Harvesting Algae to Form a Neutraceutical, Specifically Creating a Functional Food

1. Harvesting Algae to Form a Neutraceutical, Specifically Creating a Functional Food Team Alpha Travis Dallas Eric Graves Joaquin Martinez Chris McNinch Ramune Otterson Meskyte Charu Saini Crypthecodinium Cohnii

2. Beer Still Bottom Water Glucanex Biomass Water Hexane Glucose Water Antifoam Vent Sterile air Fermentation Dissolved Air Floatation CO2 Lysing Centrifuge Hexane Extraction Algae Salt CIP System Steam Cooling Water Air CO2 Sodium methoxide Citric Acid Water Refrigerant Interesterification Emulsification Pasteurization Scraped Surface Heat Exchanger Gum Water Citric Acid Acid Activated Clay Degumming Centrifuge Bleaching Removal FFA Deodorization Water Crude SBO Packaging Steam Sea Salt Hardfat SBO Lecithin Sodium methoxide Diacetyl Steam Citric Acid Mono/Diglycerides Citric Acid Citric Acid Water B-Carotenes To Wholesaler Enzyme Acid Activate Clay Sorbic Acid Water Ergocaliferols NaOH

3. Key Points • Plant Layout • Controls • Sanitation Design • Calculations • Economics • Changes • Future Endeavors • PFD • Report

4. Gate Wind Direction D A B Fencing C E F • Storage/Boilers • Fermentation • Offices/Labs/Miscellaneous Space • Water Cooling Tower • SBO Refining • Margarine Processing General Plant Layout Alpha Algae Plant 1928 SE Otis Rd Cedar Rapids, IA 52401 Otis Rd Cargill

5. Fermentation Layout 115 ft Cooling Tower Ethanol Bottoms Salt Boiler Glucose Storage Electrical Room 35 ft Glucanex Antifoam Seed 1 Fermenter 1 RXN 1 Harvest Tank S1 RXN 2 Seed 2 Fermenter 2 S2 DAF 85 ft Absorption Mix Tank Lipid Storage Fermenter 3 Lysed Tank Seed 3 S3 Fermenter 4 Centrifuge 1 Centrifuge 2 Seed 4 S4 C3 C4 Dryer Silo

6. Refining Layout SoftWaterStorage CrudeSBOStorage DegumSBOStorage Surge Tank Mixers WaterSoftener Batch Rx Cent-5 Vacuum Mixer Surge Tank RefinedSBOStorage VacuumVessel ElectricalRoom Mixers InteresterSBOStorage InteresterificationTank HardfatSBOStorage Cent-7 Cent-6 CA ProcessingTank 72ft x 34ft

7. 50’ x 66’

8. Margarine Plant Mezzanine Layout Ingredient Storage Tank Area Electrical Room = 4 feet

9. QIC QIC QIC 1 QIC 3 2 11 TIC 4 10 5 TIC 7 4a QIC QIC 9 15 14 QIC FIC 8 TIC TIC 12 PIC PIC PIC PIC FIC FIC Fermentor Controls TK-2 TK-1 TK-3 TK-4 TK-5 P- 2 P- 10 Glucose Salt P- 1 P- 5 P- 6 Ethanol Bottoms STEX-1 Filter 3 MT-1 Direct Steam Injection Sterile Glucanex P- 11 P- 3 Water Sterile Anti-foam Water Steam Vented Air Filter 2 13 Vent 10% Seed Fermenter Harvest Tank Main Fermenter 250 mL Shake Flasks 1000 L Seed Fermenter 10 L Seed Fermenter To DAF P- 8 P- 7 P- 9 Sparged Steam Cooling Water Sterile Air Compressor 1 Sparged Steam Air Cooling Water Filter Filter 1 1 Sterile Air Compressor 1 Air

10. QIC PIC QIC QIC QIC Visc 7 QIC QIC TIC PIC FIC TIC Fermentor Controls Sterile Glucanex Filter 3 Sterile Anti-foam 11 P- 6 10 P- 3 Vented Air Filter 2 13 Foam Main Fermenter Turbidity P- 8 14 9 Cooling Water Sparged Steam 8 Filter 1 Compressor 1 Sterile Air Air

11. Dissolved Air Flotation Controls 20 Vented Air DAF Tk-7 1 Fermented Algae Tk-6 16 Cv-23 Cv-25 P11 FIC FIC Rxn tank B 22 23 FIC P12 Cv-20 Conc. Algae 15 18 P-01 19 FIC 2 FIC Tk-7 Cv-22 Cv-26 17 Rxn tank A FIC Fermented Algae 21 P10 Sterilized Air Water 45 PIC Water PIC 46 12 18 18 Water to Cargill treatment Cv-21 Cv-24 Sterilized Air Sterilize Air Sterilized Air

12. CO2 Lysing Controls Cv-30 Conc. Algae CO 2 Compressor 2 27 23 Vented CO2 3 Stir tank Filter 4 Lysed Algae Tower Tk- 9 Baffle tray Tower Tk- 8 24 Cv-29 PIC Sterilized CO2 Cv-30 FIC FIC Cv-28 25 26 29 CO2 Absorbed Algae Lysed Algae & CO2 Throttle Valve P14 Lysed Algae TIC 28 Cv-31 48 P13 47 FIC Cv-27 Steam in & out

13. Main Centrifuge Controls Algae Oil Holding tank TK-20 Lysed Algae 29 Algae Wet Biomass Algae Oil 30 31 Disc Centrifuge Cv-32 FIC 35 Algae Oil P15

14. Separation Process Controls Cv-36 FIC Wet Biomass Tank Tk-12 40 Wet Biomass Wet Biomass 41 P19 Biomass Cv-37 FIC Dynamic Cross Flow Filtration 42 39 Water P21

15. Dryer Process Controls Exhauster Purge Air seal dust valve Fume controller 37 42 PIC Recycle Filter Rotary Dryer 49 43 Dry Biomass Steam in 50 Steam out Conveyor Belt TIC Collection Hopper Shipping container Dried Algae Distributor Conveyor Belt

16. Degumming Controls TK-53 4% NaOH Storage TK-54 10% Enzyme Refrigerated Storage PIC N2 Gas TK-52 45% Citric Acid Storage P-54 P-53 TC TK- 51 Crude SBO Storage PIC Batch Reactor FIC N2 Gas TK-55 Surge Tank QIC pHC LIC P-52 QIC HX-5 HX-6 PIC P-56 LIC P-51 P-55 TIC N2 Gas LIC TIC CLR-1 LLO HLO - Gums TIC FIC TK-56 Degummed SBO Storage Cent-5

17. Refining Controls TK-58 Acid Activated Clay Tank TK-59 Water Storage PIC VP-1 TK-57 50% Citric Acid Storage PIC VP-2 Vacuum Vessel FIC N2 Gas Vacuum Mixer PIC TK-56 Degummed SBO Storage RP-1 CLR-2 P-60 P-58 FIC TIC FIC TIC P-62 HX-11 LIC HX-9 Filter HX-8 P-57 P-59 P-61 PIC MX-1 MX-1 FIC N2 Gas Steam TIC TK-60 Surge Tank To MX-101 LIC P-64 PIC N2 Gas LIC P-63 FIC HX-10 CLR-3 TK-61 Refined SBO Storage To Vacuum Mixer

18. PIC Interesterification Controls N2 Gas TK-67 Soft Water Storage Water Softener TK-62 SBO Hardfat Storage LIC QIC FIC P-71 P-65 PIC HX-16 N2 Gas FIC TIC TK-63 SBO Hardfat Heating Tank TK-64 CH3ONa Storage TK-65 20% Citric Acid Storage FIC MX-3 MX-4 P-68 P-69 TIC TC PIC P-66 HX-14 LIC QIC FIC N2 Gas P-72 P-73 PIC TIC HLO HLO N2 Gas TC TK-66 Mixing Tank LIC TK-61 Refined SBO Storage QIC Cent-6 Cent-7 PIC TK-68 SBO Storage N2 Gas P-67 HX-15 TC TIC P-70 CLR-4 TIC FIC FIC Filter

19. Margarine Controls 1 QIC Algae Oil Storage TK-20 P-30 CIP CIP CIP QIC Refined SBO Storage Warm Water TK-21 P-31 CIP QIC LecithinStorage TIC Oil Oil Phase Prep. Tank 1 TK-22 QIC P-35 Beta CaroteneStorage TK-23 TK-31 QIC QIC Diacetyl Storage TK-24 CIP QIC P-34 Sorbic Acid Storage E TK-25 Oil Oil Phase Prep. Tank 1 CIP QIC Mono- di glycerides Storage TK-26 TK-32 QIC QIC Aqueous Phase Prep. Ergo- califerol Storage TK-27 P-33 QIC TK-30 TIC Water Storage P-35 TK-28 Sea Salt Storage QIC Warm Water P-32 TK-29 Pasteurization Warm Water

20. Margarine Controls 2 P-36 Plate HX TIC CIP CIP High Pressure Pump PIC PIC Pin Rotor Machine Scraped Surface Heat Exchanger P-37 VIC TIC Tub Filling and Packaging Ammonia Compressor

21. CIP Controls Water CIP Returning Liquid CIP Supplying Liquid P-42 FIC Strainer TIC LIC LIC LIC LIC FIC LIC LIC LIC LIC P P CS HX-41 TK-41 Acidic Solution TK-42 Alkaline Solution TK-43 Reclaim Water TK-44 Fresh Water P-41 Alkaline Storage Acidic Storage Drain

22. Sanitary Design Equipment Design Cleanable to microbial level Made of compatible materials Accessible for inspection, maintenance and cleaning No liquid collection and no niches Equipment must perform as designed Hygienic compatibility with other plant systems Validated cleaning procedures

23. Sanitary Design (cont) Facility Design Physical separation of distinct hygienic zones Material flow and personnel movement control Prevent water accumulation inside building materials Temperature and humidity control Air flow and air quality control Building envelope sanitary conditions Interior spatial design that enables cleaning Sanitation integrated into facility design

24. Calculations – Fermentation Process Fermentation Time Requirement (hours/batch) = Lag Phase + Growth Phase + Stationary Phase + Maintenance = 0 + [ln(27.7 g/L)-ln(2 g/L)] / 0.03747 hr-1 + 20 + 12 = 102.14 hours Algae Production (tons/batch) = Final Concentration (g/L) x Broth Volume (L) / (1000 g/kg) x (2.2046lb/kg) / (2000lb/ton) = 27.7 x 79,500 / 1000 * 2.2045 / 2000 = 2.427 tons/batch Annual Amount of Algae Produced (tons) = Oil needed (tons) / 0.2 (g lipids / g dw cells) = 133 / 0.2 = 665.0 tons Annual Amount of Batches Required = 665.0 tons / (2.427 tons/batch) = 274.0 batches Plant Operating Time (hours/year) = Time per batch (hours) * Batches per year / Number of fermentors = 102.14 hours * 274.0 / 4 = 6,996.6 ~ 7,000 hours/year

25. Plant Economics Total Project Capital Cost 37,328,232.77 USD Operating Costs18,962,032.85 USD/Year Raw Materials 6,031,280.00 USD/Year Utilities 3,532,157.83 USD/Year Maintenance 2,030,000.00 USD/Year Operating Labor Costs 2,828,000.00 USD/Year Operating Charges 707,000.00 USD/Year Plant Overhead 2,429,000.00 USD/Year G and A Costs 1,404,594.96 USD/Year Total Product Sales 25,963,238.43 USD/Year Payback Period 5.7 Years Tax Rate is 40% Straight Line Depreciation over 10 years 2,986,258.62 USD/Year

26. Changes

27. Separation Process PFD Wet Biomass Tank Tk-12 40 Wet Biomass Wet Biomass 41 P19 Biomass Dynamic Cross Flow Filtration 42 39 Water P21

28. Dryer Process PFD Exhauster Purge Air seal dust valve Fume controller 37 42 Recycle Filter Rotary Dryer 49 43 Dry Biomass Steam in 50 Steam out Conveyor Belt Collection Hopper Shipping container Dried Algae Distributor Conveyor Belt

29. Degumming (Changes) TK-53 4% NaOH Storage TK-54 10% Enzyme Refrigerated Storage N2 Gas TK-52 45% Citric Acid Storage 131 P-53 P-54 N2 Gas TK- 51 Crude SBO Storage 138 139 143 Batch Reactor TK-55 Surge Tank P-52 HX-5 HX-6 136 132 133 137 142 144 N2 Gas TK-56 Degummed SBO Storage P-51 P-55 P-56 135 147 145 153 134 146 141 140 CLR-1 LLO HLO - Gums 148 149 150 152 151 Cent-5

30. N2 Gas Interesterification (Changes) 193 TK-67 Soft Water Storage Water Softener TK-62 SBO Hardfat Storage P-71 P-65 214 N2 Gas 194 217 HX-16 195 218 TK-63 SBO Hardfat Heating Tank TK-64 CH3ONa Storage TK-65 20% Citric Acid Storage 215 216 196 197 MX-3 MX-4 P-68 P-69 P-66 HX-14 N2 Gas 219 222 198 199 209 210 206 P-72 P-73 N2 Gas HLO 220 221 224 223 HLO 201 192 200 TK-66 Mixing Tank TK-61 Refined SBO Storage Cent-6 Cent-7 N2 Gas 207 TK-68 SBO Storage 208 228 P-67 HX-15 202 203 P-70 213 CLR-4 211 225 205 212 204 Filter 227 226

31. Margarine PFD 2 (Changes) 82 81 P-36 80 Plate HX 83 CIP CIP High Pressure Pump Pin Rotor Machine Scraped Surface Heat Exchanger 84 87 88 P-37 86 89 85 Packaging 90 Ammonia Compressor

32. Calculation - Margarine Length of Pasteurization Piping needed Volumetric Flow rate/(Area of pipe) = velocity of fluid .0155 m3/min/(π * .022252) = 9.966 m/min 9.966 m/min * 10 min = 99.66 m needed for pasteurization time 99.66 m (3.28 ft / 1 m) = 326.885 ft Or 330 ft of insulated pipe needed for pasteurization

33. CIP PFD Water CIP Supplying Liquid CIP Returning Liquid 214 201 203 205 213 P-42 Strainer 216 217 215 207 210 211 212 TK-41 Acidic Solution TK-42 Alkaline Solution TK-43 Reclaim Water TK-44 Fresh Water P P HX-41 209 218 206 202 204 208 P-41 Alkaline Storage Acidic Storage Drain

34. Future Endeavors • Within next 5 years take over 2% of the market • Need to add 3 fermentors, remainder of plant can be operated at higher capacity • Degumming and Deacidifiction byproducts streams • Lecithin profitable byproduct • FFA high in protein into animal feed • Margarine Flavoring – cinnamon, garlic, honey • Strictly selling DHA as vitamin supplement at GNC

35. Margarine Nutritional Label

36. Summary Plant Layout Controls Sanitation Design Calculations Economics Changes Future Endeavors PFD Report

37. Questions?

38. Specification on Rotary Dryer (Heyl & Patterson) • Dryer Type = Direct Rotary • Process Type = Batch Mode • Dryer Size = 90" Diameter x 60 ft. O/A Length • Air Flow Rate = 5430 lb/hr • Steam Heat Exchanger Duty = 271,065 BTU/hr Max. • Dryer Duty = 101,650 BTU/hr Max. • Dryer Inlet/Outlet Air Temperature = 268/190 Deg F • Dryer Evaporative Load = 3829 lb/hr • System Price = $400,000-$660,000 USD

39. Calculations – Fermentation Process Steam Sparged into Fermentor to Sterilize, 60 to 121°C (kg steam) (Steam at 145°C and 3 barg) mbroth (kg)*cp(kJ/kg*K)*ΔT (K) = msteam(kg)*Hvap (kJ/kg) + msteam (kg)*cp (kJ/kg*K)*ΔT (K) 17,300,000 kJ = msteam *[2,163.47 (kJ/kg) + 0.5*4.187*(144-120)+0.5*4.187*(144-60)] msteam = 7,240 kg steam Steam Used to Heat Mix Water from 17 to 60°C (kg steam) (Steam at 145°C and 3 barg) mbroth (kg)*cp(kJ/kg*K)*ΔT (K) = msteam(kg)*Hvap (kJ/kg) + msteam (kg)*cp (kJ/kg*K)*ΔT (K) 9,873,628 kJ = msteam *[2,163.47 (kJ/kg) + 0.5*4.187*(144-17)+0.5*4.187*(144-60)] msteam = 3,719.7 kg steam Heat Evolved from Cell Growth (kcal) = 0.12 * Oxygen Uptake Rate (mmol/(h)) * Fermentation Time (h) = 0.12 * 295.35 mmol/h * 70.14 h = 2,485.88 kcal

40. Calculations - Separations Centrifuge #2 Mass Balance: ΣMass = Mass in – Mass out = 0 0 = 22,902.52 (kg/batch of lysed algae) – 6,168.82 (kg/batch of algae) – 16,733.71 9 (kg/batch of waste water) Energy Balance: Q = MCp(∆T) + Qin Q = assumption of no temperature change , Qin HP supplied to centrifuge Q = 100HP x 2545 = 244,500 BTU/ Hr Centrifuge #3 Mass Balance: ΣMass = Mass in – Mass out = 0 Energy Balance: Q = MCp(∆T) + Qin Q = assumption of no temperature change , Qin HP supplied to centrifuge Q = ? HP x 2545 = 381, 845 BTU/ Hr

41. Calculations - Separations Co 2 absorption and Lysing Mass Balance: ΣMass = Mass in – Mass out = 0 0 = 86,663 (kg/batch of algae) + 1000 (kg/hr of air) – 63,295 (kg/batch waste water) -23,368 (kg/batch of conc. algae) - 1000 (kg/hr of air) Energy Balance: Q = MCp(∆T) + Qin Q = throttling process is adiabatic, so Qin is the steam jacket to tank Q = 12,700.6 kg/hr of steam Centrifuge #1 Mass Balance: ΣMass = Mass in – Mass out = 0 0 = 23,368 (kg/batch of conc. Algae) - 466 (kg/batch of lipids) - 22, 902 (kg/batch of lysed algae) Energy Balance: Q = MCp(∆T) + Qin Q = assumption of no temperature change , Qin HP supplied to centrifuge Q = 150 HP x 2545 = 381, 845 BTU/ Hr

42. Calculations - Refining Vacuum Vessel Sparging Steam flow design Tray1- 715.59/6/3785.412*0.092/1000*0.001 = 0.42 gal/hr Tray2- 715.59/6/3785.412*0.092/1000*0.007 = 2.91 gal/hr Tray3- 715.59/6/3785.412*0.092/1000*0.015 = 6.23 gal/hr Tray4- 715.59/6/3785.412*0.092/1000*0.015 = 6.23 gal/hr HX-11 Oil into the Vacuum Vessel Qoil = mcp(∆T) Qoil = 921.33*0.55*(500-284) Qoil = 109454 BTU/hr QPoil = mcp(∆T) T = 617- (109454/2900/.79) T = 569.2°F ∆Tlm = [(T1 – t2) – (T2 – t1)]/ln[(T1 – t2)/ (T2 – t1)] ∆Tlm = [(617-500) – (569.2-284)]/ln[(617-500)/(569.2-284)] ∆Tlm = 188.8°F Q = UA ∆Tlm A = 109454/60/188.8 A = 9.68 ft2