Microplate assay for endo-polygalacturonase activity determination based on ruthenium red method

Endo-polygalacturonase (endo-PGase) activity determinations generally rely on viscosity changes or reducing sugar ends produced by this activity over polygalacturonic acid. Torres and coworkers [Enzyme Microb. Technol. 48 (2011) 123–128] showed that ruthenium red (RR) is useful for endo-PGase determ...

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Detalhes bibliográficos
Autores: Ortiz, Gastón Ezequiel, Guitart, María Eugenia, Alberto, Edgardo Omar, Fernandez Lahore, Hector, Blasco, Martín
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2014
País:Argentina
Recursos:Consejo Nacional de Investigaciones Científicas y Técnicas
Repositorio:CONICET Digital (CONICET)
Idioma:inglés
OAI Identifier:oai:ri.conicet.gov.ar:11336/32548
Acesso em linha:http://hdl.handle.net/11336/32548
Access Level:acceso abierto
Palavra-chave:Microplate
Pectinases
Endo-Polygalacturonases
Assay
Ruthenium Red
High Throughput
https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
Descrição
Resumo:Endo-polygalacturonase (endo-PGase) activity determinations generally rely on viscosity changes or reducing sugar ends produced by this activity over polygalacturonic acid. Torres and coworkers [Enzyme Microb. Technol. 48 (2011) 123–128] showed that ruthenium red (RR) is useful for endo-PGase determination. In this article, we present a high-throughput liquid-based endo-PGase assay based on the RR method and compare it with the viscosity determination method. The reduced assay uses a small volume of enzyme solution, 40 μg of polygalacturonic acid, and 45 μg of RR for each sample determination. Furthermore, we obtained an interconversion factor for RR and viscosity activities.