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Colorimetric Endpoint Assay for Enzyme-catalyzed Iodide Ion Release for High-throughput Screening in Microtiter Plates by Sanela Kurtovic; Ronnie Jansson; Bengt Mannervik is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Colorimetric Endpoint Assay for Enzyme-catalyzed Iodide Ion Release for High-throughput Screening in Microtiter Plates about?

Efforts are being made to engineer enzymes with enhanced activities against haloalkanes, a toxicologically important class of compounds widely used and frequently occurring in the environment. Here we describe a facile, inexpensive, and robust method for the screening of libraries of mutated enzymes with iodoalkane substrates. Iodide formed in the enzymatic reaction is oxidized to iodine, which in the presence of starch gives blue color that can be measured at 610 nm or scored with the human eye. The assay can be performed with enzymes in crude cell lysates in 96-wells microtiter plates. Expression clones of several glutathione transferases showed diverse activities with different iodoalkanes, and a mutant library of human glutathione transferase A1-1 expressed variants with enhanced substrate selectivities.

Who reads Colorimetric Endpoint Assay for Enzyme-catalyzed Iodide Ion Release for High-throughput Screening in Microtiter Plates?

Author

Sanela Kurtovic; Ronnie Jansson; Bengt Mannervik

Publisher

Elsevier BV

Published

2007

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)