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.
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