2-D solid-state assay platform: a tool for screening aldehyde-releasing enzyme activity in colonies
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...
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| Published in | Mikrochimica acta (1966) Vol. 156; no. 3-4; pp. 209 - 218 |
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| Main Authors | , , , , |
| Format | Journal Article |
| Language | English |
| Published |
01.12.2006
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| Online Access | Get full text |
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| Summary: | 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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| Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
| ISSN: | 0026-3672 1436-5073 |
| DOI: | 10.1007/s00604-006-0640-8 |