Development of designer transgenic bacteria and/or cyanobacteria for production of advanced biofuels such as butanol and other higher alcohols is a significant area of renewable energy research. A rapid and accurate analytical method to measure biofuel butanol and other higher alcohols in liquid media of microbial culture is highly desirable in this arena. This experimental study innovatively applied a well-established alcohol colorimetric assay system with a somewhat promiscuous alcohol dehydrogenase (ADH) and a coloring reagent to detect and quantify butanol from the liquid culture of genetically transformed E. coli strain that contains a set of designer butanol-production-pathway genes. The colorimetric butanol quantification was made possible by ADH's broad substrate specificity, which allows for the oxidation of butanol to convert NAD+ to NADH.  A standard colorimetric curve that was generated with a series of butanol solutions with known concentrations can be used in determining the butanol concentration in the genetically transformed E. coli culture.  The liquid E. coli culture can then be subjected to a broad qualitative analysis via a 200 amu Universal Gas Analyzer to confirm the identity of the analyte alcohol species. The colorimetric assay in combination with the techniques of multiwells microplate reader may be used to explore the applicability of quantifying other higher alcohols such as pentanol or hexanol as well.

Colorimetric assay for quantitative analysis of butanol and other higher alcohols

246th National Meeting (2013)

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