Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 110, Issue 49, Pages 19748-19753Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1309299110
Keywords
metabolism; RNAi; algal biofuel; targeted manipulation; triacylglycerol
Categories
Funding
- National Institutes of Health Marine Biotechnology Training Grant Fellowship [5T32GM067550]
- California Energy Commission's California Initiative for Large Molecule Sustainable Fuels [500-10-039]
- Air Force Office of Scientific Research [FA9550-08-1-0178]
- Department of Energy [DE-EE0001222, DE-EE0003373]
- National Science Foundation [CBET-0903712]
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [0903712] Funding Source: National Science Foundation
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Biologically derived fuels are viable alternatives to traditional fossil fuels, and microalgae are a particularly promising source, but improvements are required throughout the production process to increase productivity and reduce cost. Metabolic engineering to increase yields of biofuel-relevant lipids in these organisms without compromising growth is an important aspect of advancing economic feasibility. We report that the targeted knockdown of a multifunctional lipase/phospholipase/acyltransferase increased lipid yields without affecting growth in the diatom Thalassiosira pseudonana. Antisense-expressing knockdown strains 1A6 and 1B1 exhibited wild-type-like growth and increased lipid content under both continuous light and alternating light/dark conditions. Strains 1A6 and 1B1, respectively, contained 2.4- and 3.3-fold higher lipid content than wild-type during exponential growth, and 4.1- and 3.2-fold higher lipid content than wild-type after 40 h of silicon starvation. Analyses of fatty acids, lipid classes, and membrane stability in the transgenic strains suggest a role for this enzyme in membrane lipid turnover and lipid homeostasis. These results demonstrate that targeted metabolic manipulations can be used to increase lipid accumulation in eukaryotic microalgae without compromising growth.
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