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  • 351.
    Wang, F.
    et al.
    Michigan State University.
    Wachter, J.A.
    Pharmacia Corporation, Kalamazoo, MI.
    Antosz, F.J.
    Pharmacia Corporation, Kalamazoo, MI.
    Berglund, Kris
    An investigation of solvent-mediated polymorphic transformation of progesterone using in situ Raman spectroscopy2000Inngår i: Organic Process Research & Development, ISSN 1083-6160, E-ISSN 1520-586X, Vol. 4, nr 5, s. 391-395Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Many analytical techniques, such as differential scanning calorimetry (DSC), X-ray diffraction (XRD), infrared spectroscopy (IR) and Raman spectroscopy can be used to differentiate between crystalline polymorphs of the same chemical entity. While all of these techniques are routinely applied to off-line analysis of materials, Raman spectroscopy has the advantage over these other techniques in that Raman technology currently exists for in situ monitoring of the solid-phase behavior within a mixed suspension of liquid and solid. In this work, we present our results from an in situ Raman study, demonstrating the solvent-mediated polymorphic phase transformation of progesterone. In situ Raman analysis has shown that the appearance of Form I progesterone is always preceded by the formation of Form II progesterone. Phase transformation rates were found to increase monotonically as the temperature increases, which indicates that the polymorphic system is monotropic. Form I was found to be thermodynamically more stable than Form II, while Form II was found to be kinetically favored over Form I. The results from this study are consistent with Ostwald's law of stages and lead to an in-depth understanding of the polymorphic transformation process of progesterone. The in situ monitoring capabilities of Raman spectroscopy have allowed us to define the processing parameters required to control the morphology of crystalline progesterone.

  • 352.
    Wang, Zhao
    Umeå universitet, Teknisk-naturvetenskapliga fakulteten, Kemiska institutionen.
    Recalcitrance of wood to biochemical conversion: feedstock properties, pretreatment, saccharification, and fermentability2018Doktoravhandling, med artikler (Annet vitenskapelig)
    Abstract [en]

    Lignocellulose is an inexpensive and abundant renewable resource that can be used to produce advanced biofuels, green chemicals, and other bio-based products. Pretreatment and efficient enzymatic saccharification are essential features of bioconversion of lignocellulosic biomass. The aims of the research were to achieve a better understanding of the recalcitrance of woody biomass to bioconversion, to explore different pretreatment techniques that can be used to decrease the recalcitrance of the biomass and improve the digestibility of the cellulose, and to investigate by-products of acid pretreatment that cause enzymes and microorganisms to work less efficiently.

    The recalcitrance of wood from aspen, birch, and spruce was investigated before and after acid pretreatment. Before pretreatment, birch exhibited the highest recalcitrance, which was attributed to structural factors. After pretreatment, spruce showed the highest recalcitrance, which was attributed to chemical factors, such as high lignin content. Deacetylation of hybrid aspen in planta by a CE5 acetyl xylan esterase decreased the recalcitrance, and the glucose yield of enzymatic saccharification of non-pretreated wood increased with 27%.

    Pretreatment options based on ionic liquids and steam explosion were further explored. The effects of the anionic constituents of a series of imidazolium-based ionic liquids on pretreatment of aspen and spruce were investigated. [HSO4]− was efficient only for aspen, which was attributed to acid degradation of xylan. [MeCO2]− was efficient for both aspen and spruce, which was attributed to its capability to create a disordered cell wall structure rather than to removal of lignin and hemicellulose. A comparison was made between using sulfuric acid and sulfur dioxide for pretreatment of spruce. Although sulfur dioxide resulted in a pretreatment liquid that was more inhibitory to both enzymes and yeast, it was still superior to pretreatment with sulfuric acid, a phenomenon that was attributed to the particle size of the pretreated material.

    In a comparison of microbial inhibitors in pretreatment liquids from steam explosion of spruce, formaldehyde was found to be the most important inhibitor of yeast. Enzyme inhibition by catalytically non-productive adsorption to lignins and pseudo-lignin was investigated using quantitative proteomics. The results indicate that protein adsorption to pseudo-lignin can be as extensive as adsorption to real lignin. 

  • 353.
    Williams, Daniel L.
    et al.
    Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing.
    Crowe, Jacob D.
    Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing.
    Ong, Rebecca G.
    Department of Chemical Engineering, Michigan Technological University, Houghton, MI.
    Hodge, David
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Water Sorption in Pretreated Grasses as a Predictor of Enzymatic Hydrolysis Yields2017Inngår i: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 245, s. 242-249Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    This work investigated the impact of two alkaline pretreatments, ammonia fiber expansion (AFEX) and alkaline hydrogen peroxide (AHP) delignification performed over a range of conditions on the properties of corn stover and switchgrass. Changes in feedstock properties resulting from pretreatment were subsequently compared to enzymatic hydrolysis yields to examine the relationship between enzymatic hydrolysis and cell wall properties. The pretreatments function to increase enzymatic hydrolysis yields through different mechanisms; AFEX pretreatment through lignin relocalization and some xylan solubilization and AHP primarily through lignin solubilization. An important outcome was that while changes in lignin content in AHP-delignified biomass could be clearly correlated to improved response to hydrolysis, compositional changes alone in AFEX-pretreated biomass could not explain differences in hydrolysis yields. We determined the water retention value, which characterizes the association of water with the cell wall of the pretreated biomass, can be used to predict hydrolysis yields for all pretreated biomass.

  • 354.
    Williams, Daniel L.
    et al.
    Department of Chemical Engineering and Materials Science, Michigan State University, East Lansing, MI, United States.DOE Great Lakes Bioenergy Research Center, Michigan State University, Madison, MI, United States.
    Ong, Rebecca G.
    DOE Great Lakes Bioenergy Research Center, Michigan State University, Madison, MI, United States.Department of Chemical Engineering, Michigan Technological University, Houghton, MI, United States.
    Mullet, John E.
    DOE Great Lakes Bioenergy Research Center, Michigan State University, Madison, MI, United States.Department of Biochemistry & Biophysics, Texas A&M University, College Station, TX, United States.
    Hodge, David
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik. Department of Chemical & Biological Engineering, Montana State University, Bozeman, MT, United States.
    Integration of Pretreatment With Simultaneous Counter-Current Extraction of Energy Sorghum for High-Titer Mixed Sugar Production2019Inngår i: Frontiers in Energy Research, E-ISSN 2296-598X, Vol. 6, artikkel-id 133Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Sorghum (Sorghum bicolor L. Moench) offers substantial potential as a feedstock for the production of sugar-derived biofuels and biochemical products from cell wall polysaccharides (i.e., cellulose and hemicelluloses) and water-extractable sugars (i.e., glucose, fructose, sucrose, and starch). A number of preprocessing schemes can be envisioned that involve processes such as sugar extraction, pretreatment, and densification that could be employed in decentralized, regional-scale biomass processing depots. In this work, an energy sorghum exhibiting a combination of high biomass productivity and high sugar accumulation was evaluated for its potential for integration into several potential biomass preprocessing schemes. This included counter-current extraction of water-soluble sugars followed bymild NaOH or liquid hot water pretreatment of the extracted bagasse. A novel processing scheme was investigated that could integrate with current diffuser-type extraction systems for sugar extraction. In this approach, mild NaOH pretreatment (i.e., < 90 degrees C) was performed as a counter-current extraction to yield both an extracted, pretreated bagasse and a high-concentration mixed sugar stream. Following hydrolysis of the bagasse, the combined hydrolysates derived from cellulosic sugars and extractable sugars were demonstrated to be fermentable to high ethanol titers (> 8%) at high metabolic yields without detoxification using a Saccharomyces cerevisiae strain metabolically engineered and evolved to ferment xylose.

  • 355. Xiros, C.
    et al.
    Christakopoulos, Paul
    Biotechnological potential of brewers spent grain and its recent applications2012Inngår i: Waste and Biomass Valorization, ISSN 1877-2641, E-ISSN 1877-265X, Vol. 3, nr 2, s. 213-232Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Brewers spent grain (BSG) is a by-product of the brewing process corresponding to around 85% of total by-products generated. The great number of publications over the last 5 years, on the biotechnological applications of BSG, represents the increased scientific interest on it. This critical, state of the art review aims at gathering and analysing the most recent scientific efforts on the biotechnological potential of Brewer’s spent grain and on its evaluation as a feedstock for high added value products.MethodsThe assiduous bibliographic retrospection focused on the latest scientific reports. The consideration of all relevant scientific articles was thorough and critical. The classification of the scientific efforts was made not only according to the end-products but also according to the biotechnological approach adopted.ResultsBSG has been used in a wide range of biotechnological applications such as substrate for enzymes production, as a source for value-added products (antioxidants, monosaccharides, oligosaccharides, xylitol, arabitol, bioethanol, biogas or lactic acid) or for the production of functional proteins and lipids. Its applications as a carrier in various bioprocesses have also been reported.ConclusionThe implementation of BSG’s fractionation in industrial scale seems to be the next step in BSG’s exploitation. A fractionation process which allows the exploitation of biomolecules belonging to different classes, produced from one feedstock (BSG) may be used as a pattern for the implementation of the biorefinery concept in industrial scale, as long as the methods adopted ensure the functionality of the potentially valuable components.

  • 356.
    Xiros, Charilaos
    et al.
    National Technical University of Athens, Chalmers University of Technology, Department of Chemical and Biological Engineering.
    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.
    Hydrolysis and Fermentation for Cellulosic Ethanol Production2016Inngår i: Advances in Bioenergy: The Sustainability Challenge, Chichester, UK: John Wiley & Sons Ltd , 2016, s. 11-31Kapittel i bok, del av antologi (Fagfellevurdert)
  • 357.
    Xiros, Charilaos
    et al.
    Chalmers University of Technology, Department of Chemical and Biological Engineering.
    Topakas, Evangelos
    BIOtechMASS Unit, 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.
    Hydrolysis and fermentation for cellulosic ethanol production2013Inngår i: Wiley Interdisciplinary Reviews: Energy and Environment, ISSN 2041-8396, E-ISSN 2041-840X, Vol. 2, nr 6, s. 633-654Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Second-generation bioethanol produced from various lignocellulosic materials, such as wood, agricultural, or forest residues, has the potential to be a valuable substitute for, or a complement to, gasoline. At least three major factors—rapidly increasing atmospheric CO2 levels, dwindling fossil fuel reserves, and their rising costs—suggest that we now need to accelerate research plans to make greater use of plant-based biomass for energy production and as a chemical feedstock as part of a sustainable energy economy. Optimizing the production of bioethanol to be competitive with petrochemical fuels is the main challenge for the underlying process development. The exhaustive research on enzyme technology during the latest years, resulting in significant advances in the field, show the importance of the enzymatic hydrolysis for a profitable ethanol production process. On the other hand, the persisting challenges in biomass pretreatment, which are the initial steps in most process designs, show the remarkable recalcitrance of the lignocellulosic materials to biological degradation. The recent scientific trends show toward an integrated overall bioconversion process in which fermentation technology and genetic engineering of ethanologenic microorganisms aim not only at maximizing yields and productivities but also at widening the range of fermentation products and applications.

  • 358.
    Xiros, Charilaos
    et al.
    National Technical University of Athens.
    Vafiadi, Christina
    National Technical University of Athens.
    Topakas, Evangelos
    National Technical University of Athens.
    Christakopoulos, Paul
    Decrement of cellulose recalcitrance by treatment with ionic liquid (1-ethyl-3-methylimidazolium acetate) as a strategy to enhance enzymatic hydrolysis2012Inngår i: Journal of chemical technology and biotechnology (1986), ISSN 0268-2575, E-ISSN 1097-4660, Vol. 87, nr 5, s. 629-634Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    BACKGROUND: The high crystallinity of cellulose underlies the recalcitrance that this polymer presents in enzymatic degradation. Thus, a pre-treatment step is applied in most bioconversion processes. Treatments with ionic liquids are considered an emerging pre-treatment technology, owing to their high efficiency in solvating cellulose, over molecular solvent systems.RESULTS: Crystalline cellulose with and without ionic liquid (1-ethyl-3-methylimidazolium acetate) treatment, both commercially available, were used as substrates in enzymatic hydrolysis reactions using the earlier evaluated cellulolytic system of Fusarium oxysporum. The in situ removal of the hydrolysate during reactions enhanced the reaction rate as well as the overall glucose production. Ionic liquid treatment significantly decreased cellulose crystallinity and enhanced bioconversion yields and rates. The effects of cellulose structural changes during treatment on hydrolysis rate were investigated and the recalcitrance constants were determined.CONCLUSION: The study showed that ionic liquid-treated cellulose became more homogeneous and more easily degradable than the untreated cellulose, a conclusion that was expressed mathematically by the difference in the recalcitrance constants for the two substrates. It was concluded that glucose production from ionic liquid-treated cellulose could achieve very high conversion yields in consolidated bioprocesses or during simultaneous saccharification and fermentation

  • 359.
    Yedur, Sanjay K
    et al.
    Michigan State University.
    Berglund, Kris
    Use of fluorescence spectroscopy in concentration and supersaturation measurements in citric acid solutions1996Inngår i: Applied Spectroscopy, ISSN 0003-7028, E-ISSN 1943-3530, Vol. 50, nr 7, s. 866-870Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Measurement of supersaturation is of critical importance in the operation and control of crystallizers. In this work, we report a novel spectroscopic technique to achieve the measurement of concentration and supersaturation in crystallizing solutions. In order to develop a sensor for this measurement, citric acid is chosen as the model solute, and the analytical technique involves fluorescence spectroscopy. Citric acid is a common food-grade compound with a wide range of applications that is exclusively produced by crystallization. The fluorescent properties of a probe, 8-hydroxy-1,3,6-pyrenetrisulfonate (pyranine), are used to provide concentration measurements in aqueous citric acid solutions, thereby providing for supersaturation estimation. The change in the relative emission peak intensities of the probe in different solute concentrations gives an excellent calibration curve for concentration measurements. It is also shown that, although pyranine responds to both its solvent microenvironment and the pH of the solution, it is still possible to measure concentration and supersaturation by using this fluorescence technique.

  • 360.
    Yedur, Sanjay K.
    et al.
    Michigan State University.
    Dulebohn, Joel
    Grand River Technologies, Lansing, MI.
    Werpy, Todd
    Michigan Biotechnology Institute, Lansing.
    Berglund, Kris
    Synthesis and testing of catalysts for the production of maleic anhydride from a fermentation feedstock1996Inngår i: Industrial & Engineering Chemistry Research, ISSN 0888-5885, E-ISSN 1520-5045, Vol. 35, nr 3, s. 663-671Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    It is necessary to develop alternate pathways for the production of chemicals that are traditionally produced from fossil fuels to reduce our dependency on nonrenewable energy sources. In this paper, an alternate technology is presented for producing maleic anhydride from a fermentation feedstock. The process involves the catalytic oxydehydrogenation of fermentation-derived succinic anhydride to produce maleic anhydride. Various catalysts have been synthesized and tested for the oxydehydrogenation reaction. Iron phosphate based catalysts are found to be the best on the basis of high conversions and selectivities obtained. The effects of temperature, oxygen concentration, contact time, and the total time on stream on the performance of the catalyst are investigated, and an optimum set of conditions for the operation of the bench-scale reactor is presented. The bulk and surface compositions, the surface areas, and the bulk crystallographic structure of the catalysts are also reported.

  • 361.
    Yuan, Zhaoyang
    et al.
    Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, United States.
    Singh, Sandip Kumar
    Department of Chemical & Biological Engineering, Montana State University, Montana, United States.
    Bals, Bryan
    Michigan Biotechnology Institute, Lansing, Michigan, United States.
    Hodge, David B.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Industriell miljö- och processteknik. Department of Chemical & Biological Engineering, Montana State University, Bozeman, Montana , United States.
    Hegg, Eric L.
    Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan, United States.
    Integrated Two-Stage Alkaline–Oxidative Pretreatment of Hybrid Poplar. Part 2: Impact of Cu-Catalyzed Alkaline Hydrogen Peroxide Pretreatment Conditions on Process Performance and Economics2019Inngår i: Industrial & Engineering Chemistry Research, ISSN 0888-5885, E-ISSN 1520-5045, Vol. 58, nr 35, s. 16000-16008Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Two-stage alkaline/copper 2,2′-bipyridine-catalyzed alkaline hydrogen peroxide (Cu-AHP) pretreatment is an effective strategy for improving the enzymatic digestibility of hybrid poplar. To reduce the chemical inputs and processing costs associated with this process, we investigated the effect of increasing the temperature for both the alkaline pre-extraction and the Cu-AHP pretreatment stages. The results indicate that increasing the alkaline pre-extraction and the Cu-AHP pretreatment temperatures from 30 to 120 and 80 °C, respectively, allowed us to reduce both the pretreatment time of the Cu-AHP stage and the chemical loadings. Incubating alkaline pre-extracted hybrid poplar for 12 h with 10% NaOH (w/w biomass), 8% hydrogen peroxide (w/w biomass), and a Cu2+ and 2,2′-bipyridine (bpy) concentration of 1 mM yielded monomeric sugar yields of approximately 77% glucose and 66% xylose (based on the initial sugar composition) following enzymatic hydrolysis. Technoeconomic analysis (TEA) indicates that these changes to the two-stage alkaline/Cu-AHP pretreatment process could potentially reduce the minimum fuel selling price (MFSP) by more than $1.00 per gallon of biofuel compared to the reference case where both stages were conducted at 30 °C with higher chemical inputs.

  • 362.
    Zamani, Leila
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Zhang, Ye
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Aberg, Magnus
    Lindahl, Anna
    Mie, Axel
    Chotteau, Veronique
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Metabolic footprinting of CHO cell culture bioprocess data in fed-batch and perfusion mode using LC-MS data and multivariate analysisManuskript (preprint) (Annet vitenskapelig)
  • 363.
    Zerva, Anastasia
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Antonopoulou, Io
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Enman, Josefine
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Iancu, Laura
    DuPont Industrial Biosciences, Wageningen, The Netherlands.
    Jütten, Peter
    Taros Chemicals GmbH & Co. KG, Dortmund, Germany.
    Rova, Ulrika
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Optimization of Transesterification Reactions with CLEA-Immobilized Feruloyl Esterases from Thermothelomyces thermophila and Talaromyces wortmannii2018Inngår i: Molecules, ISSN 1420-3049, E-ISSN 1420-3049, Vol. 23, nr 9, artikkel-id 2403Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Feruloyl esterases (FAEs, E.C. 3.1.1.73) are biotechnologically important enzymes with several applications in ferulic acid production from biomass, but also in synthesis of hydroxycinnamic acid derivatives. The use of such biocatalysts in commercial processes can become feasible by their immobilization, providing the advantages of isolation and recycling. In this work, eight feruloyl esterases, immobilized in cross-linked enzyme aggregates (CLEAs) were tested in regard to their transesterification performance, towards the production of prenyl ferulate (PFA) and arabinose ferulate (AFA). After solvent screening, comparison with the activity of respective soluble enzymes, and operational stability tests, FAE125 was selected as the most promising biocatalyst. A central composite design revealed the optimum conditions for each transesterification product, in terms of water content, time, and substrate ratio for both products, and temperature and enzyme load additionally for prenyl ferulate. The optimum product yields obtained were 83.7% for PFA and 58.1% for AFA. FAE125 CLEAs are stable in the optimum conditions of transesterification reactions, maintaining 70% residual activity after five consecutive reactions. Overall, FAE125 CLEAs seem to be able to perform as a robust biocatalyst, offering satisfactory yields and stability, and thus showing significant potential for industrial applications.

  • 364.
    Zerva, Anastasia
    et al.
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Antonopoulou, Io
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Enman, Josefine
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Iancu, Laura
    DuPont Industrial Biosciences, Nieuwe Kanaal 7-S, 6709 PA Wageningen, The Netherlands.
    Rova, Ulrika
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Cross-Linked Enzyme Aggregates of Feruloyl Esterase Preparations from Thermothelomyces thermophila and Talaromyces wortmannii2018Inngår i: Catalysts, ISSN 2073-4344, Vol. 8, nr 5, artikkel-id 208Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Cross-linked enzyme aggregates (CLEA®) technology is a well-established method in the current literature for the low-cost and effective immobilization of several enzymes. The main advantage of this particular method is the simplicity of the process, since it consists of only two steps. However, CLEA immobilization must be carefully designed for each desired enzyme, since the optimum conditions for enzymes can vary significantly, according to their physicochemical properties. In the present study, an investigation of the optimum CLEA immobilization conditions was carried out for eight feruloyl esterase preparations. Feruloyl esterases are a very important enzyme group in the valorization of lignocellulosic biomass, since they act in a synergistic way with other enzymes for the breakdown of plant biomass. Specifically, we investigated the type and concentration of precipitant and the crosslinker concentration, for retaining optimal activity. FAE68 was found to be the most promising enzyme for CLEA immobilization, since in this case, the maximum retained activity, over 98%, was observed. Subsequently, we examined the operational stability and the stability in organic solvents for the obtained CLEA preparations, as well as their structure. Overall, our results support that the maximum activity retaining and the stability properties of the final CLEAs can vary greatly in different FAE preparations. Nevertheless, some of the examined FAEs show a significant potential for further applications in harsh industrial conditions.

  • 365.
    Zerva, Anastasia
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Topekas, Eangelos
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Characterization and application of a novel class II thermophilic peroxidase from Myceliophthora thermophila in biosynthesis of polycatechol2015Inngår i: Enzyme and microbial technology, ISSN 0141-0229, E-ISSN 1879-0909, Vol. 75-76, s. 49-56Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    A peroxidase from the thermophilic fungus Myceliophthora thermophila that belongs to ascomycete Class II based on PeroxiBase classification was functionally expressed in methylotrophic yeast Pichia pastoris. The putative peroxidase from the genomic DNA was successfully cloned in P. pastoris X-33 under the transcriptional control of the alcohol oxidase (AOX1) promoter. The heterologous production was greatly enhanced by the addition of hemin with a titer of 0.41 U mL−1 peroxidase activity at the second day of incubation. The recombinant enzyme was purified to homogeneity (50 kDa) and characterized using a series of phenolic substrates that indicated similar characteristics with those of generic peroxidases. In addition, the enzyme was found thermostable, retaining its activity for temperatures up to 60 oC after eight hours incubation. Moreover, the enzyme displayed remarkable H2O2 stability, retaining more than 80% of its initial activity after 24 hours incubation in 5000- fold molar excess of H2O2. The ability of the peroxidase to polymerize catechol at high superoxide concentrations, together with its high thermostability and substrate specificity, indicate a potential commercial significance of the enzyme.

  • 366.
    Zerva, Anastasia
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, Athens, Greece.
    Koutroufini, Efthymia
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens, Athens, Greece.
    Kostopoulou, Ioanna
    Laboratory of Organic Chemistry, School of Chemical Engineering, National Technical University of Athens, Athens, Greece.
    Detsi, Anastasia
    Laboratory of Organic Chemistry, School of Chemical Engineering, National Technical University of Athens, Athens, Greece.
    Topakas, Evangelos
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    A novel thermophilic laccase-like multicopper oxidase from Thermothelomyces thermophila and its application in the oxidative cyclization of 2′,3,4-trihydroxychalcone2019Inngår i: New Biotechnology, ISSN 1871-6784, E-ISSN 1876-4347, Vol. 49, s. 10-18Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Laccase-like multicopper oxidases (LMCOs) are a heterogeneous group of oxidases, acting mainly on phenolic compounds and which are widespread among many microorganisms, including Basidiomycetes and Ascomycetes. Here, we report the cloning, heterologous expression, purification and characterization of a novel LMCO from the thermophilic fungus Thermothelomyces thermophila. The 1953 bp lmco gene sequence comprises of 3 exons interrupted by 2 introns and according to the LccED database the translated sequence belongs to superfamily 6 of multicopper oxidases. After removal of the introns, the gene was transformed into Pichia pastoris, under the control of the alcohol oxidase (AOX1) promoter. The heterologous enzyme was purified with an apparent molecular weight of 80 kDa. TtLMCO1 displayed optimum activity at pH 4 and 50 °C and appeared thermostable up to 50 °C. A variety of phenolic compounds were oxidized by TtLMCO1, including standard laccase substrates such as ABTS and 2,6 dimethoxyphenol. The UV/Vis spectrum of purified TtLMCO1 indicates that it belongs to yellow laccase-like oxidases. The enzyme was used for the bioconversion of 2′,3,4-trihydroxychalcone to 3′,4′-dihydroxy-aurone, a bioactive aurone recently shown to possess inhibitory activity against several isoforms of the histone deacetylase complex (HDAC). Overall, the thermophilic yellow LMCO TtLMCO1 presents a number of superior properties with potential use in industrial biocatalysis.

  • 367.
    Zerva, Anastasia
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Manos, Nikolaos
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Vouyiouka, Stamatina
    National Technical University of Athens, Laboratory of Polymer Technology, School of Chemical Engineering, National Technical University of Athens.
    Christakopoulos, Paul
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Topakas, Evangelos
    Luleå tekniska universitet, Institutionen för samhällsbyggnad och naturresurser, Kemiteknik.
    Bioconversion of Biomass-Derived Phenols Catalyzed by Myceliophthora thermophila Laccase2016Inngår i: Molecules, ISSN 1420-3049, E-ISSN 1420-3049, Vol. 21, nr 5, artikkel-id 550Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Biomass-derived phenols have recently arisen as an attractive alternative for building blocks to be used in synthetic applications, due to their widespread availability as an abundant renewable resource. In the present paper, commercial laccase from the thermophilic fungus Myceliophthora thermophila was used to bioconvert phenol monomers, namely catechol, pyrogallol and gallic acid in water. The resulting products from catechol and gallic acid were polymers that were partially characterized in respect to their optical and thermal properties, and their average molecular weight was estimated via solution viscosity measurements and GPC. FT-IR and 1H-NMR data suggest that phenol monomers are connected with ether or C–C bonds depending on the starting monomer, while the achieved molecular weight of polycatechol is found higher than the corresponding poly(gallic acid). On the other hand, under the same condition, pyrogallol was dimerized in a pure red crystalline compound and its structure was confirmed by 1H-NMR as purpurogallin. The herein studied green synthesis of enzymatically synthesized phenol polymers or biological active compounds could be exploited as an alternative synthetic route targeting a variety of applications.

  • 368.
    Zerva, Anastasia
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens .
    Papaspyridi, Lefki Maria
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens .
    Christakopoulos, Paul
    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.
    Valorization of Olive Mill Wastewater for the Production of β-glucans from Selected Basidiomycetes2017Inngår i: Waste and Biomass Valorization, ISSN 1877-2641, E-ISSN 1877-265X, Vol. 8, nr 5, s. 1721-1731Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Purpose

    The aim of the present study was to investigate the feasibility of polysaccharides production by selected basidiomycetes in submerged culture. Olive mill wastewater (OMWW) was also tested as a potential substrate for polysaccharides production by mushroom strains, focusing on the simultaneous degradation and valorization of the waste material.

    Methods

    The tested strains were grown in two different substrates, and after biomass harvesting, polysaccharides were isolated using two different methods. The extracellular polysaccharides were isolated from the culture broth, with ethanol precipitation. The isolated fractions were partially characterized with FT-IR spectroscopy.

    Results

    All three strains performed well in both substrates. Maximum degradation performance of OMWW was achieved by Ganoderma lucidum, achieving 19.4% phenols reduction together with 47.56% decolorization. The extracellular polysaccharides (EPS) produced by all strains were found to be richer in total glucans during growth in semi-synthetic medium, compared to growth in OMWW-based medium. In regard to biomass polysaccharides, Pleurotus ostreatus biomass was found to be richer in glucans, reaching 8.68% (w/w) total glucan content when grown in semi-synthetic medium and 7.58% (w/w) when grown in OMWW-based medium. After purification of biomass polysaccharides with two methods, the fraction with the highest glucan content was found to be the one from G. lucidum after growth in semi-synthetic medium cultures, with 49.1% (w/w) total glucans. FT-IR spectra of the isolated samples revealed the bands corresponding to α- and β-glucosidic bonds, but also the existence of protein contamination.

    Conclusions

    Purification of biomass polysaccharides with two distinct methods revealed that α-amylase and Sevag treatments failed to remove completely α-glucans and proteins respectively, leading to the suggestion that these two steps could be omitted without significant impact. Moreover, the results imply that the valorization of OMWW might be feasible with the use of mushroom strains, leading to the production of important products, such as glucans.

  • 369.
    Zerva, Anastasia
    et al.
    Biotechnology Laboratory, School of Chemical Engineering, National Technical University of Athens.
    Zervakis, Georgios I.
    Agricultural University of Athens, Laboratory of General and Agricultural Microbiology.
    Christakopoulos, Paul
    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.
    Degradation of olive mill wastewater by the induced extracellular ligninolytic enzymes of two wood-rot fungi2017Inngår i: Journal of Environmental Management, ISSN 0301-4797, E-ISSN 1095-8630, Vol. 203:2, s. 791-798Artikkel i tidsskrift (Fagfellevurdert)
    Abstract [en]

    Olive mill wastewater (OMWW) is a major problem in olive oil – producing countries, due to its high organic load and concentration in phenols that are toxic for marine life, plants and soil microorganisms. In the present study, two mushroom species were tested in regard to their OMWW's oxidative capacity, Pleurotus citrinopileatus LGAM 28684 and Irpex lacteus LGAM 238. OMWW (25% v/v) degradation was investigated for several culture conditions, namely pH, agitation speed, nitrogen-based supplements and their concentration. The selected values were pH 6, agitation rate 150 rpm, 30 g L−1 corn steep liquor as nitrogen source for P. citrinopileatus and 20 g L−1 diammonium tartrate for I. lacteus. The two strains performed well in cultures supplemented with OMWW, generating very high titers of oxidative enzymes and achieving more than 90% color and phenols reduction within a 24 days cultivation period. In addition, the amount of glucans present in the fungal biomass was assessed. Hence, P. citrinopileatus and I. lacteus appear as potent degraders of OMWW with the ability to use the effluent as a substrate for the production of biotechnologically important enzymes and valuable fungal glucans. 

  • 370.
    Zhang, Ye
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    High cell density perfusion process development for antibody producing Chinese Hamster Ovary cells2017Doktoravhandling, med artikler (Annet vitenskapelig)
    Abstract [en]

    Perfusion operation mode is currently under fast expansion in mammalian cell based manufacturing of biopharmaceuticals, not only for labile drug protein but also for stable proteins such as monoclonal antibodies (mAbs). Perfusion mode can advantageously offer a stable cell environment, long-term production with high productivity and consistent product quality. Intensified high cell density culture (HCDC) is certainly one of the most attractive features of a perfusion process due to the high volumetric productivity in a small footprint that it can provide. Advancements in single-use technology have alleviated the intrinsic complexity of perfusion processes while the maturing in cell retention devices has improved process robustness. The knowledge for perfusion process has been gradually built and the “continuous” concept is getting more and more acceptance in the field.

    This thesis presents the development of robust perfusion process at very high cell densities in various culture systems. Four HCDC perfusion systems were developed with industrial collaborators with three different mAb producing Chinese Hamster Ovary (CHO) cell lines: 1-2) WAVE Bioreactor™ Cellbag prototype equipped with cell separation by hollow fiber filter utilizing Alternating Tangential Flow (ATF) and Tangential Flow Filtration (TFF) techniques; 3) Fiber matrix based CellTank™ prototype; 4) Glass stirred tank bioreactor equipped with ATF. In all the systems, extremely high viable cell densities above 130 million viable cells per milliliter (MVC/mL) up to 214 MVC/mL were achieved. Steady states were maintained and studied at 20-30 MVC/mL and 100-130 MVC/mL for process development. Perfusion rate selection based on cell specific perfusion rate (CSPR) was systematically investigated and exometabolome study was performed to explore the metabolic footprint of HCDC perfusion process.

  • 371.
    Zhang, Ye
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Zhan, Caijuan
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Girod, Pierre-Alain
    Martiné, Alexandra
    Chotteau, Veronique
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Optimization of the cell specific perfusion rate in high cell density perfusion processManuskript (preprint) (Annet vitenskapelig)
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