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Team:Nevada/Notebook/June/Week2/2
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E. Coli Cyano Enzymology Media
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Assay for ethanol detection. Simple one-step assay takes ethanol and NAD+ into
acetylaldahyde and NADH with use of alcohol Deh (ADH) enzyme. Final NADH concentration
can be determined using its molar extinction coefficient and absorbance at 340.0 nm, and
is proportionate to initial ethanol concetration.
Ethanol concentrations ranging from 0.05mM- 0.25mM were assayed with 173U/mL ADH.
and absorbances were measured at 340.0 nm.
Results: Final NADH concentrations were calculated to be off by a factor of ten due
to unexpectedly low absorbancies. Assay was repeated with increased ethanol concentrations
(0.5-10mM), but absorbancies remained lower
than expected.
Disscussion: It was determined that this assay was now sensitive enough for the range of
ethanol concentrations that will need to be detected in our project, therefore we will use
other means of ethanol detection using simple primary alcohol detection methods using
oxidizing reagents.
Media
