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

作者：Ortiz Gaston E; Guitart Maria E; Alberto Edgardo; Lahore Hector M Fernandez; Blasco Martin^*

来源：Analytical Biochemistry, 2014, 454: 33-35.

DOI：10.1016/j.ab.2014.02.028

• Microplate
• Pectinases
• Endo-polygalacturonases
• Assay
• Ruthenium red
• High throughput

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 mu g of polygalacturonic acid, and 45 mu g of RR for each sample determination. Furthermore, we obtained an interconversion factor for RR and viscosity activities.

• 出版日期2014-6-1