To the Mechanism of Horseradish Peroxidase-Mediated Degradation of a Recalcitrant Dye Remazol Brilliant Blue R

2001 ◽  
Vol 66 (4) ◽  
pp. 663-675 ◽  
Author(s):  
Markéta Mikšanová ◽  
Jiří Hudeček ◽  
Jan Páca ◽  
Marie Stiborová
Keyword(s):  
Molecular Weight ◽  
Horseradish Peroxidase ◽  
Oxidation Products ◽  
Brilliant Blue ◽  
Radical Mechanism ◽  
Remazol Brilliant Blue R ◽  
Radical Trapping ◽  
Optimum Ph ◽  
Mediated Oxidation

Thein vitroenzymatic metabolism of a recalcitrant dye Remazol Brilliant Blue R (RBBR) was investigated using horseradish peroxidase (HRP). At optimum pH (4.5), the apparent Michaelis constant (KM) value for the oxidation of RBBR catalyzed by HRP is 14.8 μmol l-1. HRP-mediated conversion of RBBR proceedsviaa conventional peroxidase reaction, by a sequential one-electron oxidation of two molecules of RBBR by the peroxidase Compounds I and II. The oxidation is inhibited by radical trapping agents (nicotinamide adenine dinucleotide reduced (NADH), ascorbate, glutathione). This confirms that the peroxidase-mediated oxidation of RBBR proceedsviaradical mechanism. Gel permeation profile of the RBBR oxidation products shows that the pattern of molecular weight distribution was shifted to the higher molecular weight region indicating formation of RBBR oligomers. In addition to HRP, the RBBR dye is also oxidized by another peroxidase, the mammalian lactoperoxidase.

scholarly journals Decolourisation Capabilities of Ligninolytic Enzymes Produced byMarasmius cladophyllusUMAS MS8 on Remazol Brilliant Blue R and Other Azo Dyes

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Ngieng Ngui Sing ◽  
Ahmad Husaini ◽  
Azham Zulkharnain ◽  
Hairul Azman Roslan
Keyword(s):  
Azo Dyes ◽  
Culture Medium ◽  
Laccase Activity ◽  
Dye Degradation ◽  
Ligninolytic Enzymes ◽  
Brilliant Blue ◽  
Remazol Brilliant Blue R ◽  
Orange G ◽  
Dye Decolourisation

Marasmius cladophylluswas examined for its ability to degradatively decolourise the recalcitrant dye Remazol Brilliant Blue R (RBBR) and screened for the production of ligninolytic enzymes using specific substrates. Monitoring dye decolourisation by the decrease in absorbance ratio ofA592/A500shows that the decolourisation of RBBR dye was associated with the dye degradation.Marasmius cladophyllusproduces laccase and lignin peroxidase in glucose minimal liquid medium containing RBBR. Both enzyme activities were increased, with laccase activity recorded 70 times higher reaching up to 390 U L−1on day 12. Further in vitro RBBR dye decolourisation using the culture medium shows that laccase activity was correlated with the dye decolourisation. Fresh RBBR dye continuously supplemented into the decolourised culture medium was further decolourised much faster in the subsequent round of the RBBR dye decolourisation. In vitro dye decolourisation using the crude laccase not only decolourised 76% of RBBR dye in just 19 hours but also decolourised 54% of Orange G and 33% of Congo red at the same period of time without the use of any exogenous mediator. This rapid dye decolourisation ability of the enzymes produced byM. cladophyllusthus suggested its possible application in the bioremediation of dye containing wastewater.

Transport of Low Molecular Weight Proteins through Aorta and Vena Cava of Rats and Rabbits

1972 ◽  
Vol 28 (01) ◽  
pp. 144-151 ◽  
Author(s):  
J Pinkas ◽  
Th. H Spaet
Keyword(s):  
Molecular Weight ◽  
Cytochrome C ◽  
Horseradish Peroxidase ◽  
Aortic Wall ◽  
Vena Cava ◽  
Surrounding Tissue ◽  
Molecular Weights ◽  
The Media ◽  
Horseradish Peroxide

SummaryHistochemical methods were adapted to study the passage of different sized molecules through the aortic wall of rats and rabbits. Horseradish peroxidase, myoglobin and cytochrome C served as tracers of different molecular weights easily identifiable by light microscopy after peroxidase staining. All three proteins were transported from the lumen through the aortic wall to the adventitia; no penetration from the adventitia was seen. When compared for rate of transportation, horseradish peroxide with the highest molecular weight was slower than myoglobin and cytochrome C. In the vena cava the transport of these proteins occurred from both directions but the media was saturated from the lumen only. No difference between velocity of transport of horseradish peroxidase, myoglobin and cytochrome C was found in the vena cava. In vitro incubation in various solutions containing dissolved horseradish peroxidase showed penetration of peroxidase to be minimal. The molecular weight of material transported and pressure in the lumen are considered as the most important factors influencing the transport of nutrients through the wall of an avascular large vessel. When abdominal aorta was isolated from surrounding tissue, clearance of peroxidase from the vessel wall was greatly retarded.

scholarly journals Optimization of Laccase from Ganoderma lucidum Decolorizing Remazol Brilliant Blue R and Glac1 as Main Laccase-Contributing Gene

Molecules ◽  
2019 ◽  
Vol 24 (21) ◽  
pp. 3914 ◽  
Author(s):  
Peng Qin ◽  
Yuetong Wu ◽  
Bilal Adil ◽  
Jie Wang ◽  
Yunfu Gu ◽  
...  
Keyword(s):  
Ganoderma Lucidum ◽  
Reactive Dye ◽  
White Rot ◽  
Brilliant Blue ◽  
White Rot Fungus ◽  
Protein Mass ◽  
Remazol Brilliant Blue R ◽  
Protein Mass Spectrometry ◽  
Laccase Enzyme

Many dyes and pigments are used in textile and printing industries, and their wastewater has been classed as a top source of pollution. Biodegradation of dyes by fungal laccase has great potential. In this work, the influence of reaction time, pH, temperature, dye concentration, metal ions, and mediators on laccase-catalyzed Remazol Brilliant Blue R dye (RBBR) decolorization were investigated in vitro using crude laccase from the white-rot fungus Ganoderma lucidum. The optimal decolorization percentage (50.3%) was achieved at 35 °C, pH 4.0, and 200 ppm RBBR in 30 min. The mediator effects from syringaldehyde, 1-hydroxybenzotriazole, and vanillin were compared, and 0.1 mM vanillin was found to obviously increase the decolorization percentage of RBBR to 98.7%. Laccase-mediated decolorization percentages significantly increased in the presence of 5 mM Na+ and Cu2+, and decolorization percentages reached 62.4% and 62.2%, respectively. Real-time fluorescence-quantitative PCR (RT-PCR) and protein mass spectrometry results showed that among the 15 laccase isoenzyme genes, Glac1 was the main laccase-contributing gene, contributing the most to the laccase enzyme activity and decolorization process. These results also indicate that under optimal conditions, G. lucidum laccases, especially Glac1, have a strong potential to remove RBBR from reactive dye effluent.

scholarly journals Adsorption of Remazol Brilliant Blue R Using Amino-Functionalized Organosilane in Aqueous Solution

2017 ◽  
Vol 17 (3) ◽  
pp. 343 ◽  
Author(s):  
Ozi Adi Saputra ◽  
Alifia Harista Rachma ◽  
Desi Suci Handayani
Keyword(s):  
Adsorption Process ◽  
Morphological Structure ◽  
Brilliant Blue ◽  
Kinetic Rate ◽  
Isotherm Study ◽  
Remazol Brilliant Blue R ◽  
Infra Red ◽  
Optimum Ph ◽  
Isotherm Adsorption ◽  
Pseudo Second Order

Synthesis of amino functionalized organosilane (AFOS) via UV-irradiation method has been conducted. In this research, the precursors for synthesizing AFOS are 3-glycydiloxypropyl trimethoxysilane (GPTMS) and ethylene diamine. Synthesis of amino functionalized organosilane was performed for 16 h monitored by Infra-red spectroscopy technique. The morphological structure of AFOS was observed by scanning electron microscopy. The adsorption process was conducted in the batch method using Remazol Brilliant Blue R (RBBR) as anionic dye model. In this study, the pH, contact time and dyes concentration were varied to determine the optimum pH, kinetic and isotherm adsorption. Based on the calculation, the kinetic rate of the RBBR adsorbed onto AFOS was determined by k2 (pseudo second-order). Moreover, the isotherm study showed that the Langmuir model fitted for the adsorption of RBBR onto AFOS with Qm by 21.3 mg g-1.

To the Mechanism of 2-Nitroanisole Carcinogenicity: in vitro Metabolism of 2-Nitroanisole Mediated by Peroxidasesand Xanthine Oxidase

1998 ◽  
Vol 63 (6) ◽  
pp. 857-869 ◽  
Author(s):  
Marie Stiborová ◽  
Heinz H. Schmeiser ◽  
Eva Frei
Keyword(s):  
Horseradish Peroxidase ◽  
Xanthine Oxidase ◽  
Dna Adducts ◽  
Active Oxygen Species ◽  
Radical Mechanism ◽  
Maximal Velocity ◽  
Prostaglandin H Synthase ◽  
Nuclease P1 ◽  
Prostaglandin H

The in vitro enzymatic metabolism of carcinogenic 2-nitroanisole was investigated using peroxidases (horseradish peroxidase and prostaglandin H synthase) and xanthine oxidase catalyzing oxidative and reductive reactions, respectively. The oxidation of 2-nitroanisole catalyzed by horseradish peroxidase exhibits the Michaelis-Menten kinetics. The Michaelis constant (Km) and the maximal velocity (Vmax) values for this substrate were determined at pH 5.0, 7.0, 7.6 and 8.0. At optimal pH (7.6), the Km and Vmax values are 0.219 μmol/l and 34.45 pmol/min per nmol peroxidase, respectively. The oxidation of 2-nitroanisole is inhibited by radical trapping agents (NADH, ascorbate, glutathione and nitrosobenzene). This indicates that the peroxidase-mediated oxidation of 2-nitroanisole proceeds via a radical mechanism. Active oxygen species are formed during the horseradish peroxidase-catalyzed reactions in the presence of NADH, hydrogen peroxide and 2-nitroanisole. 2-Nitroanisole is also oxidized by mammalian prostaglandin H synthase. Using the nuclease P1-enhanced variation of the 32P-postlabelling assay, the formation of DNA adducts was detected in DNA treated with 2-nitroanisole and xanthine oxidase. No DNA binding was detected after oxidation of 2-nitroanisole with horseradish peroxidase and prostaglandin H synthase. The results presented (the formation of DNA adducts after 2-nitroanisole activation by xanthine oxidase and that of radicals and/or superoxide radicals during the reactions with peroxidases) strongly suggest the participation of 2-nitroanisole both in the initiation and in the promotion phases of carcinogenesis.

scholarly journals Influence of pH on the growth, laccase activity and RBBR decolorization by tropical basidiomycetes

2009 ◽  
Vol 52 (5) ◽  
pp. 1075-1082 ◽  
Author(s):  
Sérgio Luiz Moreira Neto ◽  
Dácio Roberto Matheus ◽  
Kátia Maria Gomes Machado
Keyword(s):  
Culture Medium ◽  
Laccase Activity ◽  
Fungal Growth ◽  
Brilliant Blue ◽  
Enzymatic System ◽  
Industrial Residues ◽  
Optimum Ph ◽  
Influence Of Ph ◽  
Basidiomycete Fungi

The basidiomycete fungi Lentinus crinitus and Psilocybe castanella are being evaluated in a bioremediation process of soils contaminated with organochlorine industrial residues in the Baixada Santista, São Paulo. The aim of the present study was to determine the influence of pH on the fungal growth, in vitro decolorization of anthraquinonic dye Remazol Brilliant Blue R (RBBR) and laccase activity. The pH of the culture medium influenced the growth of L. crinitus and P. castanella, which presented less growth at pH 5.9 and pH 2.7, respectively. The fungi were able to modify the pH of the culture medium, adjusting it to the optimum pH for growth which was close to 4.5. Decolorization of the RBBR was maximal at a pH of 2.5 to 3.5. Higher laccase activity was observed at pH 3.5 and pH 4.5 for L. crinitus and P. castanella, respectively. pH was found to be an important parameter for both the growth of these fungi and the enzymatic system involved in RBBR decolorization.

Biosynthetic Enzymes of Tetrahydrolimipterin from Green Sulfur Bacterium Chlorobium Limicola

Pteridines ◽  
1998 ◽  
Vol 9 (2) ◽  
pp. 69-84 ◽  
Author(s):  
Dongmin Kang ◽  
Sangjoon Kim ◽  
Jeongbin Yim
Keyword(s):  
Molecular Weight ◽  
Gtp Cyclohydrolase I ◽  
Chlorobium Limicola ◽  
Gtp Cyclohydrolase ◽  
Subunit Molecular Weight ◽  
Substrate Analogues ◽  
Sds Page ◽  
Optimum Ph ◽  
Green Sulfur

Summay Based on the structure of limipterin (Cha, Pfleiderer, and Yim, Helv, Chim. Acta, 78: 600-614. 1995 the biosynthetic pathway for the newly identified pterin glycoside was investigated. It was demonstrated that tetrahydrolimipterin (H4 -limipterin) can be synthesized from GTP by the enzymes GTP cyclohydrolase I, 6-pyruvoyltetrahydropterin (PTP) synthase, sepiaptcrin reductase and limipterin synthase, present in the extract of Chlorobium limicola. Limipterin synthase (UDP-N -acetylglucosamine:5 ,6,7 ,8-tetrahydro-L-biopterin 2'-O-β-N-acetylglucosaminyl transferase) catalyzed the condensation of tetrahydrobiopterin (H4-biopterin) with UDP-N-acetylglucosamine in the presence of dithiothreitol and MnCl2. It could also produce D-tepidopterin, [1-O-(D-threo- biopterin-2'-yl)- β-N -acetylglucosamine] when 5,6,7 ,8-tetrahydro-D-threobiopterin and UDP-N-acetylglucosamine were used as substrates . Substrate analogues such as UTP, UDP and UDP-Nacetylgalactosamine inhibit the enzyme activity. The Km values for tetrahydrobiopterin and UDP-N-acetylglucosamine were 42.2 μM and 124.3 μM, respectively. Optimum pH and temperature were pH 8.0 and 37°C. The molecular weight of the enzyme was calculated to be 46,300 dalton from a calibrated Superdex 75 and the subunit molecular weight was estimated at 46,000 dalton by SDS-PAGE. These results suggest that limipterin synthase exists as a monomer. Biosynthetic intermediates of H4-limipterin such as N2NTP , 6-PTP, and H4-biopterin were identified in vitro using purified GTP cyclohydrolase I, PTP synthase, sepiapterin reductase, and limipterin synthase. From the HPLC and TLC analyses of the enzymic intermediater, it could be concluded that H4-limipterin comes from GTP by way of H4-biopterin in Chlorobium limicola.

Dermatan Sulfate and LMW Heparin Enhance the Anticoagulant Action of Activated Protein C

1999 ◽  
Vol 82 (11) ◽  
pp. 1462-1468 ◽  
Author(s):  
José Fernández ◽  
Jari Petäjä ◽  
John Griffin
Keyword(s):  
Molecular Weight ◽  
Unfractionated Heparin ◽  
Protein C ◽  
Dermatan Sulfate ◽  
Anticoagulant Activity ◽  
Activated Protein C ◽  
Factor V ◽  
Factor Va ◽  
Anticoagulant Action

SummaryUnfractionated heparin potentiates the anticoagulant action of activated protein C (APC) through several mechanisms, including the recently described enhancement of proteolytic inactivation of factor V. Possible anticoagulant synergism between APC and physiologic glycosaminoglycans, pharmacologic low molecular weight heparins (LMWHs), and other heparin derivatives was studied. Dermatan sulfate showed potent APC-enhancing effect. Commercial LMWHs showed differing abilities to promote APC activity, and the molecular weight of LMWHs correlated with enhancement of APC activity. Degree of sulfation of the glycosaminoglycans influenced APC enhancement. However, because dextran sulfates did not potentiate APC action, the presence of sulfate groups per se on a polysaccharide is not sufficient for APC enhancement. As previously for unfractionated heparin, APC anticoagulant activity was enhanced by glycosaminoglycans when factor V but not factor Va was the substrate. Thus, dermatan sulfate and LMWHs exhibit APC enhancing activity in vitro that could be of physiologic and pharmacologic significance.

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