RESUMEN
In this study, antimicrobial pigment cinnabarinic acid (CA) was produced from Pycnoporus cinnabarinus in laboratory-scale batch cultures. Magnetic poly(ethylene glycol dimethacrylate-N-methacryloyl-l-tryptophan methyl ester) [m-poly(EGDMA-MATrp)] beads (average diameter = 53-103 µm) were synthesized by copolymerizing of N-methacryloyl-l-tryptophan methyl ester (MATrp) with ethylene glycol dimethacrylate (EGDMA) in the presence of magnetite (Fe3O4) and used for the adsorption of CA. The m-poly(EGDMA-MATrp) beads were characterized by N2 adsorption/desorption isotherms (Brunauer Emmet Teller), X-ray photoelecron spectroscopy, scanning electron microscopy, infrared spectroscopy, thermal gravimetric analysis, electron spin resonance and swelling studies. The efficiency of m-poly(EGDMA-MATrp) beads for separation of CA from culture fluid was evaluated. The effects of pH, initial concentration, contact time and temperature on adsorption were analyzed. The maximum CA adsorption capacity of the m-poly(EGDMA-MATrp) beads was 272.9 mg g(-1) at pH 7.0, 25 °C. All the isotherm data can be fitted with the Langmuir, Freundlich and Dubinin-Radushkevich isotherm models. The adsorption process obeyed pseudo-second-order kinetic model. Thermodynamic parameters ΔH = 5.056 kJ mol(-1), ΔS = 52.44 J K(-1) mol(-1) and ΔG = -9.424 kJ mol-(1) to -11.27 kJ mol-(1) with the rise in temperature from 4 to 40 °C indicated that the adsorption process was endothermic and spontaneous.
Asunto(s)
Imanes , Microesferas , Oxazinas/química , Oxazinas/aislamiento & purificación , Pycnoporus/metabolismo , Adsorción , Reactores Biológicos , Concentración de Iones de Hidrógeno , Metacrilatos , Polietilenglicoles , TermodinámicaRESUMEN
Poly(3-hydroxybutyrate) is nontoxic and biodegradable, with good biocompatibility and potential support for long-term implants. For this reason, it is a good support for enzyme immobilization. Enzyme immobilization could not be done directly because poly(3-hydroxybutyrate) has no functional groups. Therefore, modification should be done for enzyme immobilization. In this study, methacrylic acid was graft polymerized to poly(3-hydroxybutyrate) and thrombin was immobilized to polymethacrylic acid grafted poly(3-hydroxybutyrate). In fact, graft polymerization of methacrylic acid to poly(3-hydroxybutyrate) and thrombin immobilization was a model study. Biomolecule immobilized poly(3-hydroxybutyrate) could be used as an implant. Thrombin was selected as a biomolecule for this model study and it was immobilized to methacrylic acid grafted poly(3-hydroxybutyrate). Then the developed product was used to stop bleeding.
Asunto(s)
Enzimas Inmovilizadas/química , Hidroxibutiratos/química , Poliésteres/química , Ácidos Polimetacrílicos/química , Trombina/química , Coagulación Sanguínea/efectos de los fármacos , Pruebas de Enzimas , Enzimas Inmovilizadas/farmacología , Humanos , Cinética , Modelos Químicos , Espectroscopía Infrarroja por Transformada de Fourier , Temperatura , Trombina/farmacología , Tiempo de Coagulación de la Sangre TotalRESUMEN
The aim of this work was to determine new, different and low-cost substrates that can be used for enzyme production from the white rot fungus Trametes versicolor (ATCC 11235) by taking advantage of the broad substrate specificity of pyranose 2-oxidase. In this report, we investigated the production of pyranose 2-oxidase from T. versicolor (ATCC 11235) using ten different agricultural residues such as clover straw, almond shells, hazelnut cobs, grass and others. Pyranose 2-oxidase activity was determined as 2.332 U/g at the 9th day in a submerged culture containing clover straw and tap water shaken at 150 rpm and 26°C, and the optimum clover straw concentration was determined to be 12 g/l. The effects of different glucose, nitrogen and phosphate sources on the production of pyranose 2-oxidase were studied in the clover straw medium. Analyses of biomass, protein, reduced sugar and nitrogen concentrations were also monitored in a clover straw medium that did not contain carbon or nitrogen and phosphate sources under the parameters determined. The produced pyranose 2-oxidase was used for improving the properties of cotton fabrics.
Asunto(s)
Deshidrogenasas de Carbohidratos/metabolismo , Gossypium/metabolismo , Lignina/metabolismo , Trametes/enzimología , Medios de Cultivo/química , Glucosa/metabolismo , Nitrógeno/metabolismo , Fosfatos/metabolismo , Temperatura , Factores de Tiempo , Trametes/crecimiento & desarrolloRESUMEN
The biodegradation kinetics of o-cresol was examined by acclimatized P. putida DSM 548 (pJP4) in batch experiments at varying initial o-cresol concentrations (from 50 to 500 mg/L). The kinetic parameters of o-cresol aerobic biodegradation were estimated by using the Haldane substrate inhibition equation. The biodegradation kinetics of o-cresol was investigated. In batch culture reactors, the Maximum specific growth rate (μmax), Monod constant (Ks) and the inhibition constant (Ki) were established as 0.519 h-1, 223.84 mg/L and 130.883 mg/L, respectively. o-cresol biodegradation in a batch-recirculation bioreactor system by immobilized P. putida was also studied. The recycled packed bed reactor system, which was composed of Ca-alginate beads and pumice on which cells immobilized, has been performed to determine possible stability for further developments.
Asunto(s)
Biodegradación Ambiental , Cresoles/metabolismo , Pseudomonas putida/química , Reactores Biológicos , Células Inmovilizadas , Fenoles/metabolismo , CinéticaRESUMEN
In this study, lyophilized Trametes versicolor biomass is used as a sorbent for biosorption of a textile dye, Sirius Blue K-CFN, from an aqueous solution. The batch sorption was studied with respect to dye concentration, adsorbent dose and equilibrium time. The effect of pH and temperature on dye uptake was also investigated and kinetic parameters were determined. Optimal initial pH (3.0), equilibrium time (2 hrs), initial dye concentration ( 100 mg l-1) and biomass concentration (1.2 mg l-1) were determined at 26ºC. The maximum biosorption capacity (q max) of Sirius Blue K-CFN dye on lyophilized T. versicolor biomass is 62.62 mg/g. The kinetic and isotherm studies indicated that the biosorption process obeys to a pseudo-second order model and Langmuir isotherm model. In addition, the biosorption capacities of fungal biomass compared to other well known adsorbents such as activated carbon and Amberlite, fungal biomass biosorptions capacities were found to be more efficient.
Asunto(s)
Trametes/química , Biomasa , Basidiomycota/química , Colorantes , Compuestos Azo/química , Liofilización , Concentración de Iones de Hidrógeno , Isoterma , Cinética , TemperaturaRESUMEN
Depending upon their structure, azo- and anthraquinonic dyes are the two major classes and together represent 90% of all organic colorants. Adsorption of dye molecules onto a sorbent can be an effective, low-cost method of color removal. Most of the techniques used for removal of dyes are of high production cost, and the regeneration also makes them uneconomical. There is much interest in the development of cheaper and effective newer materials for use as adsorbents. Molecular imprinting is a new kind of materials that can be alternative adsorbents. In this study, molecularly imprinted polymers of three textile dyes (Cibacron Orange P-4R, Cibacron Red P-4B, Cibacron Black PSG) were prepared. Methacrylic acid was used as a monomer for red and orange dyes and acrylamide was used for black dye. Methanol:acetonitrile was used as a porogen. The selective recognition ability of the molecularly imprinted polymers was studied by an equilibrium-adsorption batch method. The adsorption data are for Cibacron Black PSG 65% and nonimprinted polymer (NIP) 25%; Cibacron Red P-4B 72% and NIP 18%; and Cibacron Orange P-4R 45% and NIP 10%, respectively. Dye-imprinted polymers were used as a solid-phase extraction material for selective adsorption from wastewater of textile factory.
Asunto(s)
Acrilamida/química , Colorantes/aislamiento & purificación , Metacrilatos/química , Impresión Molecular , Extracción en Fase Sólida/métodos , Adsorción , Polímeros/químicaRESUMEN
To investigate biodegradability by Trametes versicolor, five structurally different direct azo-dyes--Direct Black 38, Direct Blue 15 (DB 15), Direct Orange 26, Direct Green 6, and Direct Yellow 12--were studied. The DB 15 was determined as the best biodegradable dye by this white-rot fungus. Laccase and manganese peroxidase activities were monitored with the biodegradation process; it was observed that laccase played an important role in the dye degradation, while manganese peroxidase activity could not be detected. Possible degradation products also were examined by gas chromatography-mass spectrometry, but no metabolite was detected after the degradation and/or decolorization process. To enhance performance of the fungi during the degradation, Trametes versicolor cells were immobilized in alginate beads. Then, DB 15 decolorization by immobilized Trametes versicolor was studied in a small-scale packed-bed reactor. The color removal efficiency in repeated batches was found to be 98 and 93% for 50 mg/L DB 15.
Asunto(s)
Compuestos Azo/metabolismo , Biodegradación Ambiental , Trametes/metabolismo , Cromatografía de Gases y Espectrometría de Masas , Lacasa/metabolismo , Peroxidasas/metabolismo , Trametes/enzimologíaRESUMEN
Amperometric biosensors using laccase from Trametes versicolor as a bioelement were developed for 2,4-dichloro phenoxy acetic acid (2,4-D). Laccase enzyme was immobilized by gelatin and glutaraldehyde onto a Clark oxygen probe and screen printed electrodes (SPEs). Amperometric and chronoamperometric measurements were carried out with the biosensors. First, the effect of laccase activity on the biosensor performances was investigated for both biosensors, and then optimum pH and temperature and also thermal stability of the biosensors were tested. In addition, the detection ranges of some phenolic compounds were obtained by the help of calibration graphs of them. In repeatability studies, variation coefficients and standard deviations for both biosensors were also calculated by the studies done for this purposes. Finally, the biosensors were applied to the determination of 2,4-D in a real herbicide sample.
Asunto(s)
Técnicas Biosensibles/métodos , Lacasa/metabolismo , Trametes/enzimología , Xenobióticos/análisis , Ácido 2,4-Diclorofenoxiacético/análisis , Electroquímica , Herbicidas/química , Concentración de Iones de Hidrógeno , Modelos Lineales , Fenoles/análisis , Fenoles/química , Especificidad por Sustrato , Temperatura , TransductoresRESUMEN
This study presents new and alternative fungal strains for the production of ligninolytic enzymes which have great potential to use in industrial and biotechnological processes. Thirty autochthonous fungal strains were harvested from Bornova-Izmir in Turkiye. In the fresh fruitbody extracts laccase, manganese peroxidase and lignin peroxidase activities, which are the principal enzymes responsible for ligninocellulose degradation by Basidiomycetes, were screened. Spores of some of the basidiomycetes species such as Cortinarius sp., Trametes versicolor, Pleurotus ostreatus, Abortiporus biennis, Lyophyllum subglobisporium, Ramaria stricta, Ganoderma carnosum, Lactarius delicious ve Lepista nuda were isolated and investigated optimum cultivation conditions in submerged fermentation for high yields of ligninolytic enzyme production. In addition, isolated fungal strains were monitored on agar plates whether having the capability of decolorization of a textile dye Remazol Marine Blue.
Este estudo apresenta novas cepas de fungos produtores de enzimas ligninolíticas com potencial de aplicação em processos industriais e biotecnológicos. Trinta cepas de fungos autóctones foram obtidos em Bornova-Izmir, Turquia. Os extratos frescos dos corpos de frutificação foram submetidos à triagem de atividade de lacase, manganês peroxidase e lignina peroxidase, que são as principais enzimas de degradação de ligninocelulose pelos Basidiomycetes. Foram isolados esporos de Cortinarius sp, Tramnetes versicolor, Pleorotus ostreatus, Abortiporus biennis, Lyophyllum subglobisporium, Ramaria stricta, Ganoderma carnosum, Lactarius delicius ve Lepista desnuda, investigando-se as condições ótimas de cultivo em fermentação submersa para produção de enzimas ligninolíticas com elevado rendimento. Além disso, as cepas fúngicas isoladas foram monitoradas em placas de ágar quanto a capacidade de descoloramento do corante têxtil Remazole Marine Blue.
Asunto(s)
Colorantes/análisis , Basidiomycota/aislamiento & purificación , Celulosa/análisis , Esporas Fúngicas/aislamiento & purificación , Fermentación , Fungicidas Industriales/aislamiento & purificación , Lignina/análisis , Oxidorreductasas/análisis , Activación Enzimática , Métodos , Métodos , Industria TextilRESUMEN
This study presents new and alternative fungal strains for the production of ligninolytic enzymes which have great potential to use in industrial and biotechnological processes. Thirty autochthonous fungal strains were harvested from Bornova-Izmir in Turkiye. In the fresh fruitbody extracts laccase, manganese peroxidase and lignin peroxidase activities, which are the principal enzymes responsible for ligninocellulose degradation by Basidiomycetes, were screened. Spores of some of the basidiomycetes species such as Cortinarius sp., Trametes versicolor, Pleurotus ostreatus, Abortiporus biennis, Lyophyllum subglobisporium, Ramaria stricta, Ganoderma carnosum, Lactarius delicious ve Lepista nuda were isolated and investigated optimum cultivation conditions in submerged fermentation for high yields of ligninolytic enzyme production. In addition, isolated fungal strains were monitored on agar plates whether having the capability of decolorization of a textile dye Remazol Marine Blue.
RESUMEN
Amperometric biosensors using Pseudomonas putida cells as a bioelement were developed for 2,4-dichloro phenoxy acetic acid (2,4-D). After the adaptation process of Pseudomonas putida to 2,4-D, cells were immobilized onto the screen printed graphite electrodes (SPG) as well as Clark oxygen probe by gelatin and glutaraldehyde. Optimum pH, temperature, and stability of the biosensor were investigated. Substrate specificities for various phenolic compounds were also searched. In repeatability studies, variation coefficients and standard deviations for both type of systems were calculated; SPG and Clark electrodes were calculated and results are given as a comparison of two systems. Finally, the biosensors were applied to 2,4-D determination in a real herbicide sample.
Asunto(s)
Ácido 2,4-Diclorofenoxiacético/análisis , Ácido 2,4-Diclorofenoxiacético/farmacología , Bioensayo/instrumentación , Técnicas Biosensibles/instrumentación , Electroquímica/instrumentación , Pseudomonas putida/efectos de los fármacos , Pseudomonas putida/fisiología , Bioensayo/métodos , Técnicas Biosensibles/métodos , Diseño de Equipo , Análisis de Falla de Equipo , Pseudomonas putida/citología , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Optimization of pyranose-2-oxidase (P2O) production conditions from Trametes versicolor was carried out in shaking cultures containing glucose, malt, and yeast extracts; the optimum concentration values were found to be 1.5% glucose, 1.0% yeast extract, and 1.0% malt extract, pH 5.0, temperature, 26 degrees C, and agitation rate 150 rpm. For the first time, P2O production was also carried out in a stirred tank reactor (STR) with 2.2 L working volume in the optimized medium composition, and biomass, P2O activity, protein, nitrogen and glucose concentrations were also monitored besides pH and dissolved oxygen (DO). In the STR, P2O activity peaked on day 9. Partial enzyme characterization occurred and optimum pH and temperature were detected as 7.0 and 37 degrees C, respectively. K(m) value was found to be 1.009 mM.
Asunto(s)
Reactores Biológicos/microbiología , Deshidrogenasas de Carbohidratos/química , Deshidrogenasas de Carbohidratos/metabolismo , Trametes/clasificación , Trametes/enzimología , Activación Enzimática , Estabilidad de Enzimas , Especificidad de la EspecieRESUMEN
Manganese-dependent peroxidase (MnP) production was performed in an immobilized cell bioreactor in which Phanerochaete chrysosporium BKM-F-1767 was immobilized on polystyrene foam. The immobilized cell culture yielded significantly greater MnP activity than the conventional stationary liquid culture. Cultivation was carried out in batch mode; the effect of glucose concentration was investigated and growth kinetics parameters were found as, micromax=0.59 day(-1), Ks=0.33 g/L and Kss=14.5. Batch operation led to maximum MnP (770.82 U/L) in the culture medium containing 0.05% Tween 80, 10 g/L glucose, and 174 microM Mn2+ at 37 degrees C and pH 4.5. Enzyme productivity was obtained as 110.12 U/day/L.
Asunto(s)
Reactores Biológicos , Peroxidasas/biosíntesis , Phanerochaete/enzimología , Células Cultivadas , Células Inmovilizadas , Glucosa/metabolismo , Concentración de Iones de Hidrógeno , Cinética , Phanerochaete/citología , Poliestirenos/química , TemperaturaRESUMEN
Production of manganese-dependent peroxidase (MnP) by the white-rot fungus Phanerochaete chrysosporium BKM-F-1767 (ATCC 24725) was monitored during growth in different media and growth conditions. The effect of some activators of MnP production, Mn2+, Tween 80, phenylmethylsulphonylfloride (PMSF), oxygen, temperature, pH, glycerol and nitrogen was studied. Supplementing the cultures with Tween 80 (0.05 percent, v/v) and Mn2+ (174 µM) resulted a maximum MnP activity of 356 U/L which was approximately two times higher than that obtained in the control culture (without Tween 80). Decolourisation of Direct Blue 15 and Direct Green 6 (50 mg/L) was also achieved with MnP.
RESUMEN
Two biosensors based on Trametes versicolor laccase (TvL) were developed for the determination of phenolic compounds. Commercial oxygen electrode and ferrocene-modified screen-printed graphite electrodes were used for preparation of laccase biosensors. The systems were calibrated for three phenolic acids. Linearity was obtained in the concentration range 0.1-1.0muM caffeic acid, 0.05-0.2muM ferulic acid, 2.0-14.0muM syringic acid for laccase immobilised on a commercial oxygen electrode and 2.0-30.0muM caffeic acid, 2.0-10.0muM ferulic acid, 4.0-30.0muM syringic acid for laccase immobilised on ferrocene-modified screen-printed electrodes. Furthermore, optimal pH, temperature and thermal stability studies were performed with the commercial oxygen electrode. Both electrodes were used for determination of a class of phenolic acids, achieving a cheap and fast tool and an easy to be used procedure for screening real samples of human plasma.
RESUMEN
Two different flexible osmium redox polymers; poly(1-vinylimidazole)12-[Os-(4,4'-dimethyl-2,2'-di'pyridyl)2Cl2](2+/+) (osmium redox polymer I) and poly(vinylpyridine)-[Os-(N,N'-methylated-2,2'-biimidazole)3](2+/3+) (osmium redox polymer II) were investigated for their ability to efficiently "wire" Pseudomonas putida ATCC 126633 and Pseudomonas fluorescens (P. putida DSM 6521), which are well-known phenol degrading organisms, when entrapped onto cysteamine modified gold electrodes. The two Os-polymers differ in redox potential and the length of the side chains, where the Os(2+/3+)-functionalities are located. The bacterial cells were adapted to grow in the presence of phenol as the sole source of organic carbon. The performance of the redox polymers as mediators was investigated for making microbial sensors. The analytical characteristics of the microbial sensors were evaluated for determination of catechol, phenol and glucose as substrates in both batch analysis and flow analysis mode.
Asunto(s)
2,2'-Dipiridil/análogos & derivados , Técnicas Biosensibles/métodos , Imidazoles/química , Técnicas Microbiológicas/métodos , Compuestos Organometálicos/química , Polivinilos/química , Pseudomonas fluorescens/aislamiento & purificación , Pseudomonas putida/aislamiento & purificación , 2,2'-Dipiridil/química , Técnicas Biosensibles/instrumentación , Carbono/química , Catecoles/análisis , Catecoles/metabolismo , Cisteamina/química , Electroquímica , Electrodos , Glucosa/análisis , Glucosa/metabolismo , Oro/química , Concentración de Iones de Hidrógeno , Técnicas Microbiológicas/instrumentación , Compuestos Orgánicos/química , Oxidación-Reducción , Fenoles/análisis , Fenoles/metabolismo , Pseudomonas fluorescens/crecimiento & desarrollo , Pseudomonas putida/crecimiento & desarrollo , Factores de TiempoRESUMEN
An enzyme electrode suitable for paracetamol detection was developed by immobilizing laccase on a dissolved-oxygen probe surface. The immobilization procedure was achieved by means of gelatin, which was then cross-linked with glutaraldehyde. The measurement was based on the detection of oxygen consumption in relation to analyte oxidation. The optimum experimental conditions for the biosensor were investigated and the system was calibrated for paracetamol. Also the effects of three different mediators, namely HBT (1-hydroxybenzotriazole), VLA [violuric acid (5-isonitrosobarbituric acid)] and TEMPO (2,2',6,6'-tetramethylpiperidine-N-oxyl radical) were tested for the biosensor's response. As a result, it was observed that HBT has a remarkable effect on the signal by providing more oxygen consumption during the enzymatic reaction. A linear relationship between sensor responses and analyte concentrations was obtained over the concentration range 2.0-15.0 microM, whereas, in the presence of the mediator HBT, this range became 0.5-3.0 microM.
Asunto(s)
Acetaminofén/análisis , Analgésicos no Narcóticos/análisis , Técnicas Biosensibles/instrumentación , Diseño de Equipo/instrumentación , Proteínas Fúngicas/química , Lacasa/química , Oxidación-Reducción , Especificidad por SustratoRESUMEN
The modification of a graphite-epoxy composite electrode (GECE) with bacterial cells along with an analytical application are presented. Pseudomonas putida DSM 50026 was used as a biological component and the measurement was based on the respiratory activity of the cells. The optimization of working conditions of resulting biosensor (including pH and temperature) was conducted and the limit of detection was calculated as 7 microM phenol based on the signal to noise ratio. Then the system was applied for xenobiotic detection. Resulting sample signals were found to be very similar with the standard solutions having the same concentration while the recoveries of the spiked samples were close to 100%.
Asunto(s)
Técnicas Biosensibles/métodos , Compuestos Epoxi/química , Grafito/química , Pseudomonas putida/química , Técnicas Biosensibles/instrumentación , Electroquímica , Electrodos , Concentración de Iones de Hidrógeno , Fenoles/análisis , Fenoles/química , Pseudomonas putida/metabolismo , Propiedades de Superficie , Temperatura , Xenobióticos/análisisRESUMEN
A biosensor was developed by immobilizing laccase onto mercury thin film electrode (MTFE) by means of gelatin that is then crosslinked with glutaraldehyde. Mercury thin film (MTF) was deposited onto glassy carbon electrode (GCE) and the obtained biosensor was utilized for the determination of phenolic compounds. The measurement was based on the amperometric detection of oxygen consumption in relation to analyte oxidation. The optimum experimental conditions for the biosensor were investigated and the system was calibrated for both catechol and phenol. A linear relationship between sensor responses and analyte concentrations was obtained in concentration range between 0.5 x 10(-6)-5.0 x 10(-6)M for catechol and 2.5 x 10(-6)-2.0 x 10(-6)M for phenol, respectively. Mercury thin film was also formed onto the surface of screen printed graphite electrodes and applied for the catechol detection. The linearity was observed in concentration range between 2.5 x 10(-6)-3.0 x 10(-5)M.
Asunto(s)
Técnicas Biosensibles/métodos , Catecoles/análisis , Enzimas Inmovilizadas , Lacasa , Fenoles/análisis , Técnicas Biosensibles/normas , Calibración , Electrodos , Mercurio , Oxígeno/metabolismoRESUMEN
Phanerochaete chrysosporium BKMF-1767 cells were immobilized on different carriers. The optimum carrier according to adsorbed P. chrysosporium cells number (86.38%) was determined to be polystyrene foam, a novel carrier. The conditions for the immobilization of cells on polystyrene foam were optimized and determined as 50 rpm, 37 degrees C, and 2h. The results show that the adsorption of P. chrysosporium on polystyrene foam follows the Langmuir adsorption isotherm. High manganese peroxidase activity (421 U/L) and dry mass (4.7 g/L) were recovered from the batch mode polystyrene foam solid state fermentation system.