A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine

Autores
Liffourrena, Andres Sebastian; Boeris, Paola Sabrina; Salvano, M. A.; Lucchesi, Gloria Ines
Año de publicación
2009
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
This article describes a simple fluorescent method for the determination of tetradecyltrimethylammonium monooxygenase (TTAB-monooxygenase) activity involving N‑dealkylation of tetradecyltrimethylammonium bromide with concomitant production of trimethylamine (TMA). Activity was determined by measuring the formation of TMA using the morin reagent and aluminium (Al). Morin reacts with Al to form a fluorescent complex, Al-morin. In the presence of TMA, Al is tightly associated with TMA, and cannot be sequestered by morin, thus providing evidence for formation of the Al-TMA complex. The concentration of TMA is estimated by calibration graphs constructed by plotting the fluorescence intensity of the Al-morin complex versus TMA concentration. The fluorescence intensities of the Al-morin complexes quenched by TMA are linearly dependent on both the time of the TTAB-monooxygenase reaction and the amount of protein used in the reaction. The kinetic behaviour is characterized by K0.5 = 4.26 x 10-4 M, and the apparent Hill coefficient (napp) = 2.24. These values are both comparable to those determined by GC-MS (K0.5 = 4.41 x 10-4 M and napp = 2.35). Advantages of this assay include rapid and efficient implementation and potential employment for routine accurate determinations of TTAB-monooxygenase activity over a wide range of substrate concentrations.
Fil: Liffourrena, Andres Sebastian. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Boeris, Paola Sabrina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Salvano, M. A.. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina
Fil: Lucchesi, Gloria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina
Materia
TETRADECYLTRIMETHYLAMONNIUM MONOOXYGENASETRIMETHYLAMINE
TRIMETHYLAMINE
MORIN
AMMONIUM QUATERNARY COMPOUNDS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/240796
Acceder
id CONICETDig_df008d8a982ad4975fd8baa95c23f592
oai_identifier_str oai:ri.conicet.gov.ar:11336/240796
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamineLiffourrena, Andres SebastianBoeris, Paola SabrinaSalvano, M. A.Lucchesi, Gloria InesTETRADECYLTRIMETHYLAMONNIUM MONOOXYGENASETRIMETHYLAMINETRIMETHYLAMINEMORINAMMONIUM QUATERNARY COMPOUNDShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1This article describes a simple fluorescent method for the determination of tetradecyltrimethylammonium monooxygenase (TTAB-monooxygenase) activity involving N‑dealkylation of tetradecyltrimethylammonium bromide with concomitant production of trimethylamine (TMA). Activity was determined by measuring the formation of TMA using the morin reagent and aluminium (Al). Morin reacts with Al to form a fluorescent complex, Al-morin. In the presence of TMA, Al is tightly associated with TMA, and cannot be sequestered by morin, thus providing evidence for formation of the Al-TMA complex. The concentration of TMA is estimated by calibration graphs constructed by plotting the fluorescence intensity of the Al-morin complex versus TMA concentration. The fluorescence intensities of the Al-morin complexes quenched by TMA are linearly dependent on both the time of the TTAB-monooxygenase reaction and the amount of protein used in the reaction. The kinetic behaviour is characterized by K0.5 = 4.26 x 10-4 M, and the apparent Hill coefficient (napp) = 2.24. These values are both comparable to those determined by GC-MS (K0.5 = 4.41 x 10-4 M and napp = 2.35). Advantages of this assay include rapid and efficient implementation and potential employment for routine accurate determinations of TTAB-monooxygenase activity over a wide range of substrate concentrations.Fil: Liffourrena, Andres Sebastian. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Boeris, Paola Sabrina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Salvano, M. A.. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Lucchesi, Gloria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaAcademic Press Inc Elsevier Science2009-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/240796Liffourrena, Andres Sebastian; Boeris, Paola Sabrina; Salvano, M. A.; Lucchesi, Gloria Ines; A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine; Academic Press Inc Elsevier Science; Analytical Biochemistry; 384; 2; 1-2009; 343-3470003-2697CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0003269708006714info:eu-repo/semantics/altIdentifier/doi/10.1016/j.ab.2008.10.001info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:11:32Zoai:ri.conicet.gov.ar:11336/240796instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-10 13:11:33.215CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine
title A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine
spellingShingle A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine
Liffourrena, Andres Sebastian
TETRADECYLTRIMETHYLAMONNIUM MONOOXYGENASETRIMETHYLAMINE
TRIMETHYLAMINE
MORIN
AMMONIUM QUATERNARY COMPOUNDS
title_short A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine
title_full A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine
title_fullStr A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine
title_full_unstemmed A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine
title_sort A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine
dc.creator.none.fl_str_mv Liffourrena, Andres Sebastian
Boeris, Paola Sabrina
Salvano, M. A.
Lucchesi, Gloria Ines
author Liffourrena, Andres Sebastian
author_facet Liffourrena, Andres Sebastian
Boeris, Paola Sabrina
Salvano, M. A.
Lucchesi, Gloria Ines
author_role author
author2 Boeris, Paola Sabrina
Salvano, M. A.
Lucchesi, Gloria Ines
author2_role author
author
author
dc.subject.none.fl_str_mv TETRADECYLTRIMETHYLAMONNIUM MONOOXYGENASETRIMETHYLAMINE
TRIMETHYLAMINE
MORIN
AMMONIUM QUATERNARY COMPOUNDS
topic TETRADECYLTRIMETHYLAMONNIUM MONOOXYGENASETRIMETHYLAMINE
TRIMETHYLAMINE
MORIN
AMMONIUM QUATERNARY COMPOUNDS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv This article describes a simple fluorescent method for the determination of tetradecyltrimethylammonium monooxygenase (TTAB-monooxygenase) activity involving N‑dealkylation of tetradecyltrimethylammonium bromide with concomitant production of trimethylamine (TMA). Activity was determined by measuring the formation of TMA using the morin reagent and aluminium (Al). Morin reacts with Al to form a fluorescent complex, Al-morin. In the presence of TMA, Al is tightly associated with TMA, and cannot be sequestered by morin, thus providing evidence for formation of the Al-TMA complex. The concentration of TMA is estimated by calibration graphs constructed by plotting the fluorescence intensity of the Al-morin complex versus TMA concentration. The fluorescence intensities of the Al-morin complexes quenched by TMA are linearly dependent on both the time of the TTAB-monooxygenase reaction and the amount of protein used in the reaction. The kinetic behaviour is characterized by K0.5 = 4.26 x 10-4 M, and the apparent Hill coefficient (napp) = 2.24. These values are both comparable to those determined by GC-MS (K0.5 = 4.41 x 10-4 M and napp = 2.35). Advantages of this assay include rapid and efficient implementation and potential employment for routine accurate determinations of TTAB-monooxygenase activity over a wide range of substrate concentrations.
Fil: Liffourrena, Andres Sebastian. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Boeris, Paola Sabrina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Salvano, M. A.. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina
Fil: Lucchesi, Gloria Ines. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; Argentina
description This article describes a simple fluorescent method for the determination of tetradecyltrimethylammonium monooxygenase (TTAB-monooxygenase) activity involving N‑dealkylation of tetradecyltrimethylammonium bromide with concomitant production of trimethylamine (TMA). Activity was determined by measuring the formation of TMA using the morin reagent and aluminium (Al). Morin reacts with Al to form a fluorescent complex, Al-morin. In the presence of TMA, Al is tightly associated with TMA, and cannot be sequestered by morin, thus providing evidence for formation of the Al-TMA complex. The concentration of TMA is estimated by calibration graphs constructed by plotting the fluorescence intensity of the Al-morin complex versus TMA concentration. The fluorescence intensities of the Al-morin complexes quenched by TMA are linearly dependent on both the time of the TTAB-monooxygenase reaction and the amount of protein used in the reaction. The kinetic behaviour is characterized by K0.5 = 4.26 x 10-4 M, and the apparent Hill coefficient (napp) = 2.24. These values are both comparable to those determined by GC-MS (K0.5 = 4.41 x 10-4 M and napp = 2.35). Advantages of this assay include rapid and efficient implementation and potential employment for routine accurate determinations of TTAB-monooxygenase activity over a wide range of substrate concentrations.
publishDate 2009
dc.date.none.fl_str_mv 2009-01
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/240796
Liffourrena, Andres Sebastian; Boeris, Paola Sabrina; Salvano, M. A.; Lucchesi, Gloria Ines; A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine; Academic Press Inc Elsevier Science; Analytical Biochemistry; 384; 2; 1-2009; 343-347
0003-2697
CONICET Digital
CONICET
url http://hdl.handle.net/11336/240796
identifier_str_mv Liffourrena, Andres Sebastian; Boeris, Paola Sabrina; Salvano, M. A.; Lucchesi, Gloria Ines; A fluorescence assay for tetradecyltrimethylammonium mono-oxygenase activity that catalyzes the cleavage of the C–N bond with the production of trimethylamine; Academic Press Inc Elsevier Science; Analytical Biochemistry; 384; 2; 1-2009; 343-347
0003-2697
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0003269708006714
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.ab.2008.10.001
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Academic Press Inc Elsevier Science
publisher.none.fl_str_mv Academic Press Inc Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.993085

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