Microplate assay for endo-polygalacturonase activity determination based on ruthenium red method
2014
Ortiz, Gastón E. | Guitart, María E. | Alberto, Edgardo | Fernández Lahore, Héctor M. | Blasco, Martin
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, 40μg of polygalacturonic acid, and 45μg of RR for each sample determination. Furthermore, we obtained an interconversion factor for RR and viscosity activities.
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