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Heterologous expression of maize Fatty Acid Elongase (FAE) components in yeast - growth analysis

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posted on 26.03.2019 by Alexis Campbell, Kenna Stenback, Basil J. Nikolau, M. Ann Perera, Trang Hoang
Fatty acids of more than 18-carbons, generally known as very long chain fatty acids (VLCFAs) are essential for eukaryotic cell viability, and uniquely in terrestrial plants they are the precursors of the cuticular lipids that form the organism’s outer barrier to the environment. VLCFAs are synthesized by fatty acid elongase (FAE), which is an integral membrane enzyme system with multiple components. The genetic complexity of the FAE system, and its membrane association has hampered the biochemical characterization of FAE. In this study we computationally identified Zea mays genetic sequences that encode the enzymatic components of FAE and developed a heterologous expression system to evaluate their functionality. The ability of the maize components to genetically complement Saccharomyces cerevisiae lethal mutants confirmed the functionality of ZmKCS4, ZmELO1, ZmKCR1, ZmKCR2, ZmHCD and ZmECR, and the VLCFA profiles of the resulting strains were used to infer the ability of each enzyme component to determine the product profile of FAE.

In addition to very long chain fatty acid data, strain growth measurements were taken by measuring cell density at Optical Density (OD) 600 nm. Measurements were taken on a time course for each of the strains analyzed. For additional information please see code book.

This data supports the article:
Campbell AA, Stenback KE, Flyckt K, Hoang T, Perera MAD, et al. (2019) A single-cell platform for reconstituting and characterizing fatty acid elongase component enzymes. PLOS ONE 14(3): e0213620.

Funding

Center for Metabolic Biology (www.metabolicbiology.iastate.edu)

Metabolomics: Advancing the Scientific Promise to Better Understand Plant Specialized Metabolism for a Low-Carbon Society

Directorate for Biological Sciences

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NSF Engineering Reasearch Center for Biorenewable Chemicals (CBiRC)

Directorate for Engineering

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NSF Supplemental awards 1456245 and 1555457

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