Biofuels Production & Analysis Presenter: Elmar Schmid, Ph.D. Associate Faculty, MiraCosta College, Oceanside, CA VP & CSO, Sustainable Green Technologies Inc., Escondido, CA

Why Biofuels? Biodiesel Unsaturated C16–18 Fatty Acid Methyl Esters (FAME)

National Security & Energy Portfolio Diversification America’s dependency on increasingly costly imported petroleum oil contributes to U.S. budget and trade deficit. - U.S. domestic oil production is about 8.5 million barrels/day but the nation consumes about 20 million barrels (840 million gal) of oil per day U.S. energy bill signed in 2007 mandates 36 billion gallons of fuels to come from biofuels by 2030.

Annual (2009) Oil Production/Imports of California (Thousand barrels) State’s Petroleum Oil Production is Declining California imports more than 45% of its petroleum oil from foreign sources Annual (2009) Oil Production/Imports of California (Thousand barrels) Foreign CA Domestic National Source: CA Energy Commission Annual Total: 605,089

Political Reasons & Incentives In April 2006, former CA Governor Arnold Schwarzenegger issued an executive order (S-06-06), directing Californian agencies to take steps in increasing the state’s biofuels production and biomass power generation. - 20 % of biofuels used in the state from state sources by 2010 - 40 % by 2020 and 75 % by 2050 - more electricity generation from renewable biomass. Schwarzenegger also proposed a Hydrogen Highway plan. California has invested $6.5 million to support a network of more than 16 filling stations and a growing fleet of cars and buses that run on this clean fuel.

Ecological Reasons Global average temperatures are rising most likely due to rising anthropogenic green house gas emissions (CO2, CH4, NOx) Need for “carbon neutral” fuel and energy solutions Annual carbon emissions from fossil fuel consumption Deep water horizon oil drilling rig explosion (Gulf of Mexico) April 10, 2010

Environmental Reasons Fossil fuel extraction takes increasing costly toll on diverse environments (oil spills, oil shale fracturing, mountain top mining) Accelerated fossil fuel combustion has costly negative impact on human wellbeing and health - Particulate matter (PM)  Diesel engines - NOx and ozone  Otto engines - Benzene  Gasoline - Heavy metals  Coal - SO2  Coal

What are Biofuels? Biofuels are liquid or gaseous fuels produced from renewable biomass, such as: 1. Corn  Starch  Bioethanol 2. Sugar cane  Sucrose  Bioethanol 3. Sugar beet  Sucrose  Bioethanol 4. Wood,  Cellulose  Bioethanol 5. Manure  Protein  Biogas (CH4) 6. Rapeseed  Oil  Biodiesel 7. Palm seed  Oil  Biodiesel 8. Diverse  Sugars  Biohydrogen Many biofuels are produced with the help of microorganisms, such as yeast, bacteria, or microbial enzymes

Biofuels Biogas Bioethanol Bio- H2 Biodiesel Potatoes Sugar Beet Manure Landfills Wheat Corn Sugar Cane Biogas Bioethanol Biofuels Bio- H2 Biodiesel Sun Flower Microalgae Palm Biomass Rapeseed Soybean Figure©E.Schmid-2011

Why Biofuels from Microalgae? Chlorella minutissima Photo©E.Schmid-2010

What are Microalgae? Microalgae are microscopically small aqueous life forms which perform a biological process called photosynthesis. Sun Water (H2O) Carbon Dioxide (CO2) Oxygen (O2) Light Energy + Micro algae Biomass Sugars Starch Cellulose Oils Graphic©E.Schmid-2010

Microalgae Advantages Fast growth rate and biomass productivity. Double in less than 24 hours. Higher sunlight conversion efficiency than terrestrial green plants. Solar conversion efficiency is with 4-5% a factor of 50 higher than in plants. Large scale cultivation of microalgae removes significant amounts of the green house gas CO2 from the atmosphere. Microalgae are metabolically very versatile. Many value products can can be produced, including antioxidants, poly-unsaturated fatty acids, (PUFAs), cattle feedstock and algae oils. Algae oils have been shown to be suitable for biodiesel production.

Taken from the website of Bioprodukte-Prof. Steinberg GmbH, Germany Large scale cultivation of microalgae under controlled, contamination-free conditions can be achieved in closed loop photobioreactors. Commercial tubular closed loop algae photobioreactor Taken from the website of Bioprodukte-Prof. Steinberg GmbH, Germany

Microalgae have higher oil productivity than agricultural plants Some microalgae accumulate up to 50% of their dry weight as oil. Microalgae farming has the potential to yield more than 70,000 gallons of oil per hectare per year (70,000 gal oil/ha/y). Crop Oil Yield (kg oil / ha x year) Oil Yield (gal oil / ha x year) Corn 146 45 Soybeans 375 120 Peanuts 921 282 Rapeseed/ Canola 1,000 306 Olives 1,051 322 Avocado 2,298 705 Palm oil 5,000 1,575/1,890 Algae Farming 268,950 (Valcent) 60,000 (Shell) 21,842 (Molina et al.) 33,000 (other) 82,500 18,405 6,700 10,123

Microalgae Fuel Advantage Microalgae-derived fuels offer many advantages over currently used fossil fuels: 1. Fuels can be produced in a sustainable, renewable way. - algae are harvested and quickly regrown within days or weeks within designed photobioreactor environments 2. Fuels, e.g. biodiesel, burns carbon-neutral when combusted in internal combustion engines or other energy conversion devises. - CO2 emitted through combustion of algae-derived biodiesel does not add new carbon into the atmosphere 3. Microalgae oils and fuels are non-toxic and highly bio-degradable. 4. Algae can grow in low grade water, waste water and even marine water. - Production of 1 t of algae biomass requires about 38,500 liters of water

Biofuels Production & Analysis Why a Biofuels Production & Analysis Curriculum? Photo©E.Schmid-2010

- Work force development - Economical Reasons - Work force development - Important California Biofuels Companies with rapidly developing “green tech” workforce demand in the future. Sapphire, San Diego - “green crude” from algae Verenium (former Diversa), San Diego Synthetic Genomics, San Diego - fully synthetic microbes for biofuels production Pearson Fuels, San Diego - built first ethanol (E-85) station in CA Amyris, San Francisco - genetically modified yeast for hydrocarbon production AE Biofuels, Cupertino - cellulosics ethanol production Cobalt Biofuels, Mountain View Imperial Valley Biodiesel, El Centro “In San Diego County, the algae biofuels industry already provides 410 direct jobs and $56 million in direct economic activity.”

Biofuels Production & Analysis Curriculum Lecture a. History of fossil fuels & Introduction b. Energy c. Fuels d. Carbon chemistry & Biomass e. Enzymes f. Gases g. Photosynthesis & Algae h. Biofuels Laboratory a. Media preparation & Cell counting techniques lab b. Cellulase enzyme lab c. Biohydrogen production lab d. Algae photobioreactor lab e. Algae oil extraction & analysis lab In Progress A laboratory manual and textbook written the presenter will be available as part of NBC2’s Global Biomanufacturing Curriculum at www.biomanufacturing.org/gbc

Educational biohydrogen reactor – fuel cell work station Photo©E.Schmid-2010

Educational bubble column photobioreactor work station Photo©E.Schmid-2010

$ $ Algae oil extraction & analysis curriculum ELSD- HPLC Analysis Curricular Development in Progress $ Purchase of Evaporative Light Scattering Detector ELSD- HPLC Analysis Accelerated Solvent Extraction (ASE) Transesterification Purchase of ASE Unit $ FAME (“Biodiesel”) Algae Harvest Dry Algae Biomass Algae Oil Extraction Algae Oil/FAME Analysis Figure©E.Schmid-2011

Acknowledgements Special thanks to following individuals, organizations & corporations: Mike Fino MEng – Bioprocess Technology Program, MiraCosta College, Oceanside, CA Kathleen Alfano, PhD - Director/PI, NSF ATE CREATE Renewable Energy Center, College of the Canyons, Santa Clarita, CA Sonja Wallman, PhD - Executive Director, Northeast Biomanufacturing Center and Collaborative (NBC2), Portsmouth, NH National Science Foundation (NSF) Dionex Corporation