2-D solid-state assay platform: a tool for screening aldehyde-releasing enzyme activity in colonies |
| |
| Authors: | Jochen Gerlach Christoph Reisinger Brigitte Pohn Helmut Schwab Ingo Klimant |
| |
| Institution: | (1) Institute of Analytical Chemistry and Radiochemistry, Graz Technical University, Technikerstrasse 4, A-8010 Graz, Austria;(2) Research Centre Applied Biocatalysis, Petersgasse 14, A-8010 Graz, Austria;(3) Institute of Molecular Biotechnology, Graz Technical University, Petersgasse 14, A-8010 Graz, Austria |
| |
| Abstract: | A micro colony chip-based platform for high-throughput screening of proton releasing or oxygen consuming enzyme reactions
was introduced recently. Here we present a new assay platform which allows screening for the release of aromatic and aliphatic
aldehydes by enzymatic activity. The assay platform consists of 4-hydrazino-7-nitrobenzofurazane (NBD-H) as an indicator embedded
in a soft, hydrophobic polymer matrix. NBD-H forms highly fluorescent products with aldehydes via hydrazone formation. The
assay was characterized for various aliphatic and aromatic aldehydes and tested with two enzymatic reactions to show its potential.
A transesterification reaction under water-free conditions with two esterase mutants and vinyl acetate as acyl donor leads
to acetaldehyde as detectable product and a reaction under aqueous conditions with threonine aldolases. The substrates used
are phenylserine, which forms benzaldehyde, and threonine, which forms acetaldehyde. |
| |
| Keywords: | : Enzyme activity screening 2D solid state assay platform high throughput |
| 本文献已被 SpringerLink 等数据库收录！ |
|
