Photosynthetic Bio-Manufacturing in Algae Food and Fuel for the 21st Century Designing Photosynthetic Microorganisms Stephen Mayfield The California

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1 Photosynthetic Bio-Manufacturing in Algae Food and Fuel for the 21st Century Designing Photosynthetic Microorganisms Stephen Mayfield The California Center for Algae Biotechnology University of California San Diego

2 What is bio-manufacturing? Is the use of living systems and organisms to produce useful products

3 Agriculture is also bio-manufacturing Algae is Agriculture

4 Why Food and Fuel together? Both are Chemical Energy: The energy in a substance that can be released by a chemical reaction to do work!

Food & Fuel Two Sides of the Same Coin Chemical Energy Food

calories of food per day 1 gallon of gas = 31,500 calories 1

8 gallons EtOH Average American consumes 40,000 calories of

5 Food & Fuel Two Sides of the Same Coin Chemical Energy Food Energy Fuel Energy Average American consumes calories of food per day 1 gallon of gas = 31,500 calories 1 bushel of corn = 84,000 calories 1 bushel of corn = 2.8 gallons EtOH Average American consumes 40,000 calories of fuel per day % corn crop was turned into 14 billion gallons of fuel Ethanol

6 We convert fuel into food through Industrial Ag The Green Revolution brought Enormous Yield Increases over the last 50 years

7 Thousands of barrels/day Oil Production Over the Last 10,000 Years Time (years)

8 Thousands of barrels/day Millions of people World Population and Energy Production over the last 10,000 years Time (years)

9 The food price index can be overlaid On the oil price index

10 As people gain wealth they tend to eat more animal protein

11 This is directly related to world wide decline in fish populations

https://www.nytimes.com/2017/02/24/business/energy-environment/deforestation-brazil-bolivia-southamerica.

12 And to increased deforestation for agriculture production Google: Cargill Deforestation New York Times

13 Why algae? What are algae? Algae are a new photosynthetic biomanufacturing Why use algae? platform 1. Grow fast What can we make from algae? 2. Consume CO2 3. Thrive in wastewater and salt water 4. High biofuel yields 5. Doesn t compete with food production Sunlight Fuel Water Nutrients Plastics Animal Feed Carbon dioxide Inputs Conversion Engine Pharmaceuticals Outputs

14 Why algae as a platform for Photosynthetic Bio-manufacturing Scalable Productive Sustainable Programmable Edible 1.2 Trillion gallon of petroleum are consumed in the world every year 700 billion pounds of protein are consumed in the world every year

15 Processing Green Crude to Fuel Grow Harvest Extract Concentrate Crude Algae Oil can go directly to existing oil refineries to produce drop-in transportation fuels

Fungible fuels have been made from algae oil

algae-based renewable gasoline or diesel 2009

16 Fungible fuels have been made from algae oil Algaeus, hybrid vehicle crossed the country on algae-based renewable gasoline or diesel Sapphire Many air flights using algae jetfuel blends 2009 Sapphire US Navy amphibious vehicle run on algae Biodiesel blend Solazyme

17 Improving the Economics of Algal Biofuels Cost Per Gallon (in 2009 dollars) Bio-prospecting Engineering Breeding-selection co-products petroleum ppg Year

18 Large scale utilization requires low cost Low cost requires large scale utilization

19 Algae bio-manufacturing can produce a wide spectrum of products lipids Low Cost High Volume carbs protein s High Cost Low Volume

20 2 0 Strategy for the production of high value products in algae Identify the product Enable algae to produce that product Grow algae low cost and scalable High value products Product Processing Harvest algae

Colostrum is produced in all mammals contains highly active orally available proteins 33 detailed studies of Colostrum MAA and

human adults 2,754 total mammalian subjects Mammary Associated Amyloid (MAA) Safety demonstrated 875 human subjects in 18

demonstrated Osteopontin (OPN) 318 human subjects in 7 independent studies A total of 1,772 days of study 100% positive

21 Colostrum is produced in all mammals contains highly active orally available proteins 33 detailed studies of Colostrum MAA and OPN are major components of colostrum from pigs, sheep, cows, and humans Tested rats, mice, pigs, calves, human babies, and human adults 2,754 total mammalian subjects Mammary Associated Amyloid (MAA) Safety demonstrated 875 human subjects in 18 independent studies A total of 23,178 days of colostrum administration ZERO negative health effects were reported Efficacy demonstrated Osteopontin (OPN) 318 human subjects in 7 independent studies A total of 1,772 days of study 100% positive results in reducing or eliminating the symptoms of gastrointestinal disease I never fight evolution Biology had millions of years to get it right

Animal trials show MAA-algae reduces symptoms of clinical diarrhea 100% Cases of clinical diarrhea 50 Diarrheal days Piglets were challenged with pathogenic bacteria Piglets fed MAA-algae had a 62%

22 Animal trials show MAA-algae reduces symptoms of clinical diarrhea 100% Cases of clinical diarrhea 50 Diarrheal days Piglets were challenged with pathogenic bacteria Piglets fed MAA-algae had a 62% lower rate of diarrhea and a 48% fewer diarrhea days 80% 40 60% 30 40% 20 20% 10 Prevention and reduction of diarrhea could: 0% No treatment MAA algae Triton's product 0 No treatment MAA algae Triton's product Reduce the 5 million childhood deaths every year Replace antibiotics in both human and animal health MAA-algae protects post-weaning piglets from bacterial diarrhea

23 Petroleum is fossil algae oil: That means anything we can make from petroleum, we can make from algae Triacylglyceride + chemistry = polyhydroxyl FA or polyols

24 Polyols are the precursors to polyurethane Sustainable Surfboards Algae Oil-based Polyol TDI Toluene Diisocyanate + Polyurethane Polyol n

25 We also made renewable sandals

26 If you can make Rainbow colored algae You can make anything!

27 New Technologies must be proven to be safe actual trials In discussion with the US-EPA we designed the first outdoor GE algae field trial at UC San Diego

soil berm lined with mesh-reinforced, puncture

28 Construction of ponds for outdoor GM trial Secondary containment features of the field test: 8 sand and soil berm lined with mesh-reinforced, puncture resistant, UV-resistant pond liner, covered by sand. A C B

29 Obtained US - EPA Approval August 1, 2013 UCSD and Sapphire Energy submitted a Consolidated Toxic Substances Control Act (TSCA) Environmental Release Application (TERA) Which was approved on September 25, 2013

30 Measuring phenotype of wt and GE algae Both the GFP and modified lipid phenotype were observed throughout the test 60 days

31 Measuring dispersal of GM strain

distance North (meters) -10 0 10 20 30 40

> Days to invasion 59 34 > 59 21 32 26 42

32 distance North (meters) Measuring complexity of species in traps > Days to invasion > distance East (meters)

33 No effect of GE or wt on overall algal biomass

34 No effect of GM or wt on overall algal complexity

35 What did we learn from the outdoor GE algae trial Heterologous genes and phenotype are stabile in outdoor cultivation 50 days GE algae did not grow as well as wt outdoors Wt or GE algae can disperse took 50 days to travel 50 meters Many other algae showed up in traps immediately no large ponds near by and no significant rain during trial GE algae can reproduce in local water but do not over take native strains or reduce complexity

36 It took 20 months - four journals - and over 18 rounds of review

37 What did people say Almost everyone got it If we are going to produce food, feed, and fuel from algae, which the world needs, it will likely require genetic engineering Some Did not Many algae have horizontal gene transfer, so that makes any genetic engineering extremely risky With climate change in complete chaos this is no time to experiment with algae

38 Where do we go form here? Synthetic animal protein made from algae would be a game changer for the planet Algae is 100 times more efficient then cows at making high quality protein!

Teacher Background Information Making Fuel from Algae Lesson Plan

Teacher Background Information Making Fuel from Algae Lesson Plan 1. Fossil Fuels Fossil fuels are a finite resource including coal, oil, and natural gas. These were produced from materials such as algae