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

3 Key Points 1.Plant Layout 2.Controls 3.Sanitation Design 4.Calculations 5.Economics 6.Changes 7.Future Endeavors 8.PFD 9.Report

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

5 S2 Seed 1 S1 S4 S3 Seed 2 Seed 3 Seed 4 Mix Tank Fermenter 1 Fermenter 2 Fermenter 3 Fermenter 4 Harvest Tank DAF RXN 1 RXN 2 Absorption Lysed Tank Lipid Storage Centrifuge 1Centrifuge 2 C3C4 Dryer Glucose Storage Ethanol Bottom s GlucanexAntifoam Salt Boiler Cooling Tower Electrical Room 85 ft 35 ft 115 ft Fermentation Layout Silo

6 Hardfat SBO Storage Crude SBO Storage Water Softener Soft Water Storage Batch Rx Degum SBO Storage Mixers Cent-5 Vacuum Mixer Vacuum Vessel Refined SBO Storage Electrical Room Processing Tank Interester SBO Storage Interesterification Tank CACA Cent-6Cent-7 Mixers Surge Tank Surge Tank Refining Layout 72ft x 34ft

7 50’ x 66’

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

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

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

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

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

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

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

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

16 TK- 51 Crude SBO Storage P-51 N 2 Gas TK-52 45% Citric Acid Storage P-52 P-53 P-54 TK-53 4% NaOH Storage TK-54 10% Enzyme Refrigerated Storage N 2 Gas HLO - Gums Batch Reactor P-55 P-56 TK-55 Surge Tank N 2 Gas TK-56 Degummed SBO Storage HX-5 CLR-1 LLO HX-6 Cent-5 TIC FIC PIC TIC pHC QIC LIC TC FIC LIC Degumming Controls

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

18 N 2 Gas TK-61 Refined SBO Storage N 2 Gas TK-62 SBO Hardfat Storage N 2 Gas TK-63 SBO Hardfat Heating Tank P-66 P-67 HX-14 HX-15 P-68 TK-64 CH 3 ONa Storage TK-66 Mixing Tank P-69 P-70 Filter TK-67 Soft Water Storage HX-16 HLO P-71 P-72P-73 HLO Cent-7Cent-6 TK-65 20% Citric Acid Storage N 2 Gas TK-68 SBO Storage CLR-4 MX-3MX-4 Water Softener P-65 TIC PIC FIC TIC FIC QIC FIC LIC QIC TC FIC LIC TC N 2 Gas PIC FIC QIC LIC Interesterification Controls

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

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

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

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)] / hr = 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 * / 2000 = tons/batch Annual Amount of Algae Produced (tons) = Oil needed (tons) / 0.2 (g lipids / g dw cells) = 133 / 0.2 = 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 = hours * / 4 =6,996.6 ~ 7,000 hours/year

25 Plant Economics Total Project Capital Cost 37,328, USD Operating Costs18,962, USD/Year  Raw Materials 6,031, USD/Year  Utilities3,532, USD/Year  Maintenance2,030, USD/Year  Operating Labor Costs2,828, USD/Year  Operating Charges 707, USD/Year  Plant Overhead2,429, USD/Year  G and A Costs1,404, USD/Year Total Product Sales 25,963, USD/Year Payback Period 5.7 Years  Tax Rate is 40%  Straight Line Depreciation over 10 years  2,986, USD/Year

26 Changes

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

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

29 TK- 51 Crude SBO Storage P-51 N 2 Gas TK-52 45% Citric Acid Storage P-52 P-53P-54 TK-53 4% NaOH Storage TK-54 10% Enzyme Refrigerated Storage N 2 Gas HLO - Gums Batch Reactor P-55P-56 TK-55 Surge Tank N 2 Gas TK-56 Degummed SBO Storage HX-5 CLR-1 LLO 150 HX-6 Cent Degumming (Changes)

30 N 2 Gas TK-61 Refined SBO Storage N 2 Gas TK-62 SBO Hardfat Storage N 2 Gas TK-63 SBO Hardfat Heating Tank P-66 P-67 HX-14 HX-15 P-68 TK-64 CH 3 ONa Storage TK-66 Mixing Tank P-69 P-70 Filter TK-67 Soft Water Storage HX-16 HLO P-71 P-72P-73 HLO Cent-7Cent-6 TK-65 20% Citric Acid Storage N 2 Gas TK-68 SBO Storage CLR-4 MX-3MX-4 Water Softener P N 2 Gas 206 Interesterification (Changes)

31 High Pressure Pump Ammonia Compressor Pin Rotor Machine Scraped Surface Heat Exchanger Packaging P-37 CIP CIP Margarine PFD 2 (Changes) Plate HX 8182 P-36

32 Calculation - Margarine Length of Pasteurization Piping needed Volumetric Flow rate/(Area of pipe) = velocity of fluid.0155 m 3 /min/(π * ) = m/min m/min * 10 min = m needed for pasteurization time m (3.28 ft / 1 m) = ft Or 330 ft of insulated pipe needed for pasteurization

33 TK-41 Acidic Solution TK-42 Alkaline Solution TK-43 Reclaim Water TK-44 Fresh Water Alkaline Storage Acidic Storage CIP Returning Liquid Water HX-41 CIP Supplying Liquid Drain P-41 P-42 PP Strainer CIP PFD

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 1.Plant Layout 2.Controls 3.Sanitation Design 4.Calculations 5.Economics 6.Changes 7.Future Endeavors 8.PFD 9.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) m broth (kg)*c p (kJ/kg*K)*ΔT (K) = m steam (kg)*H vap ( kJ/kg) + m steam (kg)*c p (kJ/kg*K)*ΔT (K) 17,300,000 kJ = m steam *[2, (kJ/kg) + 0.5*4.187*( )+0.5*4.187*(144-60)] m steam = 7,240 kg steam Steam Used to Heat Mix Water from 17 to 60°C (kg steam) (Steam at 145°C and 3 barg) m broth (kg)*c p (kJ/kg*K)*ΔT (K) = m steam (kg)*H vap ( kJ/kg) + m steam (kg)*c p (kJ/kg*K)*ΔT (K) 9,873,628 kJ = m steam *[2, (kJ/kg) + 0.5*4.187*(144-17)+0.5*4.187*(144-60)] m steam = 3,719.7 kg steam Heat Evolved from Cell Growth (kcal) = 0.12 * Oxygen Uptake Rate (mmol/(h)) * Fermentation Time (h) =0.12 * mmol/h * h =2, kcal

40 Calculations - Separations Centrifuge #2 Mass Balance: ΣMass = Mass in – Mass out = 0 0 = 22, (kg/batch of lysed algae) – 6, (kg/batch of algae) – 16, (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) (kg/hr of air) – 63,295 (kg/batch waste water) -23,368 (kg/batch of conc. algae) (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) (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 Tray /6/ *0.092/1000*0.001 = 0.42 gal/hr Tray /6/ *0.092/1000*0.007 = 2.91 gal/hr Tray /6/ *0.092/1000*0.015 = 6.23 gal/hr Tray /6/ *0.092/1000*0.015 = 6.23 gal/hr HX-11 Oil into the Vacuum Vessel Q oil = mc p (∆T) Q oil = *0.55*( ) Q oil = BTU/hr Q Poil = mc p (∆T) T = 617- (109454/2900/.79) T = 569.2°F ∆T lm = [(T 1 – t 2 ) – (T 2 – t 1 )]/ln[(T 1 – t 2 )/ (T 2 – t 1 )] ∆T lm = [( ) – ( )]/ln[( )/( )] ∆T lm = 188.8°F Q = UA ∆T lm A = /60/188.8 A = 9.68 ft 2