info:eu-repo/semantics/conferencePoster
High-throughput screening method for alkaline lipases using triolein as substrate
Autor
Jorge Alberto Rodriguez Gonzalez
Institución
Resumen
Lipases (EC 3.1.1.3) are soluble enzymes
that can act on natural insoluble triacylglycerols (TAG) (1).
They are present in many bacteria, fungi, plants, and
animals, and have been used in numerous applications
such as food, cosmetics, detergents, and pharmaceutical
industries (2). Most lipases have been reported to be
optimally active at neutral pH (3), but it will be useful to
have alkaline lipases for biocatalysis and industrial
applications. The success of their screening depends on
reliable, appropriate, and sensitive assays to detect a true
lipase activity. High-throughput screening (HTS) methods
for lipases are usually performed using non appropriate
synthetic chromogenic substrates (4); new HTS methods
employing natural substrates of lipases are necessary.
In this work we propose a HTS method for alkaline lipases
using triolein (an insoluble TAG), wherein the lipolytic
activity can be measured spectrophotometrically due to
the fatty acid liberation and the color change of a pH
indicator.