Functional Characterization of Fatty Acyl-Reductase CG17560 in Drosophila Melanogaster

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Authors

Yi, Bo R.

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2014

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Thesis

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en_US

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Over the past few decades, there has been growing interest in biofuel development to supplement current energy sources, with different approaches being explored. Use of enzymes from insects, such as fatty-acyl reductases (FARs), has been one such approach. FARs convert fatty acids to equivalent aldehydes and/or alcohols. Aldehydes may serve as precursors for decarbonylation, resulting in hydrocarbon production which could then be purified as biofuel. The FAR 17560 protein from Drosophila melanogaster was analyzed for its substrate specificity and resulting product. The experiment involved baculoviral expression of the FAR 17560 gene in SF9 cells and functional assays with various free fatty acid substrates, using whole cell incubations. Initial reactions with infected cells producing recombinant FAR 17560 showed uptake of substrate when given a free fatty acid cocktail (24:0, 26:0, 28:0) with only 24:0 being taken up in preferentially, however aldehyde or alcohol products from these substrates were not detected . The free fatty acid 18:0 was also tested for substrate specificity. These data suggest that FAR 17560 may have specificity for 24:0 FFA and smaller chain length FFAs. This suggests that FARs work together with other enzymes not present in the SF9 cells, which would explain why only FFA uptake without substrate modification was observed. As such, conducting additional research can help to further identify the specificity of FAR 17560 protein.

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