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Detection of alkanes, alcohols, and aldehydes using bioluminescence
Author(s) -
MinakBernero Vera,
Bare Richard E.,
Haith Copper E.,
Grossman Matthew J.
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20089
Subject(s) - alcohol dehydrogenase , aldehyde , alkane , luciferase , chemistry , bioluminescence , alcohol oxidation , alcohol , aldehyde dehydrogenase , chemiluminescence , primary alcohol , catalysis , biochemistry , enzyme , organic chemistry , gene , transfection
We report a novel method for the rapid, sensitive, and quantitative detection of alkanes, alcohols, and aldehydes that relies on the reaction of bacterial luciferase with an aldehyde, resulting in the emission of light. Primary alcohols with corresponding aldehydes that are within the substrate range of the particular luciferase are detected after conversion to the aldehyde by an alcohol dehydrogenase. In addition, alkanes themselves may be detected by conversion to primary alcohols by an alkane hydroxylase, followed by conversion to the aldehyde by alcohol dehydrogenase. We developed a rapid bioluminescent method by genetically engineering the genes encoding bacterial luciferase, alcohol dehydrogenase, and alkane hydroxylase into a plasmid for simultaneous expression in an E. coli host cell line. Alkanes, alcohols, or aldehydes were detected within seconds, with sensitivity in the micromolar range, by measuring the resulting light emission with a microplate reader. We demonstrate the application of this method for the detection of alkanes, alcohols, and aldehydes and for the detection of alkane hydroxylase and alcohol dehydrogenase activity in vivo. This method is amenable to the high‐throughput screening needs required for the identification of novel catalysts. © 2004 Wiley Periodicals, Inc.