, Ulrika
    Berglund, Kris
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Raman analysis of synthetic eritadenine2008Inngår i: Journal of Raman Spectroscopy, ISSN 0377-0486, E-ISSN 1097-4555, Vol. 39, nr 10, s. 1464-1468Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Eritadenine, 2(R),3(R)-dihydroxy-4-(9-adenyl)-butyric acid, is a cholesterol-reducing compound naturally occurring in the shitake mushroom (Lentinus edodes). To identify the unknown Raman spectrum of this compound, pure synthetic eritadenine was examined and the vibrational modes were assigned by following the synthesis pathway. This was accomplished by comparing the known spectra of the starting compounds adenine and D-ribose with the spectra of a synthesis intermediate, methyl 5-(6-Aminopurin-9H-9-yl)-2,3-O-isopropylidene-5-deoxy-β-D-ribofuranoside (MAIR) and eritadenine. In the Raman spectrum of eritadenine, a distinctive vibrational mode at 773 cm-1 was detected and ascribed to vibrations in the carbon chain, ν(C--C). A Raman line that arose at 1212 cm-1, both in the Raman spectrum of MAIR and eritadenine, was also assigned to ν(C--C). Additional Raman lines detected at 1526 and at 1583 cm-1 in the Raman spectrum of MAIR and eritadenine were assigned to ν(N--C) and a deformation of the purine ring structure. In these cases the vibrational modes are due to the linkage between adenine and the ribofuranoside moiety for MAIR, and between adenine and the carbon chain for eritadenine. This link is also the cause for the disappearance of adenine specific Raman lines in the spectrum of both MAIR and eritadenine. Several vibrations observed in the spectrum of D-ribose were not observed in the Raman spectrum of eritadenine due to the absence of the ribose ring structure. In the Raman spectrum of MAIR some of the D-ribose specific Raman lines disappeared due to the introduction of methyl and isopropylidene moieties to the ribose unit. With the approach presented in this study the so far unknown Raman spectrum of eritadenine could be successfully identified and is presented here for the first time.

  • 93.
    Enman, Josefine
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Rova, Ulrika
    Berglund, Kris A.
    Quantification of the bioactive compound eritadenine in selected strains of shiitake mushroom (Lentinus edodes)2007Inngår i: Journal of Agricultural and Food Chemistry, ISSN 0021-8561, E-ISSN 1520-5118, Vol. 55, nr 4, s. 1177-1180Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Cardiovascular disease is one of the most common causes of death in the Western world, and a high level of blood cholesterol is considered a risk factor. The edible fungus, shiitake mushroom (Lentinus edodes), contains the hypocholesterolemic agent eritadenine, 2(R),3(R)-dihydroxy-4-(9-adenyl)-butyric acid. This study was conducted to quantify the amount of the cholesterol reducing agent eritadenine in shiitake mushrooms, in search of a potential natural medicine against blood cholesterol. The amounts of eritadenine in the fruit bodies of four different shiitake mushrooms, Le-1, Le-2, Le-A, and Le-B, were investigated in this study. To achieve this goal, methanol extraction was used to recover as much as possible of the hypocholesterolemic agent from the fungal cells. In addition, enzymes that degrade the fungal cell walls were also used to elucidate if the extraction could be further enhanced. To analyze the target compound, a reliable and reproducible HPLC method for separation, identification, and quantification of eritadenine was developed. The shiitake strains under investigation exhibit up to 10 times higher levels of eritadenine than previously reported for other shiitake strains. Further, pretreating the mushrooms with hydrolytic enzymes before methanol extraction resulted in an insignificant increase in the amount of eritadenine released. These results indicate the potential for delivery of therapeutic amounts of eritadenine from the ingestion of extracts or dried concentrates of shiitake mushroom strains.

  • 94.
    Faisal, Abrar
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Zarebska, Agata
    Saremi, Pardis
    Korelskiy, Danil
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser.
    Ohlin, Lindsay
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser.
    Rova, Ulrika
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser.
    Hedlund, Jonas
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser.
    Grahn, Mattias
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser.
    MFI zeolite as adsorbent for selective recovery of hydrocarbons from ABE fermentation broths2014Inngår i: Adsorption, ISSN 0929-5607, E-ISSN 1572-8757, Vol. 20, nr 2-3, s. 465-470Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    1-Butanol and butyric acid are two interesting compounds that may be produced by acetone, butanol, and ethanol fermentation using e.g. Clostridium acetobutylicum. The main drawback, restricting the commercialization potential of this process, is the toxicity of butanol for the cell culture resulting in low concentrations of this compound in the broth. To make this process economically viable, an efficient recovery process has to be developed. In this work, a hydrophobic MFI type zeolite with high silica to alumina ratio was evaluated as adsorbent for the recovery of butanol and butyric acid from model solutions. Dual component adsorption experiments revealed that both butanol and butyric acid showed a high affinity for the hydrophobic MFI zeolite when adsorbed from aqueous model solutions. Multicomponent adsorption experiments using model solutions, mimicking real fermentation broths, revealed that the adsorbent was very selective to the target compounds. Further, the adsorption of butyric and acetic acid was found to be pH dependent with high adsorption below, and low adsorption above, the respective pKa values of the acids. Thermal desorption of butanol from MFI type zeolite was also studied and a suitable desorption temperature was identified.

  • 95.
    Falcon, Javier A.
    et al.
    Michigan State University.
    Berglund, Kris
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    Monitoring of antisolvent addition crystallization with Raman spectroscopy2003Inngår i: Crystal Growth & Design, ISSN 1528-7483, E-ISSN 1528-7505, Vol. 3, nr 6, s. 947-952Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Fiber optic Raman spectroscopy was used for in situ monitoring of antisolvent addition crystallization of cortisone acetate. Raman spectral features of the solvent, the antisolvent, and the solute were monitored to measure the relative concentration of the components. Different rates of antisolvent addition were monitored and the resulting solid-state form was characterized using Raman spectroscopy and DSC. Raman spectroscopy was used to monitor dehydration during storage of the product crystals. The current study demonstrates the advantages of using Raman spectroscopy to monitor the solution phase during crystallization and simultaneously monitoring the solid-state form.

  • 96.
    Feng, Lili
    et al.
    Michigan State University.
    Berglund, Kris
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik.
    ATR-FTIR for determining optimal cooling curves for batch crystallization of succinic acid2002Inngår i: Crystal Growth & Design, ISSN 1528-7483, E-ISSN 1528-7505, Vol. 2, nr 5, s. 449-452Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    The temperature profile applied during batch cooling crystallization affects the supersaturation level, which in turn affects the crystal size distribution. It is possible, in principle, to calculate the optimal cooling profile; however, the nucleation and growth kinetics are rarely known to the degree of accuracy necessary for this calculation. The current study demonstrates an alternative approach to determination of the optimal cooling profile without any prior knowledge of kinetic data or subsequent modeling. An attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectrometer was used to monitor the supersaturation level during batch cooling crystallization. The ATR-FTIR was interfaced to a LABMAX automatic reactor system that was used in a feedback mode to control the cooling rate so that the supersaturation level remained close to the solubility throughout the cooling process. The resulting temperature profile corresponds to the optimal operating conditions for the maximum in the mean crystal size.

  • 97.
    Filatov, D.
    et al.
    Umeå universitet.
    Ingemarson, R.
    Umeå universitet.
    Johansson, E.
    Umeå universitet.
    Rova, Ulrika
    Thelander, L.
    Umeå universitet.
    Mouse ribonucleotide reductase: from genes to proteins1995Inngår i: Biochemical Society Transactions, ISSN 0300-5127, E-ISSN 1470-8752, Vol. 23, nr 4, s. 903-905Artikkel i tidsskrift (Fagfellevurdert)
  • 98.
    Gerlach, Inga
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska fakulteten.
    Operator Training Simulators towards Industrial Biotechnology2015Doktoravhandling, med artikler (Annet vitenskapelig)
    Abstract [en]

    Biotechnological processes are carried out by applying advanced and demanding process strategies. Thus, fundamental understanding and knowledge about the applied microorganism is required to achieve high productivity and profitability. Therefore, computer-based simulation has been integrated into training methodologies for supplementing and expanding academic education and industrial operator training, in order to provide more effective as well as cost- and time-efficient training.

    Operator training simulators (OTS) are computer-based training tools that represent the real laboratory or plant within an interactive, virtual environment. Trainees can learn about cause-and-effect relationships during the virtual process to improve process understanding. Also, trainees and operators get the opportunity to develop, maintain and improve their operational skills. The virtual environment allows delayed operational actions and operational faults, as they are not interfering with the real process. Acceleration of the simulation speed makes training more effective. For instance, different procedures such as start-up and shut-down can be trained in a limited period of training time. Incident and trouble-shooting training can be realized in a safe, virtual environment without exposing the operator to potential risks. So far, OTS are mainly used in military, aviation, medicine and chemical industries. However, there are only a few examples where OTS are applied in biotechnological process applications.

    The aim of this thesis is to expose the benefits of using OTS for biotechnological process applications in academic education and industrial operator training. For this purpose, the feasibility of developing and applying OTS was investigated and discussed. Three OTS for the production of bio-ethanol and recombinant protein production were developed in order to investigate the benefits and their requirements as well as to assess the training effectiveness of these. To visualize the technical system including equipment and distributed control system (DCS), graphical user interfaces (GUIs) were designed, allowing the user to interact with the simulator. Mathematical models were developed and implemented in the OTS to ensure dynamic simulation of the process, where cause-and-effect relationships are realistically described. The essential part of an OTS for biotechnological process applications is the embedded sub-model which describes the biological production system. It is important that the cell growth behaviour within a bioreactor can be  simulated with sufficient accuracy in order to ensure high training effectiveness.

    In the assessment of the training effectiveness in academic education, it was observed that the students’ capability of understanding and controlling complex biotechnological processes improved after OTS training. Students showed better performance in laboratory experiments after participating in OTS pre-training. Moreover, the development of an OTS for an industrial, large-scale bio-ethanol plant illuminates the needs of OTS in biotechnological process industries. It was shown how the conceptual design methodology can be applied in designing an OTS, based on training needs from the industrial user perspective. Also, construction of the OTS including the formulation of mathematical models, the model structure which is embedded in the OTS and the design of the GUIs is covered in this thesis. Whether the use of the OTS will become cost-effective and assure maximum transfer of training depends on long term evaluation. However, application and evaluation of the developed OTS reveal the benefits and training effectiveness of these systems. The findings of the thesis might considerably facilitate the introduction of OTS into academic education and industrial operator training for biotechnological process applications.

  • 99.
    Gerlach, Inga
    et al.
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska högskolan. University of Applied Sciences, Bremen, Germany.
    Bruening, Simone
    University of Applied Sciences, Bremen, Germany .
    Gustavsson, Robert
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska högskolan.
    Mandenius, Carl-Fredrik
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska högskolan.
    Hass, Volker C.
    University of Applied Sciences Furtwangen, Villingen-Schwenningen, Germany .
    Operator training in recombinant protein production using a structured simulator model2014Inngår i: Journal of Biotechnology, ISSN 0168-1656, E-ISSN 1873-4863, Vol. 177, s. 53-59Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Model-based operator training simulators ( OTS) could be powerful tools for virtual training of operational procedures and skills of production personnel in recombinant protein processes. The applied model should describe critical events in the bioprocess so accurately that the operators ability to observe and alertly act upon these events is trained with a high degree of efficiency. In this work is shown how this is accomplished in a structured multi-compartment model for the production of a recombinant protein in an Escherichia coli fed-batch process where in particular the induction procedure, the stress effects and overflow metabolism were highlighted. The structured model was applied on the OTS platform that virtually simulated the operational bioreactor procedures in real or accelerated time. Evaluation of training using the model-based OTS showed that trained groups of operators exhibited improved capability compared with the untrained groups when subsequently performing real laboratory scale cultivations. The results suggest that this model-based OTS may provide a valuable resource for enhancing operator skills in large scale recombinant protein manufacturing.

  • 100.
    Gerlach, Inga
    et al.
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska fakulteten.
    Hass, Volker C.
    Hochschule Furtwangen, University of Applied Sciences Furtwangen, Germany.
    Mandenius, Carl-Fredrik
    Linköpings universitet, Institutionen för fysik, kemi och biologi, Teknisk biologi. Linköpings universitet, Tekniska fakulteten.
    Conceptual Design of an Operator Training Simulator for a Bio-Ethanol Plant2015Inngår i: Processes, ISSN 2227-9717, Vol. 3, nr 3, s. 664-683Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Conceptual design methodology for the configuration and procedural training with an operating training simulator (OTS) in a large-scale plant for commercial bio-ethanol production is described. The aim of the study is to show how the methodology provides a powerful way for finding the best configuration and training structure of the OTS before constructing and implementing the software of the OTS. The OTS principle, i.e., to use a computer-based virtual representation of the real process plant intended for efficient training of process operators, has long since been applied in aviation and process industries for more efficient and flawless operations. By using the conceptual design methodology (sometimes referred to as bio-mechatronics) a variety of OTS configurations with this capacity was generated. The systematic approach of for targeting the users’ (i.e., the plant management and process operators) needs resulted in better understanding and efficiency in training of hands-on skills in operating the plant. The training included general standard operating procedures for running the plant under normal operation conditions with different starch materials, handling of typical frequent disturbances as well as acting in situations not described in the standard operation procedures and applying trouble-shooting

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