Team:Washington/Alkanes/Results

From 2011.igem.org

GC-MS analysis shows that our alkane production system is working

We proformed GC-MS analysis on cell cultures expressing both Acyl-ACP reductase(AAR) and Aldehyde Decarbonylase(ADC). to act as controls and in order to show that the alkane production system is working as expected.

GC plot showing that alkanes are only produced when both ADC and AAR are expressed.

No significant hydrocarbons peaks in samples extracted from cells expressing only ADC

No significant peaks are found on the ADC-Only control GC plot within the time range that we expect alkane signal based upon GC-MS runs on chemical standards(8-10.5 minutes). This is expected, as E. coli does not normally produce any long chain length aldehyde substrates.

C16-Alcohol production by cells expressing AAR

The GC plot from the extraction from a cell culture expressing only AAR shows a significant peak at 10.2 minutes corresponding to the C16 alcohol( as confirmed by comparison of the peak's MS spectra to a reference library). E. coli produces multiple aldehyde dehydrogenases that convert aldehydes to alcohols. The production of C16 alcohols in cells expressing AAR( but not in cells expressing only ADC) is consistent with AAR reducing even chain length Acyl-ACPs into even chain length fatty aldehydes, which are further reduced by aldehyde dehydrogenases to the alcohol.