The Experts below are selected from a list of 723 Experts worldwide ranked by ideXlab platform
Rodriguez S Couto - One of the best experts on this subject based on the ideXlab platform.
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decolouration of industrial azo dyes by crude laccase from Trametes Hirsuta
2007Co-Authors: Rodriguez S CoutoAbstract:The decolouration of several azo dyes, commonly used in the leather industry, by crude laccase obtained from Trametes Hirsuta cultivation was assessed. Among the six dyes studied four showed a decolouration percentage higher than 50% in 4 h, whereas the other two showed more resistance to degradation. These results show the ability of laccase towards different dye structures as well as its enormous potential for the decolouration of recalcitrant azo dyes.
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laccase from Trametes Hirsuta grown on paper cuttings application to synthetic dye decolorization at different ph values
2007Co-Authors: Rodriguez S CoutoAbstract:The potential of paper cuttings to produce laccase from Trametes Hirsuta grown under solid-state conditions was investigated. In addition, cultures were also grown on barley bran, a support commonly used in solid-state fermentation (SSF), for comparison. Paper cutting cultures showed a maximum individual laccase activity of 7695 U/L on day 9. In addition, the ability to decolorize two structurally different dyes (Indigo Carmine and Lissamine Green B) by the extracellular liquid from both paper and barley bran cultures at pH values between 2 and 11 was analyzed. Laccase-containing enzyme preparations from both cultures decolorized the dyes tested at pH values between 4 and 7 and, in addition, the laccase-containing enzyme preparation from paper cutting cultures was also able to decolorize the dyes tested at alkaline pH values. This is a very interesting and novel result, since no decolorization by fungal laccases has been reported until recently at pH values higher than pH 7.
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the effect of violuric acid on the decolourization of recalcitrant dyes by laccase from Trametes Hirsuta
2007Co-Authors: Rodriguez S Couto, M A SanromanAbstract:Abstract In this paper, the effect of the redox mediator violuric acid (VA) on the decolourization of the two recalcitrant acid dyes C.I. Acid Red 97 and C.I. Acid Green 26 by crude laccase obtained from Trametes Hirsuta cultures was investigated. The laccase–mediator system (LMS) led to a higher extent of decolourization in shorter times than that obtained without mediator addition, especially for C.I. Acid Red 97 infact the dye was almost totally decolourized (about 90%) in only 3 min. In addition, the stability of laccase against VA was also assessed.
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laccase activity from the fungus Trametes Hirsuta using an air lift bioreactor
2006Co-Authors: Rodriguez S Couto, A Rodriguez, R R M Paterson, Nelson Lima, J A TeixeiraAbstract:Aim: To produce high laccase activities from the white-rot fungus Trametes Hirsuta in an in-house air-lift bioreactor (ALB). Methods and Results: Trametes Hirsuta was grown in a 6-l ALB. A fed-batch strategy with glycerol as an addition resulted in maximum laccase activity of 19 400 U l )1 , which was the highest reported from the fungus. Conclusion: The ALB configuration with additional glycerol resulted in high laccase activities. Significance and Impact of the Study: This study provides useful information on how to produce high concentrations of laccase.
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decolourization of synthetic dyes by Trametes Hirsuta in expanded bed reactors
2006Co-Authors: Rodriguez S Couto, Emilio Rosales, M A SanromanAbstract:The present paper studies the decolourization of different synthetic dyes (Indigo Carmine, Bromophenol Blue, Methyl Orange and Poly R-478) by the white-rot fungus Trametes Hirsuta at bioreactor scale under solid-state conditions, operating with ground orange peelings as a support-substrate. Dye decolourization was performed in both batch and continuous mode. Batch cultivation led to high decolourization percentages in a short time (100% for Indigo Carmine in 3 h and 85% for Bromophenol Blue in 7 h). As for continuous cultivation, different hydraulic retention times (HRT) were studied (0.8, 1, 1.5 and 3 d). The highest decolourization percentages were obtained operating at a HRT of 3 d, especially for the dyes Methyl Orange and Poly R-478 (81.4% and 46.9%, respectively). This is a very interesting result, since there are few studies dealing with the continuous decolourization of dyes at bioreactor scale by fungal laccases.
Sergey Shleev - One of the best experts on this subject based on the ideXlab platform.
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third generation oxygen amperometric biosensor based on Trametes Hirsuta laccase covalently bound to graphite electrode
2015Co-Authors: Cristina Gutierrezsanchez, Sergey Shleev, Antonio L De Lacey, Marcos PitaAbstract:The response of low-density graphite electrodes hosting Trametes Hirsuta laccase in a direct electron transfer regime is presented for real-time analysis of O2 concentrations. The use of contrasting immobilisation methods developed for biocathodes affords good reproducibility and reliability of the amperometric biosensor, which shows a limit of detection below 1 µM and a sensitivity slightly higher than 60 nA cm−2 M−1.
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Comparison of bioelectrocatalysis at Trichaptum abietinum and Trametes Hirsuta laccase modified electrodes
2014Co-Authors: Marius Dagys, Sergey Shleev, Peter Lamberg, Gediminas Niaura, Irina Bachmatova, Liucija Marcinkeviciene, Rolandas Meškys, Juozas Kulys, Thomas Arnebrant, Tautgirdas RuzgasAbstract:Bioelectrocatalytic reduction of oxygen to water at electrodes modified with gold nanoparticles and a new laccase from Trichaptum abietinum (TaLc) was studied. The bioelectrocatalytic current was found to be much higher at TaLc modified electrodes than at similarly prepared electrodes modified with a broadly used laccase from Trametes Hirsuta (ThLc). To explain this difference the bioelectrocatalysis was described in terms of kinetic rate constants based on simultaneous cyclic voltammetry and quartz crystal microbalance measurements. From analysis of the rate constants both laccases appeared to possess similar rates (k0) of direct electron transfer. However, the enzyme turnover (kcat) was about three-fold higher for gold nanoparticle bound TaLc than for ThLc, calculated using surface concentration of enzyme established by QCM-D. Near reversible potential-induced reorientation of adsorbed proteins was observed by surface enhanced Raman spectroscopy.
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Direct electron transfer of Trametes Hirsuta laccase adsorbed at unmodified nanoporous gold electrodes
2012Co-Authors: Urszula Salaj-kosla, Sergey Shleev, Sascha Pöller, Wolfgang Schuhmann, Edmond MagnerAbstract:The enzyme Trametes Hirsuta laccase undergoes direct electron transfer at unmodified nanoporous gold electrodes, displaying a current density of 28μA/cm(2). The response indicates that ThLc was immobilised at the surface of the nanopores in a manner which promoted direct electron transfer, in contrast to the absence of a response at unmodified polycrystalline gold electrodes. The bioelectrocatalytic activity of ThLc modified nanoporous gold electrodes was strongly dependent on the presence of halide ions. Fluoride completely inhibited the enzymatic response, whereas in the presence of 150mM Cl(-), the current was reduced to 50% of the response in the absence of Cl(-). The current increased by 40% when the temperature was increased from 20°C to 37°C. The response is limited by enzymatic and/or enzyme electrode kinetics and is 30% of that observed for ThLc co-immobilised with an osmium redox polymer.
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design of a bioelectrocatalytic electrode interface for oxygen reduction in biofuel cells based on a specifically adapted os complex containing redox polymer with entrapped Trametes Hirsuta laccase
2010Co-Authors: Yvonne Ackermann, Sergey Shleev, Dmitrii A Guschin, Kathrin Eckhard, Wolfgang SchuhmannAbstract:Abstract The design of the coordination shell of an Os-complex and its integration within an electrodeposition polymer enables fast electron transfer between an electrode and a polymer entrapped high-potential laccase from the basidiomycete Trametes Hirsuta . The redox potential of the Os 3+/2+ -centre tethered to the polymer backbone (+ 720 mV vs. NHE) is perfectly matching the potential of the enzyme (+ 780 mV vs. NHE at pH 6.5). The laccase and the Os-complex modified anodic electrodeposition polymer were simultaneously precipitated on the surface of a glassy carbon electrode by means of a pH-shift to 2.5. The modified electrode was investigated with respect to biocatalytic O 2 reduction to H 2 O. The proposed modified electrode has potential applications as biofuel cell cathode.
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carbon ceramic electrodes modified with laccase from Trametes Hirsuta fabrication characterization and their use for phenolic compounds detection
2007Co-Authors: Behzad Haghighi, Sergey Shleev, Amir Rahmatipanah, Lo GortonAbstract:Fungal laccase (Lc) from the basidiomycete Trametes Hirsuta was immobilized on top of a carbon ceramic electrode using physical absorption. Direct, unmediated heterogeneous electron transfer between Lc and the carbon ceramic electrode (CCE) under aerobic conditions was shown. The bioelectrocatalytic reduction of oxygen on Lc-CCE started at about 430 mV vs. Ag vertical bar AgCl vertical bar KClsat at pH 3.5 and moved with about 57 mV in the cathodic region per pH unit. The Lc-modified CCE was then used as a biosensing detection element in a single line flow injection system for the amperometric determination of a variety of phenolic substrates of the enzyme. The experimental conditions were studied and optimized for catechol serving as a model compound. Statistical aspects were applied and the sensor characteristics and Michaelis-Menten constants of the investigated phenolic compounds were calculated and compared with those obtained for solid graphite electrodes modified with Trametes Hirsuta laccase. The results showed that the CCE based biosensor in comparison with the solid graphite based biosensor offers a lower detection limit, a wider linear dynamic range, and excellent operational stability with no sensor passivation, indicating that the sol - gel lattice improves the electrochemical behavior of the biosensor.
Dede Heri Yuli Yanto - One of the best experts on this subject based on the ideXlab platform.
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biodegradation and biodetoxification of batik dye wastewater by laccase from Trametes Hirsuta edn 082 immobilised on light expanded clay aggregate
2021Co-Authors: Dede Heri Yuli Yanto, Sita Heris Anita, Maulida Oktaviani, Oktan Dwi Nurhayat, Kharisma Panji Ramadhan, Maria Andriani Guntoro, Mokhammad Fajar Pradipta, Takashi WatanabeAbstract:The biodegradation and biodetoxification of batik industrial wastewater by laccase enzyme immobilised on light expanded clay aggregate (LECA) were investigated. Laccase from Trametes Hirsuta EDN 082 was covalently immobilised by modifying the LECA surface using (3–aminopropyl)trimethoxysilane and glutaraldehyde. The enzymatic characterisation of LECA–laccase showed promising results with an enzyme loading of 6.67 U/g and an immobilisation yield of 66.7% at the initial laccase activity of 10 U/g LECA. LECA-laccase successfully degraded batik industrial wastewater containing indigosol dye up to 98.2%. In addition, the decolorisation extent was more than 95.4% after four cycles. The phytotoxicity assessment of Vigna radiata and the microbial toxicity of two pathogenic bacteria, Bacillus subtilis and Pseudomonas aeruginosa, showed biodetoxification of treated batik dye wastewater. The characterisation using 3D light microscopy, scanning electron microscopy and Fourier transform infrared for LECA–laccase confirmed that laccase was successfully immobilised on LECA, and the decolorisation achieved through the combination of adsorption and enzymatic degradation. This study offers an environmentally friendly, effective and affordable LECA–laccase as a method for batik dye wastewater treatment.
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biodegradation and metabolic pathway of anthraquinone dyes by Trametes Hirsuta d7 immobilized in light expanded clay aggregate and cytotoxicity assessment
2021Co-Authors: Rafiqul Alam, Dede Heri Yuli Yanto, Fenny Clara Ardiati, Nissa Nurfajrin Solihat, Badrul Alam, Sanghan Lee, Takashi Watanabe, Sunghwan KimAbstract:Biodegradation and metabolic pathways of three anthraquinone dyes, Reactive Blue 4 (RB4), Remazol Brilliant Blue - R (RBBR), and Acid Blue 129 (AB129) by Trametes Hirsuta D7 fungus immobilized in light expanded clay aggregate (LECA) were investigated. Morphological characteristics observed with scanning electron microscope (SEM) showed successful immobilization of the fungus in LECA. Based on UV absorbance measurement, immobilized T. Hirsuta D7 effectively degraded 90%, 95%, and 96% of RB4, RBBR and AB129, respectively. Metabolites were identified with high-resolution mass spectrometry (HRMS) and degradation pathway of the dyes by T. Hirsuta D7 was proposed. Toxicity assay on human dermal fibroblast (HDF) showed that anthraquinone dyes exhibits significant toxicity of 35%, 40%, and 34% reduction of cell viability by RB4, RBBR, and AB129, respectively. Fungal treatment resulted in an abatement of the toxicity and cell viability was increased up to 94%. The data clearly showed the effectiveness of immobilized T. Hirsuta D7 in LECA on detoxification of anthraquinone dyes. This study provides potential and fundamental understanding of wastewater treatment using the newly isolated fungus T. Hirsuta D7.
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immobilization of laccase from Trametes Hirsuta edn 082 in light expanded clay aggregate for decolorization of remazol brilliant blue r dye
2020Co-Authors: Sita Heris Anita, Fahriya Puspita Sari, Fenny Clara Ardiati, Maulida Oktaviani, Oktan Dwi Nurhayat, Kharisma Panji Ramadhan, Dede Heri Yuli YantoAbstract:Abstract Immobilization is a method to enhance the activity, the stability and the reusability of enzymes. This study aims to evaluate the capacity of laccase from Trametes Hirsuta EDN 082 immobilized in the activated Light Expanded Clay Aggregate (LECA) for dye decolorization. The effects of enzyme loading, the dosage of immobilized laccase-LECA, and the enzyme reusability were investigated. The results showed that the immobilization yield laccase-LECA was 78.78%. One gram of immobilized laccase-LECA with 4.65 U/g of enzyme loading decolorized 99.29% Remazol Brilliant Blue R (RBBR) dye for 3 h. The immobilized laccase-LECA reusability was up to 6 number cycles with 79.67% of the remaining decolorization rate. Immobilized laccase also shifted wide pH and temperature values compare to free enzyme. The kinetic study showed that the specific decolorization rate increases with an increase in initial dye concentration up to 700 and 900 mg/L for laccase-LECA and free laccase, respectively.
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sequential production of ligninolytic xylanolytic and cellulolytic enzymes by Trametes Hirsuta aa 017 under different biomass of indonesian sorghum accessions induced cultures
2020Co-Authors: Ade Andriani, Sita Heris Anita, Dede Heri Yuli Yanto, Alika Maharani, Hartinah Pratiwi, Dwi Astuti, Isa Nuryana, Eva Agustriana, Ario Betha Juanssilfero, Urip PerwitasariAbstract:Abstract Biorefinery concept encourages agricultural sector for developing zero waste technology. Utilization of agricultural residues for producing valuable products has become an important strategy to be investigated. Sorghum (Sorghum bicolor L) biomass, as one of agricultural residues, is potentially of great significance as a lignocellulosic material for production of lignocellulolytic enzymessuch as Laccase (Lac), Manganese Peroxidase (MnP), and Lignin Peroxidase (LiP), Cellulases (Cel), and Xylanases (Xyl). In the present study, 13 Indonesian sorghum accessions (with different lignocellulosic characteristics) were investigated due to their potency as potential substrate for production ligninolytic, hemicellulolytic, and cellulolytic enzymes by Trametes Hirsuta AA-017 which has ability to sequentially produce the enzymes. Three accessions (Samurai, 4183, Kawali) were found as most suitable materials for enzymes production by the fungus with highest enhancement were more than 8000 and 71-fold, respectively for Lac and MnP. Total lignin in the biomass has shown a positive correlation with Lac (Pearson coefficient = 0.630; P
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Decolorization of Synthetic Dyes by Ligninolytic Enzymes from Trametes Hirsuta D7
2019Co-Authors: Sita Heris Anita, Fahriya Puspita Sari, Dede Heri Yuli YantoAbstract:The ability of ligninolytic enzymes from Trametes Hirsuta D7 to decolorize several synthetic dyes was investigated. A crude enzyme powder was produced by fermenting oil palm empty fruit bunch fibers for one month. The dye decolorization by the enzymes occurred at an efficient 0.25 U/mL. The enzymes degraded 100, 200, 300, 400, and 500 ppm Remazol Brilliant Blue R (RBBR) within 7 h by 95.57 ± 0.32%, 93.46 ± 3.09%, 91.84 ± 0.65%, 86.44 ± 0.97%, and 82.14 ± 0.52%, respectively. The enzyme also decolorized anthraquinone (Acid Blue 129), monoazo (Acid Orange 7), diazo (Reactive Black 5), and trimethyl methane (Methyl Violet) dyes within 7 h by 94.59 ± 7.97%, 13.99 ± 0.30%, 7.61 ± 0.01%, and 7.59 ± 0.18%, respectively. Addition of MnSO4, H2O2, and violuric acid enhanced the dye decolorization rate by up to 10-fold. This study shows the potential for application of ligninolytic enzymes from T. Hirsuta D7 in the treatment of wastewater effluent of textile industries
Lo Gorton - One of the best experts on this subject based on the ideXlab platform.
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carbon ceramic electrodes modified with laccase from Trametes Hirsuta fabrication characterization and their use for phenolic compounds detection
2007Co-Authors: Behzad Haghighi, Sergey Shleev, Amir Rahmatipanah, Lo GortonAbstract:Fungal laccase (Lc) from the basidiomycete Trametes Hirsuta was immobilized on top of a carbon ceramic electrode using physical absorption. Direct, unmediated heterogeneous electron transfer between Lc and the carbon ceramic electrode (CCE) under aerobic conditions was shown. The bioelectrocatalytic reduction of oxygen on Lc-CCE started at about 430 mV vs. Ag vertical bar AgCl vertical bar KClsat at pH 3.5 and moved with about 57 mV in the cathodic region per pH unit. The Lc-modified CCE was then used as a biosensing detection element in a single line flow injection system for the amperometric determination of a variety of phenolic substrates of the enzyme. The experimental conditions were studied and optimized for catechol serving as a model compound. Statistical aspects were applied and the sensor characteristics and Michaelis-Menten constants of the investigated phenolic compounds were calculated and compared with those obtained for solid graphite electrodes modified with Trametes Hirsuta laccase. The results showed that the CCE based biosensor in comparison with the solid graphite based biosensor offers a lower detection limit, a wider linear dynamic range, and excellent operational stability with no sensor passivation, indicating that the sol - gel lattice improves the electrochemical behavior of the biosensor.
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Direct heterogeneous electron transfer reactions of Trametes Hirsuta Laccase at bare and thiol-modified gold electrodes
2006Co-Authors: Shleev Sergey, Ruzgas Tautgirdas, Pita Marcos, Yaropolov Alexander, Lo GortonAbstract:Direct electron transfer reactions of high redox potential Trametes Hirsuta laccase on bare and 15 different thiol-modified gold electrodes were studied using cyclic voltammetry and potentiometry: Well-pronounced Faradaic processes were obtained for the enzyme adsorbed on bare gold, whereas reproducible and stable electrochemistry was obtained only when 4-aminothiophenol was used for gold modification. Moreover, the laccase-4-aminothiophenol-modified gold electrode showed the highest value of the steady-state potential under aerobic conditions equal to 660 mV vs. NHE compared with the other 15 different thiol modified electrodes and also the bare electrodes with immobilized enzyme. However, this value is still too far away from the equilibrium potential of the oxygen electrode and Trametes Hirsuta laccase-modified graphite electrode, for which a well-pronounced high potential process of oxygen bioelectroreduction is shown at 800 mV vs. NHE. Possible mechanisms of the enzyme function on bare and thiol-modified gold electrodes are discussed in correlation with the structure and orientation of the enzyme on the surface of the various electrodes
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Direct Heterogeneous Electron Transfer Reactions of Trametes Hirsuta Laccase at Bare and Thiol‐Modified Gold Electrodes
2006Co-Authors: Shleev Sergey, Ruzgas Tautgirdas, Pita Marcos, Yaropolov Alexander i., Lo GortonAbstract:Direct electron transfer reactions of high redox potential Trametes Hirsuta laccase on bare and 15 different thiol‐modified gold electrodes were studied using cyclic voltammetry and potentiometry. Well‐pronounced Faradaic processes were obtained for the enzyme adsorbed on bare gold, whereas reproducible and stable electrochemistry was obtained only when 4‐aminothiophenol was used for gold modification. Moreover, the laccase‐4‐aminothiophenol‐modified gold electrode showed the highest value of the steady‐state potential under aerobic conditions equal to 660 mV vs. NHE compared with the other 15 different thiol modified electrodes and also the bare electrodes with immobilized enzyme. However, this value is still too far away from the equilibrium potential of the oxygen electrode and Trametes Hirsuta laccase‐modified graphite electrode, for which a well‐pronounced high potential process of oxygen bioelectroreduction is shown at 800 mV vs. NHE. Possible mechanisms of the enzyme function on bare and thiol‐modified gold electrodes are discussed in correlation with the structure and orientation of the enzyme on the surface of the various electrodes.The work was supported by the Swedish Research Council, the Swedish Institute (SI), the Consejería de Educación de la Comunidad de Madrid, and the European Social Funding (F. S. E.). The SI is acknowledged for the support of a postdoctoral fellowship for S. S.Peer reviewe
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Characterization of graphite electrodes modified with laccases from Trametes Hirsuta and Cerrena unicolor and their use for flow injection amperometric determination of some phenolic compounds
2005Co-Authors: Behzad Haghighi, Lo Gorton, Anna Jarosz-wilkołazka, Tautgirdas Ruzgas, Andrzej LeonowiczAbstract:Fungal laccases from Trametes Hirsuta and Cerrena unicolor were immobilized on spectroscopic graphite electrodes using physical absorption. The laccase-modified graphite electrodes were mounted as the working electrodes into a wall jet flow through an electrochemical cell and then used for flow injection amperometric determination of different phenolic compounds of relevance both for environmental and clinical analysis. At optimum conditions for determination of catechol serving as a model compound, the output response signals of the laccase-modified electrodes were recorded as the result of injections of 50 μL of solutions with different phenolic substrates into the carrier citrate buffer solution with a working potential of −50 mV vs. Ag|AgCl. Statistical aspects were applied and the Michaelis–Menten constants evaluated were correlated to the chemical structure of the investigated phenolic compounds.
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Characterization of graphite electrodes modified with laccases from Trametes Hirsuta and Cerrena unicolor and their use for flow injection amperometric determination of some phenolic compounds
2005Co-Authors: Haghighi Behzad, Lo Gorton, Jarosz-wilkolazka A, Ruzgas Tautgirdas, Leonowicz AAbstract:Fungal laccases from Trametes Hirsuta and Cerrena unicolor were immobilized on spectroscopic graphite electrodes using physical absorption. The laccase-modified graphite electrodes were mounted as the working electrodes into a wall jet flow through an electrochemical cell and then used for flow injection amperometric determination of different phenolic compounds of relevance both for environmental and clinical analysis. At optimum conditions for determination of catechol serving as a model compound, the output response signals of the laccase-modified electrodes were recorded as the result of injections of 50 mu L of solutions with different phenolic Substrates into the carrier citrate buffer solution with a working potential of -50mV vs. Ag vertical bar AgCl. Statistical aspects were applied and the Michaelis-Menten constants evaluated were correlated to the chemical structure or the investigated phenolic compounds
Daria V Vasina - One of the best experts on this subject based on the ideXlab platform.
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relation between lignin molecular profile and fungal exo proteome during kraft lignin modification by Trametes Hirsuta le bin 072
2021Co-Authors: Konstantin V Moiseenko, O A Glazunova, Daria V Vasina, Olga S Savinova, N A Kulikova, Alexander Zherebker, Evgeny N Nikolaev, Tatiana V. FedorovaAbstract:Abstract The process of kraft lignin modification by the white-rot fungus Trametes Hirsuta was investigated using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI FT-ICR MS), and groups of systematically changing compounds were delineated. In the course of cultivation, fungus tended to degrade progressively more reduced compounds and produced more oxidized ones. However, this process was not gradual – the substantial discontinuity was observed between 6th and 10th days of cultivation. Simultaneously, the secretion of ligninolytic peroxidases by the fungus was changing in a cascade manner – new isoenzymes were added to the mixture of the already secreted ones, and once new isoenzyme appeared both its relative quantity and number of isoforms increased as cultivation proceeded. It was proposed, that the later secreted peroxidases (MnP7 and MnP1) possess higher substrate affinity for some phenolic compounds and act in more specialized manner than the early secreted ones (MnP5 and VP2).
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the minor recombinant laccase isozymes of Trametes Hirsuta 072 preparation and properties
2019Co-Authors: Olga S Savinova, Daria V Vasina, I N Zorov, A P Sinitsyn, T V FedorovaAbstract:Laccases (EC 1.10.3.2) are multicopper polyphenol oxidases found in various organisms, in particular, in fungi. Fungal laccases are encoded by multigene families, which can contain up to 17 genes. However, not all isozymes can be obtained from native producers. Previous studies have shown that the filamentous fungus Penicillium canescens is a promising object for the heterologous expression of various laccase isozymes of Trametes Hirsuta 072. In this work, the cultivation conditions of P. canescens strains, recombinant producers of T. Hirsuta heterologous minor laccase, are optimized. The optimization of the rLacD and rLacF purification method increased their yield and specific activity of rLacD. In addition, the melting points of minor laccase isoenzymes are measured and the glycosylation of isoenzymes is studied.
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evolutionary relationships between the laccase genes of polyporales orthology based classification of laccase isozymes and functional insight from Trametes Hirsuta
2019Co-Authors: Olga S Savinova, Tatiana V. Fedorova, Konstantin V Moiseenko, T V Tyazhelova, E A Vavilova, A M Chulkin, Daria V VasinaAbstract:Laccase is one of the oldest known and intensively studied fungal enzymes capable of oxidizing recalcitrant lignin-resembling phenolic compounds. It is currently well established that fungal genomes almost always contain several non-allelic copies of laccase genes (laccase multigene families); nevertheless, many aspects of laccase multigenicity, for example, their precise biological functions or evolutionary relationships, are mostly unknown. Here, we present a detailed evolutionary analysis of the sensu stricto laccase genes (CAZy – AA1_1) from fungi of the Polyporales order. The conducted analysis provides a better understanding of the Polyporales laccase multigenicity and allows for the systemization of the individual features of different laccase isozymes. In addition, we provide a comparison of the biochemical and catalytic properties of the four laccase isozymes from Trametes Hirsuta and suggest their functional diversification within the multigene family.
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properties of two laccases from the Trametes Hirsuta 072 multigene family twins with different faces
2017Co-Authors: Olga S Savinova, Konstantin V Moiseenko, T V Tyazhelova, E A Vavilova, Daria V VasinaAbstract:Abstract Utilization of laccases in biotechnology and bioremediation has created a strong demand for the characterization of new enzymes and an increase in production of known laccases. Thus, additional research into these enzymes is critically needed. In this study, we report a comparative study of the biochemical and transcriptional properties of two different laccase isozymes from Trametes Hirsuta 072 – the constitutive and inducible forms. A recombinant LacC enzyme was expressed in Penicillium canescens to characterize its properties. LacC is single-purpose enzyme, unlike LacA, which can operate efficiently under a wide range of temperatures and pHs (55–70 °C and pH 3–5, respectively). LacC has a lower RedOx potential than LacA and does not oxidize substrates containing amine groups. Expression of the lacC gene was selective compared to that of the lacA gene and increased significantly in the presence of complex synthetic compounds such as dyes and xenobiotics. This study shows that laccases from the multigene families of basidiomycetes differ significantly in their properties, thus providing a complementary effect during lignin degradation.
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lignin degrading peroxidases in white rot fungus Trametes Hirsuta 072 absolute expression quantification of full multigene family
2017Co-Authors: Daria V Vasina, Tatiana V. Fedorova, Konstantin V Moiseenko, T V TyazhelovaAbstract:Ligninolytic heme peroxidases comprise an extensive family of enzymes, which production is characteristic for white-rot Basidiomycota. The majority of fungal heme peroxidases are encoded by multigene families that differentially express closely related proteins. Currently, there were very few attempts to characterize the complete multigene family of heme peroxidases in a single fungus. Here we are focusing on identification and characterization of peroxidase genes, which are transcribed and secreted by basidiomycete Trametes Hirsuta 072, an efficient lignin degrader. The T. Hirsuta genome contains 18 ligninolytic peroxidase genes encoding 9 putative lignin peroxidases (LiP), 7 putative short manganese peroxidases (MnP) and 2 putative versatile peroxidases (VP). Using ddPCR method we have quantified the absolute expression of the 18 peroxidase genes under different culture conditions and on different growth stages of basidiomycete. It was shown that only two genes (one MnP and one VP) were prevalently expressed as well as secreted into cultural broth under all conditions investigated. However their transcriptome and protein profiles differed in time depending on the effector used. The expression of other peroxidase genes revealed a significant variability, so one can propose the specific roles of these enzymes in fungal development and lifestyle.