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(12) United States Patent (10) Patent No.: US 8,778,643 B2 Franz Et Al

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(12) United States Patent (10) Patent No.: US 8,778,643 B2 Franz Et Al US008778643B2 (12) United States Patent (10) Patent No.: US 8,778,643 B2 Franz et al. (45) Date of Patent: Jul. 15, 2014 (54) METHODS FOR INCREASING LIPID LEVELS Baguz et al., “Synergistic Effect of Auxins and Brassinosteroids on AND PRODUCING TRACYLGLYCEROLS IN the Growth and Regulation of Metabolite Content in the Green Alga ALGAE Chlorella vulgaris (Trebouxiophyceae). Plant Physiology and Bio chemistry, vol. 71, 2013, pp. 290-297. (71) Applicant: The Regents of the University of Bishop et al., “The Effect of Chloramphenicol and Cycloheximide on California, Oakland, CA (US) Lipid Synthesis during Chloroplast Development in Euglena gracilis”, Archives of Biochemistry and Biophysics, vol. 137, 1970, (72) Inventors: Annaliese Kirsten Franz, Davis, CA pp. 179-189. (US); Megan Danielewicz, Davis, CA Burns et al., “High-Throughput Screening of Small Molecules for (US); Diana Meiying Wong, Davis, CA Bioactivity and Target Identification in Caenorhabditis elegans', (US); Lisa Adele Anderson, Davis, CA Nature Protocols, vol. 1, No. 4, 2006, pp. 1906-1914. (US) Chen et al., “A High Throughput Nile Red Method for Quantitative Measurement of Neutral Lipids in Microalgae'. Journal of Micro (73) Assignee: The Regents of the University of biological Methods, vol. 77, 2009, pp. 41-47. California, Oakland, CA (US) Dehesh et al., “Overexpression of 3-Ketoacyl-Acyl-Carrier Protein Synthase Ills in Plants Reduces the Rate of Lipid Synthesis'. 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Oil-Producing Alga Nannochloropsis Sp., PNAS, vol. 108, No. 52, Dec. 27, 2011, pp. 21265-21269. (56) References Cited Kim et al., “Combinatorial Solid-Phase Synthesis of 4,6-Diaryland U.S. PATENT DOCUMENTS 4-Aryl, 6-Alkyl-1,3,5-triazines and their Application to Efficient Biofuel Production'. ACS Combinatorial Science, vol. 14, 2012, pp. 7,901928 B2 3/2011 Yadav et al. 395-398. 2011, 0078946 A1 4/2011 Newell et al. (Continued) FOREIGN PATENT DOCUMENTS WO 2010, 132413 A1 11, 2010 Primary Examiner — Ralph Gitomer (74) Attorney, Agent, or Firm — Morrison & Foerster LLP OTHER PUBLICATIONS El-Baky H. et al. Evaluation of Marine Alga Ulva lactuca L. as a (57) ABSTRACT Source of Natural Preservative Ingredient. American Eurasian JAgri culture & Environmental Science 3(3)434-444, 2008.* The present disclosure relates to methods of increasing lipid Mostafa S. Microalgal Biotechnology: Prospects and Applications levels in an algal or yeast cell population, methods of produc Chapter 12, published by Intech, 2012.* ing Saturated or monounsaturated triacylglycerols in an algal Herrero M. etal. Sub and Supercritical Fluid Extraction of Functional Ingredients from Different Natural Sources . Instituto de or yeast cell population, and methods of decreasing polyun Fermentaciones Industriales Madrid Spain.* saturated triacylglycerol production in an algal or yeast cell International Search Report and Written Opinion received for PCT population by contacting the cell population with a chemical Patent Application No. PCT/US2013/031059, mailed on Jun. 28, compound that is capable of increasing lipid levels or altering 2013, 10 pages. the lipid composition in the cell population. Avasthi et al., “A Chemical Screen Identifies Class A G-Protein Coupled Receptors AS Regulators of Cilia'. ACS Chemical Biology, vol. 7, 2012, pp. 911-919. 17 Claims, 24 Drawing Sheets US 8,778,643 B2 Page 2 (56) References Cited Ryckebosch et al., "Optimization of an Analytical Procedure for Extraction of Lipids from Microalgae'. Journal of the American Oil OTHER PUBLICATIONS Chemists Society, Jul. 24, 2011, 10 pages. Saffari et al., “Green Tea Metabolite EGCG Protects Membranes Against Oxidative Damage invitro'. Life Sciences, vol. 74,2004, pp. Lambert et al., “Mechanisms of Cancer Prevention by Tea Constitu 1513-1518. ents'. Proceedings of the Third International Scientific Symposium Schimmel, Richard J., “Stimulation of cAMP Accumulation and on Tea and Human Health: Role of Flavonoids in the Diet, The Lipolysis in Hamster Adipocytes with Forskolin'. The American Journal of Nutrition, vol. 133, 2003, pp. 3262S-3267S. Physiological Society, 1984, pp. C63-C68. 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