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Saltwater Resources and CommercialAlgae ProductionStephen MayfieldSan Diego Center for Algae BiotechnologyUniversity of California, San Diego

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Cyanobacteria Microalgae MacroalgaeProduce biodiesel typelipids and biohydrogenProduce sugars forethanol or biogasWhat are the Types of Aquatic Biomass?Produce ethanolLipids and hydrogenLareefs.comTopnews.inAlgaeAny of a group of aquatic eukaryoticorganisms that carry on photosynthesisCyanobacteriaPhotosynthetic prokaryotes found in manyenvironments, often called blue-green algaeRed AlgaeGreen AlgaeBrown AlgaeCyanobacteria

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Commercial Production in Algae TodayMicroalgaeAnnualProductionWater SourceApplication &ProductPrice (U.S. $)Spirulina3000 tonnes dryweightSaltwaterHuman NutritionAnimal NutritionCosmeticsPhycobiliproteins$53 kg-1$16 mg-1Chlorella2000 tonnes dryweightFreshwater andSaltwater StrainsHuman NutritionCosmeticsAquaculture$53 kg-1$74 l-1Dunaliella salina1200 tonnes dryweightSaltwaterHuman NutritionCosmeticsB-carotene $318-$3,186 kg-1Aphanizomenonflosaquae500 tonnes dryweightFreshwaterHuman NutritionHaematocococcuspluvialis300 tonnes dryweightFreshwaterAquacultureAstaxanthin$74 l-1$10,596 kg-1Crypthecodiniumcohnii240 tonnes DHAoilSaltwaterDHA oil $64 g-1Schizochytrium 10 tonnes DHA oil Saltwater DHA oil $64 g-1Adapted from: Brennan, L. and Owende, P. (2010) Renewable and Sustainable Energy Reviews. 14: 557-577.

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Oil Content of Some Types of MicroalgaeMicroalgae Oil Content(dry wt%)BiomassYield (g/L)GrowthRate(g/L/D)Botryococcus braunii 25-75 2Chlorella sp. 28-32 116 .65Crypthecodinium cohnii 20 15Dunaliella primolecta 23 30 .41Isochrysis sp. 25-33 6 .19Nannochloris sp. 20-35 35 .41Nannochloropsis sp. 31-68 55 .44Neochlorisoleoabundans35-54 3.2 .40Phaeodactylumtricornutum20-30 42 .32Tetraselmis sueica 15-23 20 .24Chlorella.co.nzStarcentral.mbl.eduBing.com

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Productivity is not related to water typeMicroalgae Oil Content(dry wt%)BiomassYield (g/L)GrowthRate(g/L/D)Botryococcus braunii 25-75 2Chlorella sp. 28-32 116 .65Crypthecodinium cohnii 20 15Dunaliella primolecta 23 30 .41Isochrysis sp. 25-33 6 .19Nannochloris sp. 20-35 35 .41Nannochloropsis sp. 31-68 55 .44Neochlorisoleoabundans35-54 3.2 .40Phaeodactylumtricornutum20-30 42 .32Tetraselmis sueica 15-23 20 .24Chlorella.co.nzStarcentral.mbl.eduBing.com

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Transformations technologies for algae speciesSpecies/Variety Class Method Water TypeChlamydomonasreinhardtiiChlorophyceae Bombardment,Electroporation,FreshwaterDunaliella salina Chlorophyceae Electroporation,Bombardment, GlassBeadsSaltwaterHaematococcus pluvialis Chlorophyceae Bombardment FreshwaterVolvox carteri Chlorophyceae Bombardment FreshwaterCyclotella cryptica Diatom Bombardment SaltwaterCylindrotheca fusiformis Diatom Bombardment SaltwaterNavicula saprophila Diatom Bombardment SaltwaterPhaeodactylumtricornutumDiatom Bombardment SaltwaterThalassiosira pseudonana Diatom Bombardment SaltwaterAmphidinium spp. Dinoflagellate Si Carbide Whisker SaltwaterSymbiodiniummicroadriaticumDinoflagellate Si Carbide Whisker SaltwaterChlorella ellipsoidea Trebouxiophyceae Protoplast FreshwaterChlorella saccharophila Trebouxiophyceae Electroporation FreshwaterChlorella vulgaris Trebouxiophyceae Protoplast Freshwater andSaltwaterChlorella sorokiniana Trebouxiophyceae Bombardment FreshwaterPorphyra yezoensis Rhodophyta Bombardment SaltwaterEuglena gracilis Euglenaceae Bombardment Freshwater andSaltwaterPorphyridium spp. Porphyridiophyceae Bombardment Saltwater

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Robust molecular technologies areavailable for a limited set of algaeER-mCherry mCerulean-NLS2 colorFACSER-mCherry,mCerulean-NLS3 color FACSER-TS-mCherry,mCerulean-NLS,Mito-VenusXMito-VenusXA B C DE

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Engineering of the salt water strain D. tertiolectafor recombinant protein production in chloroplasts

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XylanaseTrichoderma reeseiMannasesAspergillus sulphureusPhytaseShigella spp or E. coliCaulobacter crescentusAlpha-galactosidaseLactobacillus acidophilusTrichoderma reeseiAnalysis of recombinant industrial enzymeproduction in D. tertiolecta

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1 - Wild type 5 - Galactosidase2 - Xylanase 6 - Phytase3 - Manannase 7 - Phytase4 - GalactosidaseComparison of protein accumulation betweenC. reinhardtii and D. tertiolecta1 2 3 4 5 6 7D. tertiolecta1 2 3 4 5 6 7C. reinhardtii

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Where do we go from here?Where do we go from here?