Bioengineered protein phosphatase 2A

Instrumental Methods for Detection of Lipophilic Marine Toxins in Endemic Species from Pacific Austral Fjords

“…The PP2a inhibition assay (PP2aIA) is a rapid method for the detection of toxins associated with the OA-group, which is based on its functional property of inhibiting type PP2a (Figure 7) [112][113][114]. In general, this method tends to be precise, sensitive, reproducible, simple, and fast [47,115,116].…”

“…Several methods of purification of PP2a have been proposed, including: purification of recombinant PP2a overexpressed in insect cells [114,117,118], PP2ac expression in mammalian cells [119], and overexpression of PP2ac in yeast [120]. However, none of these systems is capable of producing a high yield of recombinant proteins [110,121].…”

New biorecognition molecules in biosensors for the detection of toxins

“…Various constructs of the PP2A inhibition-based biosensor have been reported for electrochemical monitoring of MC-LR in cyanobacterial cell samples (Campàs et al, 2005(Campàs et al, , 2007b or for okadaic acid in marine samples (Volpe et al, 2009;Campàs and Marty, 2007c). In the same context, Rubiolo et al (2013), genetically modified the PP2A catalytic site to enhance the enzyme stability and activity.…”

“…On the other side, the enzyme in its active form was successfully expressed in yeast, mammalian cells (Swiatek et al, 2000), insect cells (Myles et al, 2001) and insect larvae (Rubiolo et al, 2012). Very low quantities of active enzyme were produced in yeast and mammalian cells, while a high expression level was achieved in insect cells and insect larvae (Rubiolo et al, 2013). Both systems, insect cell driven expression and insect larvae expression were characterized with high quantities of the catalytic subunit of PP2A with enhance activity and inhibition by okadaic acid (OA), similar to that observed for the enzyme purified from animal tissues (Rubiolo et al, 2012).…”