Biofuel Production from Algae Species/Strain Selection

The Power of Algae: Photobioreactor Design and Production Strategies for Biofuel Production Joel L. Cuello, Ph.D.

Department of Agricultural and Biosystems Engineering The University of Arizona DOE Project Hybrid Solar and Electric Lighting System Hybrid Solar and Electric Lighting System Hybrid Solar and Electric Lighting System Hybrid Solar and Electric Lighting System Hybrid Solar and Electric Lighting System U.S. Department of Energy (DOE)

CO2 Assimilation by Algae

David Bayless and Joel L. Cuello Ohio University and The University of Arizona Hybrid Solar and Electric Lighting System

Solar collector

Optical fiber cable

Light emitting panel Bioreactor

Flue gas injection Biomass Hybrid Solar and Electric Lighting System Hybrid Solar and Electric Lighting System Hybrid Solar and Electric Lighting System Algae Growth Rates as Function of CO2 Levels

0.7

0.6 250 Elevated CO 200 2 0.5 100 (5% CO2 supplemented) 250 CO2 200 CO2 0.4 100 CO2

0.3

Dry Weight (g/L) 0.2 Ambient CO2

0.1

0 -2 0 2 4 6 8 10

Number of days CO2 Assimilation by Algae

15 Carbon Assimilation (mgC/L day) 10

5 5% H

5% L 0 250 Ambient 200 100 Light Conditions (mmol sec -1 m-2) Algae Algae: Major Advantages (1) renewable energy source

(2) potential for reduction of emissions from power plants

(3) much higher productivity than traditional fast-growing energy crops

(4) less area required than traditional crops when grown in photobioreactors Algae: Major Advantages

(5) production in photobioreactors prevents potential degradation of soil and groundwater

(6) non-potable water can be used, aiding in wastewater treatment and utilizing non-productive areas

(7) production of economically valuable chemicals Algae: Major Advantages

(8) Energy feedstock that does not compete with food or feed! Products from Microalgae

Chemical Usage Approx. Value ($/kg) Astaxanthin Salmon aquaculture, pigment >2500 b-carotene Food supplement >500 Phycobiliproteins Medical diagnostics >10,000 Food colors >100 Health supplements Dietary supplement 20-10 Xanthophyll Chicken feeds 200-500 Fish feeds 1000 Algae: Biodiesel Yield (L/ha-yr)

Soybeans 446 Rapeseed 119 Mustard 1300 Jatropha 1892 Palm Oil 5950 Algae (Low) 45000 Algae (High) 137000 Ours (High) 132,300! Which Algae do We Work with?

•Those accumulating hydrocarbons •Those accumulating fatty acids •Those accumulating starch •Those producing hydrogen gas

Biofuel Production from Algae Species/Strain Selection

Mass Production of Algae

Harvesting

Dewatering

Product Extraction/Processing Two Ways to Mass Produce Algae

Open Ponds

Photobioreactors Open Pond System Open Pond System

http://www.veggievan.org/downloads/articles/Biodiesel%20from%20Algae.pdf Photobioreactor Design Photobioreactor Controlled Light Nutrients

CO2 Mixing Algae Culture Density pH Temperature Flow Rate etc. Photobioreactor Designs Photobioreactor Design

Biofuel Production from Algae Species/Strain Selection

Mass Production of Algae

Harvesting

Dewatering

Product Extraction/Processing

Hydrodynamic-Based Design of Algae Photobioreactors for Biofuel Production

Michael Mason, In-Bok Lee and Joel L. Cuello The University of Arizona B. braunii growth optimization

50.000 45.000

40.000 mix., CO2, 200umol 35.000 mix, no CO2, 200umol 30.000 mix., CO2, 150umol mix., no CO2, 150umol 25.000 no mix., CO2, 200umol

F.W. (g/L) 20.000 no mix., no CO2, 200umol 15.000 no mix., CO2, 150umol 10.000 no mix., no CO2, 150umol 5.000 0.000 0 2 4 6 8 10 Time (days) Criteria for Algae Photobioreactors Delivery of Light

Delivery of CO2

Delivery of Nutrients

Optimal Culture density

Mixing/Hydrodynamic Conditions!!! Design and Scale Up Strategy

Axial Dispersion Coefficient

Vessel Dispersion Number Photobioreactor Bodenstein Number hydrodynamic conditions Reynold’s Number

Mixing Time

Identify and select set of hydrodynamic Photobioreactor conditions that translate into optimal algae growth rate/ productivity algae growth rate/productivity Use of RTD to Determine Hydrodynamic Parameters Determined Residence Time Distributions in 2 sizes of External Air-Lift Photobioreactor at 3 volumetric flow rates at coarse and fine bubble sizes.

Volume (4.4. L) of the small reactor whose diameter is 2 in is roughly half the volume (10.2 L) of the big reactor whose diameter is 3 in. Example RTD Results for External Air-Lift

At Volumetric Flow Rate = 4.7 L/min with Coarse Bubbles (0.25” dia)

[NaCl] [NaCl] (g/L) 1.8 1.400 1.6 1.200 1.4 1.000 1.2 1 0.800 0.8 [NaCl] 0.600 [NaCl] (g/L) 0.6 0.400 0.4 0.2 0.200 0 0.000 0 200 400 600 0 50 100 150 200

At 2 in diameter At 3 in diameter RT = 55 s RT = 97 s Dispersion No. = 0.024 Dispersion No. = 0.058 Bodenstein No. = 41.7 Bodenstein No. = 17.4 Reynold’s No. = 1961 Reynold’s No. = 1311 Mixing Time: Air-Lift

NaCl Electrical Injection Conductivity Port Sensor

Air Flow Meter Mixing Time: Bubble Column

Air Flow Meter

NaCl Injection Port Mixing Time: Bubble Column

Electrical Conductivity Sensor Mixing Time Results

LC7/25_1 LC7/50_1

1.2 1.2

1 1

0.8 0.8

0.6 0.6 EC(mS/cm) 0.4 0.4

0.2 0.2 0 0 500 1000 1500 2000 2500 3000 0 seconds 0 500 1000 1500 2000 2500 3000

Control, 7 gpm, 25 vs. 50 cfh

Bubble Column reactor

15 Algae growth as bbl col function of 10 LOW F.W. (g/L) hydrodynamic 5 bbl col MED conditions 0 0 5 10 15 20 Day

Air-Lift reactor

15 air lift 10 LOW F.W. (g/L) 5 air lift MED 0 0 5 10 15 20 Day B. braunii growth optimization

50.000 45.000

40.000 mix., CO2, 200umol 35.000 mix, no CO2, 200umol 30.000 mix., CO2, 150umol mix., no CO2, 150umol 25.000 no mix., CO2, 200umol

F.W. (g/L) 20.000 no mix., no CO2, 200umol 15.000 no mix., CO2, 150umol 10.000 no mix., no CO2, 150umol 5.000 0.000 0 2 4 6 8 10 Time (days) Flask growth experiment

25.0 mixing, no CO2, 200umol 20.0

mixing, no CO2, 15.0 150umol

10.0

F.W.(g/L) no mixing, no 5.0 CO2, 200umol

0.0 0 1 2 3 4 5 6 7 8 9no mixing, no CO2, 150umol -5.0 day

Successful scale-up, higher productivity in photobioreactors A Third Way to Mass Produce Algae Algae Aquaculture n Algae represent the largest aquaculture crop on a global basis n Algae are a major component of diet in Asia Goal

n Design microalgae production systems based on aquacultural growing systems n Lower cost n Small-scale and distributed production systems n Realistic promotion of economic diversification in U.S. rural areas Coupled Aquaculture and Hydroponics Coupled Aquaculture Effluent and Algae Aquaculture and Algae Biofuel Photobioreactor Design Integrated Algae/Fish-Feed Aquaculture

Biofuel

Fish Feed Integrated Algae-Fish Aquaculture Fish Feed Algae Residues Algae

Treated Fish Waste/Water Biofuel Water

Fish Body Residues Fish Fish Meat Aquaculture and Algae Biofuels n Aquaculture producers are well- positioned to produce fish meat, fish/animal feed, and algae oil. Production Strategies Production Strategy:

Composite Lighting for Algae Biofuel Production HYBRID SOLAR AND ELECTRIC LIGHTING (HYSEL) FOR SPACE LIFE SUPPORT

J.L. Cuello, T. Nakamura, D. Larson, K. Jordan, E. Ono and H. Watanabe

Department of Agricultural and Biosystems Engineering The University of Arizona National Aeronautics and Space Administration (NASA)

Grant No. NAG10-0255

Concentrator’s Spectral Output

2500

2000

1500

1000

500 Relative Intensity .

0 300 400 500 600 700 800 900 Wavelength (nm) Fiber 1 Fiber 2 Fiber 3 HYSEL System -- LED HYSEL System -- LED HYSEL System’s Spectral Output

HYSEL - LED

140 200

120 HYSAL1 HYSAL2 100 150 HYSAL3

80 100 60 Relative Intensity 40 Relative Intensity 50 20

0 300 400 500 600 700 800 900 0 Wavelength (nm) 300 400 500 600 700 800 900

Wavelength (nm) HYSEL System -- XMH HYSEL System -- XMH HYSEL System’s Spectral Output

HYSEL - XMH

40 350

35 300 30 250 25 200 20 150 15 Relative Intensity

Relative Intensity 10 100

5 50

0 0 300 400 500 600 700 800 900 300 400 500 600 700 800 900 Wavelength (nm) Wavelength (nm) Lighting Profiles

HYSEL 322 322 PPF 3.9 hr 3.9 hr 30 30

0 24 24 48

HPS 194 194 PPF

9.5 hr 9.5 hr 0 24 24 48 Results - Dry Weight

2 n=15

1.5

0.5 DRY WEIGHT PER PLANT (g)

0 CompositeHYSAL ConventionalHPS LIGHTING SYSTEM Results – Chlorophyll Content

n=15

0.75

n=15

0.5

0.25 CHLOROPHYLL (mg/g FW)

0 CompositeHYSAL ConventionalHPS LIGHTING SYSTEM Composite Lighting Applied to Algae

Chlamydomonas Botryococcus Lettuce reinhardtii braunii

Takanori Hoshino and Joel L. Cuello The University of Arizona Composite Lighting Applied to Algae

Conventional 300 Dark Period 12 hours per day 200 µmol m-2 s-1 200 PPF average 200 µmol m-2 s-1 12 hours (During Light Period)

100

Dark 0 0 12 24

Composite 300 Dark Period 0 hours per day

-2 -1 200 175 µmol m s PPF average 100 µmol m-2 s-1 (During Light Period) 12 hours 100 25 µmol m-2 s-1 0 0 12 24 Chlamydomonas reinhardtii Composite Lighting Applied to Algae

Nutrition Light Cycle

Control Photoautotrophic 12 hours: 200 µ mol m-2 s-1 With 5% CO2 provided 12 hours: Dark Composite Lighting Photoautotrophic 12 hours: 175µ mol m-2 s-1 -2 -1 With 5% CO2 provided 12 hours: 25 µ mol m s Results - Chlamydomonas

2.0 Nutrient Limitation Cont Treat 26%

1.5 suspension) 1 -

1.0

0.5 50 – 250 hours y = 0.0061x – 0.0223

Dry Cell Weight (g DCW L y = 0.0047x – 0.0085

0.0 Bars: Standard Deviation with N = 5 0 100 200 300 400

Time (hour) Results - Chlamydomonas

Cont 45 Nutrient Limitation Treat 40

30 63%

Starch Content (%: g glucose/g CDW) 5 Bars: Standard Deviation with N = 5 0 0 50 100 150 200 250 300 350 400 Time (hour) Conclusion

For a fixed quantity of light energy, Composite Lighting resulted in significantly greater algae growth compared with that for Conventional Lighting. Conclusion

Equal moles of photons do not necessarily result in equal growth in green algae Growth of Algae in Wastewater

Sara Kuwahara and Joel L. Cuello The University of Arizona Growth of Algae in Wastewater Algae Wastewater Treatments

Nutrients Reclaimed Reclaimed Diluted 50% Reclaimed Modified Chu-13 (mg/L) water water + 50% P modified Chu-13 water + 100% and 50% N P and 100% N

Treatment A B C D E Calcium 48.70 48.70 14.976 8.70 29.952 Carbon 218.00 218.00 21.696 218.00 43.392 Chloride 90.00 90.00 13.263 90.00 26.525 Magnesium 7.80 7.80 9.859 7.80 19.718 Nitrogen 20.60 25.632 25.632 51.263 51.263 Phosphate 12.00 12.0 7.112 14.224 14.224 Sodium 116.00 142.00 0.000 116.00 00.000 Sulfate 106.00 106.00 25.229 106.00 50.459 Growth of Algae in Wastewater Growth of Algae in Wastewater

Dry Weight (dry g/ g dry starting weight) control 4.0

3.5

3.0

2.5 100% sw

2.0

1.5

1.0 g dry weight/ starting weight 0.5

0.0 0 20 40 60 80 100 120 140 160 Time (hour)

Average A Average B Average C Average D Average E Growth of Algae in Wastewater

Total Chlorophyll (g/L) 100% sw 25.0

20.0

15.0 control

10.0

5.0

0.0 0 20 40 60 80 100 120 140 160

Average A Average B Average C Average D Average E Growth of Algae in Wastewater

Total Nitrate (mg/L) 70.0

60.0

50.0

40.0

30.0

Total nitrate (mg/L) 20.0

10.0

0.0 0 20 40 60 80 100 120 140 160 Time (hours)

Average A Average B Average C Average D Average E Growth of Algae in Wastewater

Total Organic Phosphate (mg/L) 80.0

70.0

60.0

50.0

40.0

30.0

20.0 Total Organic Phosphate (mg/L) 10.0

0.0 0 20 40 60 80 100 120 140 160 Time (hours)

Average A Average B Average C Average D Average E Hydrogen Gas Production from Algae

Takanori Hoshino and Joel L. Cuello The University of Arizona Acknowledgments Acknowledgments