Journal
JOURNAL OF PHYCOLOGY
Volume 48, Issue 3, Pages 580-584Publisher
WILEY-BLACKWELL
DOI: 10.1111/j.1529-8817.2012.01137.x
Keywords
biofuel; DGAT; hyperlipidemia; lipids; oil; triacylglycerol; triglyceride
Categories
Funding
- Appalachian State University-University Research Council
- Office of Student Research
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Common methods for assaying acyl-CoA:diacylglycerol acyltransferase (DGAT) enzymatic activity rely upon radiolabeled substrates or product assay. We developed a novel assay that directly quantifies endogenous DGAT activity through the use of a fluorescently labeled substrate. We performed this assay with microsomal protein, 2-(6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)hexanoyl-1-hexadecanoyl-sn-glycero-3-diacylglycerol (NBD-DAG), and oleoyl-CoA substrates. DGAT activity was analyzed in three species of algae as well as rat liver. The protocol proved to be sensitive and reliable. This assay may be used to facilitate research in the areas of biodiesel, oilseed crops, and triacylglycerol-related human pathologies.
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