A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model

Autores
Miquet, Johanna Gabriela; Sotelo, Ana Isabel; Gonzalez, Lorena; Gonzalez-Lebrero, Rodolfo Martin
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Enzyme kinetics is an essential topic in undergraduate Biochemistry courses. A laboratory work that covers the principal basic concepts of enzyme kinetics in steady state is presented. The alkaline phosphatase catalyzed reaction of phenyl-phosphate hydrolysis was studied as a model. The laboratory experience was designed to reinforce the concepts of initial velocity dependence on substrate and enzyme concentration, and to highlight the importance of the accurate determination of initial reaction rate. The laboratory work consists in two parts, in which students first determine the enzyme concentration and the time to be used in the following session to obtain the kinetic parameters (KM and Vmax ) by non-lineal fitting of the Michaelis-Menten equation to the initial velocity dependence with substrate concentration results. The experimental methodology is robust, the cost per student is low and the equipment and reagents used are of easy access.
Fil: Miquet, Johanna Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Sotelo, Ana Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Gonzalez, Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Gonzalez-Lebrero, Rodolfo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Materia
ALKALINE PHOSPHATASE
ENZYME KINETICS
EXPERIMENTAL DESIGN
HANDS-ON LEARNING
LABORATORY EXPERIMENT
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:11336/121404
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/121404
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a modelMiquet, Johanna GabrielaSotelo, Ana IsabelGonzalez, LorenaGonzalez-Lebrero, Rodolfo MartinALKALINE PHOSPHATASEENZYME KINETICSEXPERIMENTAL DESIGNHANDS-ON LEARNINGLABORATORY EXPERIMENThttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1https://purl.org/becyt/ford/5.3https://purl.org/becyt/ford/5Enzyme kinetics is an essential topic in undergraduate Biochemistry courses. A laboratory work that covers the principal basic concepts of enzyme kinetics in steady state is presented. The alkaline phosphatase catalyzed reaction of phenyl-phosphate hydrolysis was studied as a model. The laboratory experience was designed to reinforce the concepts of initial velocity dependence on substrate and enzyme concentration, and to highlight the importance of the accurate determination of initial reaction rate. The laboratory work consists in two parts, in which students first determine the enzyme concentration and the time to be used in the following session to obtain the kinetic parameters (KM and Vmax ) by non-lineal fitting of the Michaelis-Menten equation to the initial velocity dependence with substrate concentration results. The experimental methodology is robust, the cost per student is low and the equipment and reagents used are of easy access.Fil: Miquet, Johanna Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Sotelo, Ana Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Gonzalez, Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaFil: Gonzalez-Lebrero, Rodolfo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; ArgentinaJohn Wiley & Sons Inc2019-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/121404Miquet, Johanna Gabriela; Sotelo, Ana Isabel; Gonzalez, Lorena; Gonzalez-Lebrero, Rodolfo Martin; A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model; John Wiley & Sons Inc; Biochemistry and Molecular Biology Education; 47; 1; 1-2019; 93-991539-3429CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://iubmb.onlinelibrary.wiley.com/doi/full/10.1002/bmb.21195info:eu-repo/semantics/altIdentifier/doi/10.1002/bmb.21195info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:40:42Zoai:ri.conicet.gov.ar:11336/121404instacron: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:34982025-09-29 09:40:42.822CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model
title A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model
spellingShingle A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model
Miquet, Johanna Gabriela
ALKALINE PHOSPHATASE
ENZYME KINETICS
EXPERIMENTAL DESIGN
HANDS-ON LEARNING
LABORATORY EXPERIMENT
title_short A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model
title_full A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model
title_fullStr A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model
title_full_unstemmed A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model
title_sort A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model
dc.creator.none.fl_str_mv Miquet, Johanna Gabriela
Sotelo, Ana Isabel
Gonzalez, Lorena
Gonzalez-Lebrero, Rodolfo Martin
author Miquet, Johanna Gabriela
author_facet Miquet, Johanna Gabriela
Sotelo, Ana Isabel
Gonzalez, Lorena
Gonzalez-Lebrero, Rodolfo Martin
author_role author
author2 Sotelo, Ana Isabel
Gonzalez, Lorena
Gonzalez-Lebrero, Rodolfo Martin
author2_role author
author
author
dc.subject.none.fl_str_mv ALKALINE PHOSPHATASE
ENZYME KINETICS
EXPERIMENTAL DESIGN
HANDS-ON LEARNING
LABORATORY EXPERIMENT
topic ALKALINE PHOSPHATASE
ENZYME KINETICS
EXPERIMENTAL DESIGN
HANDS-ON LEARNING
LABORATORY EXPERIMENT
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
https://purl.org/becyt/ford/5.3
https://purl.org/becyt/ford/5
dc.description.none.fl_txt_mv Enzyme kinetics is an essential topic in undergraduate Biochemistry courses. A laboratory work that covers the principal basic concepts of enzyme kinetics in steady state is presented. The alkaline phosphatase catalyzed reaction of phenyl-phosphate hydrolysis was studied as a model. The laboratory experience was designed to reinforce the concepts of initial velocity dependence on substrate and enzyme concentration, and to highlight the importance of the accurate determination of initial reaction rate. The laboratory work consists in two parts, in which students first determine the enzyme concentration and the time to be used in the following session to obtain the kinetic parameters (KM and Vmax ) by non-lineal fitting of the Michaelis-Menten equation to the initial velocity dependence with substrate concentration results. The experimental methodology is robust, the cost per student is low and the equipment and reagents used are of easy access.
Fil: Miquet, Johanna Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Sotelo, Ana Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Gonzalez, Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
Fil: Gonzalez-Lebrero, Rodolfo Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Química y Físico-Química Biológicas "Prof. Alejandro C. Paladini". Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de Química y Físico-Química Biológicas; Argentina
description Enzyme kinetics is an essential topic in undergraduate Biochemistry courses. A laboratory work that covers the principal basic concepts of enzyme kinetics in steady state is presented. The alkaline phosphatase catalyzed reaction of phenyl-phosphate hydrolysis was studied as a model. The laboratory experience was designed to reinforce the concepts of initial velocity dependence on substrate and enzyme concentration, and to highlight the importance of the accurate determination of initial reaction rate. The laboratory work consists in two parts, in which students first determine the enzyme concentration and the time to be used in the following session to obtain the kinetic parameters (KM and Vmax ) by non-lineal fitting of the Michaelis-Menten equation to the initial velocity dependence with substrate concentration results. The experimental methodology is robust, the cost per student is low and the equipment and reagents used are of easy access.
publishDate 2019
dc.date.none.fl_str_mv 2019-01
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/11336/121404
Miquet, Johanna Gabriela; Sotelo, Ana Isabel; Gonzalez, Lorena; Gonzalez-Lebrero, Rodolfo Martin; A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model; John Wiley & Sons Inc; Biochemistry and Molecular Biology Education; 47; 1; 1-2019; 93-99
1539-3429
CONICET Digital
CONICET
url http://hdl.handle.net/11336/121404
identifier_str_mv Miquet, Johanna Gabriela; Sotelo, Ana Isabel; Gonzalez, Lorena; Gonzalez-Lebrero, Rodolfo Martin; A laboratory work to introduce Biochemistry undergraduate students to basic enzyme kinetics -alkaline phosphatase as a model; John Wiley & Sons Inc; Biochemistry and Molecular Biology Education; 47; 1; 1-2019; 93-99
1539-3429
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://iubmb.onlinelibrary.wiley.com/doi/full/10.1002/bmb.21195
info:eu-repo/semantics/altIdentifier/doi/10.1002/bmb.21195
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
application/pdf
dc.publisher.none.fl_str_mv John Wiley & Sons Inc
publisher.none.fl_str_mv John Wiley & Sons Inc
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
_version_ 1844613287831928832
score 13.070432

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