The Bioalcohol Paradigm Fuels of the Future The Bioalcohol Paradigm yeast ethanol liquid fuel advanced biofuels phytomass fermentation lignocellulosic feedstocks synthetic biology main topics branched-chain alcohols bridge fuels energy CDC PHIL /James Gathany original slides by: Drew Sowersby (May 2011) _technical contributor for Advanced Biofuels USA www.AdvancedBiofuelsUSA.org

Message to the reader The following slide document has been created to inform a broad audience about the importance and likely dominance of bioalcohols in the transportation industry as the global transition from non-renewable fossil fuels to renewable advanced biofuels gains momentum. The information contained in these slides stands in support of the Advanced Biofuels USA mission. “The Mission of Advanced Biofuels USA is to promote public understanding, acceptance, and use of advanced biofuels by promoting research, development and improvement of advanced biofuels technologies, production, marketing and delivery; and by promoting the sustainable development, cultivation and processing of advanced biofuels crops, and agricultural and forestry residues and wastes.” These slides are for public consumption and can be duplicated, replicated, modified, adapted, distributed, transmitted, and/or shared as seen fit by the reader. Please credit sources accordingly. If you wish to modify this document, just add your name under mine on the first slide. Note: Some slides contain additional information in notes section below

Energy: The Root of All Civilization 2. Why Bioalcohols? Concerted efforts from scientists, farmers, politicians, and grassroots organizations like Advanced Biofuels USA to understand and advocate for sustainability are ongoing. Most of us are seeking the promise of global security, the development of a sustainable workforce, and an endless supply of clean renewable energy. Converting biomass to biofuels for transportation fuel applications is currently one of the most active areas of investigative research in science and engineering. The following sections will offer an in-depth technical perspective of liquid fuels and demonstrate the overriding potential of bioalcohols to bridge transportation energy needs of modern society with the future of the human race. Energy: The Root of All Civilization 2. Why Bioalcohols? Blending Bridges to Sustainability 3. Leaping Barriers: Squeezing the Sun

The root of all civilization Section 1 Energy: The root of all civilization This section will connect energy to GDP and population size to illustrate the rise of human civilization and prosperity.

In the beginning there was…..biofuels? 1 EJ = 1018 J The post civil war exploitation of coal helped spawn the Industrial Age, while the subsequent incorporation of crude-oil and natural gas fossil resources helped spawn what has become a global economy. Is this pattern sustainable? Most believe the answer to this question is NO! Why?

In this section the ongoing energy crisis can be visualized in a series of graphs depicting the startling connection between: Energy Consumption GDP per capita (prosperity) Population growth Debt (deficit spending)

Almost a vertical slope!

And the United States population has been growing dramatically And the United States population has been growing dramatically. Projections indicate that the US could have a population over 400 million by 2050. To sustain such a state given our free market culture would require much more energy or a breakthrough in energy conservation technology (smart grid). chart by : http://perotcharts.com/2008/05/growth-of-us-population-1790-2050/

The fossil fuel dynasty has really just began, and yet it may already be more than half over. http://8020vision.com/2010/06/21/the-real-population-problem/

U.S. primary energy use by fuel (1980-2035) 1.0 × 1015 Btu 40% 1.0 × 1015 Btu equals a 1 quadrillion Btu (a quad). Most energy data is reported in Btu (British Thermal Units). U.S. Energy Information Administration (Washington, DC, June 2009) Projections: AEO2010 National Energy Modeling System

Breakdown of the U.S. liquid fuel market less than 3% biofuels 35 quadrillion Btu’s (37 EJ) of liquid energy annually ~ 95% of all liquids since 1958 have come from petroleum 1 63% of refined petroleum was delivered to market as motor gasoline for transportation2 less than 3% biofuels Energy Information Administration, Annual Energy Review 2008, Petroleum Consumption: Transportation Sector, 1949-2008. 2009, U.S. Department of Energy, Washington, D.C O’Donnell, M. Master’s Thesis, University of Texas at Austin, 2009

Global transportation energy consumption vs. GDP in 2006 A more specific look at transportation vs. prosperity. Many questions arise from this graph. For example, can the US sustain population growth, energy consumption, increasing debt, and a strong GDP growth?

World energy use approaches 500 quadrillion Btu’s, of which the US consumes approximately 1/5. graph from: http://environmentalresearchweb.org/blog/2009/07/high-debt-and-energy-return-on.html

Prosperity (GDP) correlates strongly with energy use Prosperity (GDP) correlates strongly with energy use. We know that China has benefited greatly from US prosperity, but most of its vast population doesn’t use much energy. Prosperity (GDP) correlates strongly with energy use. We know that China has benefited greatly from US prosperity, but most of its vast population doesn't use much energy. Liquid fuel energy for transportation is a key driver of energy expansion and China will need much more of it as they expand their transportation and industrial sectors. Knowing the technical difficulties we are currently experiencing, once again this pattern is visibly unsustainable and it also appears the US has much to lose.

Energy and Economic Interconnectedness This diagram depicts the feedback loops between an economy that is dependent on an aging oil industry. Energy and Economic Interconnectedness http://tclocal.org/images/failure-feedback.jpg

Summary It appears there exists a positive correlation between energy consumption, population growth rate, GDP, and the abstractions of expanding debt and monetary instability. So now what? We must now consider alternatives to the current trends of fossil fuel dependence and moves toward sustainability. The next section will discuss the biofuels option with an in-depth analysis of the bioalcohol paradigm.

Blending Bridges to Sustainability Why Bioalcohols? Blending Bridges to Sustainability Section 2

What are biofuels? In contrast to fossil fuels, biofuels…. Biofuels are any biologically derived solid, liquid, or gas that stores energy used in combustion applications. In contrast to fossil fuels, biofuels…. Are sustainable (1-100 yrs vs. 106-108 yrs) Can be carbon neutral or negative Have a more diversified, distributed means of production 4. Can be created as reagent grade molecules (pure)

Bioalcohol Biodiesel BIOFUEL TYPES Biogas Biocrude Biowaste Biogasoline (grassoline) Biogas Biodiesel BIOFUEL TYPES Biomass

Commercially available Under investigation and development Alternative Transportation Fuels Commercially available Methanol Natural Gas Propane Biodiesel Electricity Ethanol Hydrogen Under investigation and development Biobutanol Fischer-Tropsch (FT) diesel Gas to Liquids (GTL) Biogas Biomass to Liquids (BTL) Coal to Liquids (CTL) Hydrogenation-Derived Renewable Diesel (HDRD) P-Series (gasoline substitute) Source: The Energy Policy Act (EPAct) of 1992

adapted by: Drew Sowersby biofuels conversion biomass Historical reminder of expansive energy consumption in the US (1800-2000) and almost non-existent impact of biofuels. There is a very high mountain to climb for biofuels to replace fossil fuels. Before fossil fuels were available, humans relied almost 100% on biofuels (mostly wood for burning and feed for working animals such as horses, oxen, llamas, etc.) and human energy. adapted by: Drew Sowersby

Source: U.S. Department of Energy’s Energy Information Agency (EIA). Million Barrels per Day chart by: http://tclocal.org/images/eia-liquidfuels.jpg Source: U.S. Department of Energy’s Energy Information Agency (EIA).

Global biofuel supplies expected to increase dramatically less than 2% of total liquid consumption Million barrels day more than 90% of all cars use sugarcane ethanol BP p.l.c., Statistical Review, BP Energy Outlook 2030, London, January 2011

The evolution of biofuels is defined in terms of the carbon feedstock used for production 1st generation fuels corn-starch sugar from cane and beets soy for diesel 2nd generation – multi-component cellulose switchgrass miscanthus agriculture and food processing residues poplar trees 3rd generation – high quality cellulose microalgae macroalgae (seaweed) cyanobacteria 4th generation - sun fuels carbon dioxide + light + biocatalyst… CO2 impact factor (medium to high lignin content) (low to no lignin) net 0

Bioalcohols currently dominate commercially available biofuels biomass sugar feedstocks fermentation product recovery market 1 chemical Storage market 2 market 3 The Bioalcohol Paradigm

Biomass to Biofuels The process of biofuels production begins with the sun. Several crops with different properties can be grown in various climates and harvested as a feedstock for biofuels production. An array of processing and engineering techniques are used to convert or extract the biomass and produce energy for electricity, heat, and transportation. Notice that cellulosic ethanol, a second generation advanced biofuel, along with biodiesel represent the baseline targets for implementation into the liquid fuel chain. It is implied that many jobs will be created at each of these steps….and all are sustainable. http://www.vsjf.org/project-details/13/biomass-to-biofuels-resources

process generalization biomass bioalcohols This is a basic (although already extremely complex) process for creating 2nd generation biofuels from cellulosic biomass. Essentially, most bioalcohols are produced via a very similar pathway. In addition, a gasification process can be used instead of the pre-hydrolysis and hydrolyse steps before fermentation.

Most cellulosic material, like woods and grasses, contains lignin Lignocelluloses represent the most abundant source of bioenergy Glucose Treatment with cellulases and/or acids releases glucose monomers for fermentation Rubin, E. Nature, 2008, 454, 841-845.

But lignocellulosic feedstocks are not easily converted to sugar substrate and can introduce over 100 inhibitors into fermentation batches1 organic acids phenols aldehydes ketones CLASSES of inhibitors Liu, Z. L.; Slininger, P. J.; Gorsich, Appl Biochem. Biotechnol., 2005, 124, 451-460.

So far, Saccharomyces cerevisiae have demonstrated the ability to perform with a lignocellulosic feedstock. insulin lactic acid carotenoids alcohols carbon dioxide polymer precursors Advantages Are the most common microorganisms used for production of biofuels (primarily alcohols) Are eukaryotic Have simple nutrient requirements Are prime targets for bioengineering Convert glucose to ethanol with unusual efficiency (FERMENTATION) The yeast cell factory has been used by humans for over 8000 years to create a host of useful renewable products

Higher alcohol synthesis Ehrlich Pathway Glucose Pyruvate O2 Glycolysis (regulated and irreversible steps) CO2 + H2O respiration Fermentation amino acid synthesis CO2 + CH3CH2OH Standard fermentation in yeast Higher alcohol synthesis

BCAAs Branched-chain alcohols decarboxylation (step 2) NADH-dependent reduction (step 3) transamination (step 1) BAT1, BAT2 PDC1, PDC2, PDC3, PDC5, PDC6, ARO10, THI3 (KID1) ADH1, ADH2, ADH3, ADH4, ADH5, ADH6, SFA1, etc. 2MB 2MP 3MB (leucine, valine, isoleucine) 2-Keto acids Yeast cells naturally create C4 and C5 alcohols using fermentation enzymes Ehrlich Pathway branched-chain alcohol synthesis superior alcohol fuel surrogates Ketoaldehydes + CO2

Fermentation as a complex adaptive system Nitrogen Source Gases (CO2 and O2) Water Excess sugar Ionic Strength pH Inhibitors Viscosity Fluid Motion Temperature Biocatalyst Hypothetical Interaction Map

Isobutanol (2MP) is a viable platform molecule conventional motor gasoline isobutanol GEVO, Inc. Highlights High yield isobutanol yeast fermentation (105 g/L per batch) Conversion to hydrocarbons Carbon emissions reduction of 85% Competes with oil at $65 a barrel source: GEVO, Inc.

C4-C5 Alcohol Platform ButylFuel, LLC Highlights Case Study: After logging 10,000 miles butanol…. increased auto mileage by 9% reduced oxides of nitrogen by 37% reduced carbon monoxide to 0.01% reduced hydrocarbons by 95% first American company to commercialize butanol Case Study: “Production of Butyric Acid and Butanol from Biomass” Ramey D and Yang S-T, Phase II STTR Final Report for D.O.E. (2004)

C4-C5 alcohols have advantages compared to ethanol higher energy density lower vapor pressure lower air/fuel ratio less corrosive less hygroscopic higher gasoline blend ratios “drop-in” fuel compatible with gasoline engines, existing storage facilities, and distribution infrastructure Harvey, B. J.; Meylemans, H. A. J Chem Technol Biotechnol., 2011, 86, 2–9. Dürre, P. Biotechnol. J., 2007, 2, 1525-1534.

Selected bioalcohol and gasoline properties Fuel Cn Energy density (MJ/L) Boiling point (°C ) Solubility in water at 20°C (g/L) Vapor pressure at 20°C (mm Hg) Gasoline 4-12 33 38-204 negligable 275-475 Ethanol 2 21 78 miscible 59 2-methyl-1-propanol* 4 26 108 95 9 3-methyl-1-butanol 5 28 130 30 2-methyl-1-butanol 128 36 (at 30°C ) 3 Butanols have 4 carbons per molecule and pentanols have 5. miscible = fully mixed or homogeneous Cn = carbon atoms per molecule MJ/L = mega joules per liter mm/Hg = millimeters of mercury --information obtained from MSDSs, Sigma-Aldrich website, and NIST chemistry WebBook. * a.k.a. isobutanol ~ 1-butanol

Liquid Fuel Energy Densities MJ/L butanol/pentanol sweet spot? C4 alcohols such as 1-butanol (or isobutanol) have a lower knock-index, and burn more efficiently and cleanly than gasoline. C5 pentanols such as 3-methyl-1-butanol have not been thoroughly tested for combustion properties but are expected to behave similarly to 1-butanol. MJ/kg Source:Scott dial http://en.wikipedia.org/wiki/File:Energy_density.svg Adapted by Drew Sowersby

Right now fuel blends are showing up at pumps across the U.S. BRIDGE FUELS E10 Up to 10% ethanol to replace MTBE E15 - E85 contains 15% to 85% ethanol requires post 2001 or Flexfuel engine technology B20 contains 20% biodiesel / 80% diesel made commercially from soybeans How long until we see C4 and C5 advanced alcohols at the pump?

Section 3 Leaping Barriers: Squeezing the Sun

The Obstacle Course It would be irresponsible to assume that human energy needs will be fulfilled in a timely fashion. The transition to sustainable energy will likely be a long arduous process. Moore’s Curse and the Great Energy Delusion (The American Magazine, November 19, 2008) “There is one thing all energy transitions have in common: they are prolonged affairs that take decades to accomplish, and the greater the scale of prevailing uses and conversions the longer the substitutions will take. The second part of this statement seems to be a truism but it is ignored as often as the first part: otherwise we would not have all those unrealized predicted milestones for new energy sources.” - Vaclav Smil-Distinguished Professor at the University of Manitoba.

Sheer size required for economic growth Geographic distribution Technical Barriers GOAL Sheer size required for economic growth Geographic distribution START Supply continuity Low crop energy density Kerr, R. Science, 2010, 329, 780-781

The Bright Side The sun delivers about 1000 W/m2 of power to Earth’s surface. 1000 Wh = 1 kWh = 3.6 mega Joules (MJ) peak sun hour = 1 kWh peak sun hours per day based on geo location http://pvcdrom.pveducation.org/SUNLIGHT/AVG.HTM

U.S. example? ≈ 5.20 × 1016 MJ/year ≈ 4.90 × 1019 Btu/year ≈ 4.00 peak sun hours avg./day1 1 peak sun hour = 3.6 MJ 14.4 MJ/(m2)day × 365 days × 9.83 × 1012 m2 ≈ 5.20 × 1016 MJ/year 1 MJ = 994.78 Btu ≈ 4.90 × 1019 Btu/year U.S. example? US land area this is roughly 500X the current amount of US energy usage 1. Solar Radiation Data Manual for Flat-Plate and Concentrating Collectors National Renewable Energy Laboratory (NREL), 2006

Earth? ≈ 1.35 × 1018 MJ/year ≈ 1.28 × 1021 Btu/year ≈ 2.00-3.00 peak sun hours/day 7.2 MJ/(m2)day × 365 days × 5.14 × 1014 m2 ≈ 1.35 × 1018 MJ/year ≈ 1.28 × 1021 Btu/year

“Using detailed land analysis, Illinois researchers have found that biofuel crops cultivated on available land could produce up to half of the world's current fuel consumption – without affecting food crops or pastureland. Adding LIHD (low input high density) crops grown on marginal grassland to the marginal cropland estimate from earlier scenarios nearly doubled the estimated land area to 1,107 million hectares globally, even after subtracting possible pasture land – an area that would produce 26 to 56 percent of the world's current liquid fuel consumption.” -- http://cee.illinois.edu/cai_biofuel_land Published in the journal Environmental Science and Technology, the study led by civil and environmental engineering professor Ximing Cai identified land around the globe available to produce grass crops for biofuels, with minimal impact on agriculture or the environment.

What will the next transition be? Paradigm Shift standard fermentation to advanced fermentation 2nd generation biofuels 1st generation NON-FOOD crops and waste/residues FOOD crops CO2 and the SUN

Taking Us from the Present to the Future Many companies are engaged in making these transitions happen. See a list of more than 400 companies in the Resources section on the Advanced Biofuels USA web site: http://advancedbiofuelsusa.info/resources/companies-involved-with-advanced-biofuels Find out more at www.AdvancedBiofuelsUSA.org