Microalgae-based biofuel technology

Peer Schenk
School of Biological Sciences Faculty of Science The University of Queensland Australia p.schenk@uq.edu.au 6 October 2009 2009 Microalgae Technology Conference Taiwan

IMPORTANCE OF CO2 SEQUESTRATION

RENEWABLE ENERGY SOURCES

SOLAR: 126,000 TW

Plants

GEOTHERMAL: 92,000 TW-YR GEOTHERMAL: 92,000 TW

Renewable, CO2-free energy sources

TIDAL POWER: 0.1 TW TW-YR WAVE: 2 TW-YR TW WIND: 5 TW-YR TW OCEAN THERMAL: 10 TW-YR TW BIOMASS: 172 TW-YR TW

vs. current energy demand

WORLD ENERGY DEMAND: 13 TW

Search for renewable fuels

increased attention on climate change decline in fossil fuel reserves fuels account for of global energy demand increasing cost environmental damage need for sustainable biofuel production without competing for arable land

BIO-FUEL PRODUCTION: PHOTOSYNTHESIS IS CENTRAL

1. DRIVES FIRST STEP IN THE CONVERSION OF LIGHT TO CHEMICAL ENERGY 2. PRODUCES FEEDSTOCKS FOR FUEL SYNTHESIS

3. BUT CONVENTIONAL CROPS HAVE ~1-3% PHOTOSYNTHETIC EFFICIENCY 4. INCREASING PHOTOSYNTHETIC EFFICIENCY INCREASES ECONOMIC VIABILITY

Why Algae biofuels?

Increasing demand for Biofuels
Fossil fuel depletion / Effect on Climate change Transport fuels: 66 % of total energy use biodiesel: high energy content and easy to process.

Problems with Terrestial Biofuels:

Large land requirements (low energy conversion efficiency) Only possible on scarce arable land
Competition with food production Deforestation

High water consumption (runoff, evaporation) Inefficient nutrient consumption (runoff N2O emissions)

Advantages of Algae Fuel

No Arable land required High yields due to:
High biomass density no roots, leaves, stems High growth rates (DT as fast as ~3.5 h) High tryacylglyceride (TAG) content (~20 - 70 %)

Low water consumption:

Seawater/ agricultural runoff water / waste water. No loss due to runoff No evaporation in closed bioreactor

Efficient use of nutrients CO2 capture possible Residue methane/ethanol, cattle feed,

The ultimate energy cycle

CO2 & NOX

Hydrogen Production Waste from Sewage plant Biodiesel & Bioethanol Production

Waste to Power plant

Algae Biotechnology

Biohydrogen

Agrichar (carbon sequestration)

http://www.cmtevents.com

The University of Queensland Algae Biodiesel Project

Collection & culture of microalgae freshwater, brackish, marine environments (SE Queensland) Pure culture Identification (microscopy/ribosomal DNA sequencing) Selection criteria Growth rate Biomass Lipids Flocculation/oil extraction Algae breeding Midscale outdoor cultivation

Microalgae collection - BR

Pure culture

identified microalgae
Culture ID BR2 BR8a BR8b AR1, BR17, PAG BR19 BR25 BR30 SD3 ESK TOWER P1
Timmins, et al., 2008 Eukaryotic Cell

Given Name Scenedesmus sp. isolate BR2 Scenedesmus sp. isolate BR8a Desmodesmus sp. isolate BR8b Chlorella ducis Desmodesmus tenuis Desmodesmus irritus Chlorella caelum Chlorella optimates Chlorella sp. Esk Tower Desmodesmus viridis

Accession Number EU729732 EU729729 EU729730 EU502834 EU502832 EU502835 EU502833 EU502837 EU729731 EU502836

Nile red staining

Nile red stained culture x400 uv fluorescence

Lipid analysis (TLC and GC-MS)

Quality of fuel
Fatty Acid C10:0 C12:0 C14:0 C14:1 C16:0 C16:1 C16:1 iso C16:2 C16:3 C18:0 C18:1 C18:1 iso C18:2 C18:3 C18:4 C18:5 C20:3 C20:4 TOTAL 1 99.99 100.00 99.99 100.00 99.99 99.99 11.07 31.28 7.23 3.55 3.53 22.09 3.23 12.53 3.22 15.13 13.13 5.82 8.41 1.45 1.55 2.16 7.59 18.45 5.83 12.68 4.47 4.55 61.94 5.24 15.12 60.87 4.04 69.89 4.07 46.41 6.92 BR2 3.02 1.44 4.75 11.84 BR8 BR20 6.55 BR21 3.14 BR30 3.19 ORI1 ORI4 ESK TOWER 3.81 (polyunsaturated

2.04Low 1.27 PUFA

fatty acids)
3.52

Less oxidation 2.19 2.31 26.83 Long term storage 35.62 21.86

5.65

5.51High PUFA
1.51 3.63 0.83 1.79

8.54

10.8

10.25

5.33 4.27 Good cold flow 2.68 properties 7.42 3.61 3.76 6.59 Overcomes cold filter 1.41 13.26 plugging point (CFPP) 7.35 6.97

7.79An Ideal mix

19.91 27.4(Schenk,25.85 2008 Bioenergy Research) et al.,

16:1, 18:1 and 14:0 in the ratio 5:4:1

99.99 99.99

Search for inexpensive nutrient sources

Soil extract medium

Wastewater UQ Tennis courts AWMC

Algae testing and media optimisation

Raceway pond
(low nutrient)

10-Litre Bioreactor

Split system design for synchronised TAG induction and avoidance of contamination
Bioreactor
(high nutrient)

sun
Batch transfer
Clean water

Raceway pond
(low nutrient)

Wastewater CO2

Split system design for synchronised TAG induction and avoidance of contamination
Laboratory inoculum
(exponential growth)

Bioreactor
(continuous exponential growth)

Raceway pond
(low nutrient)

Nutrients, Wastewater, Seawater, CO2

Open raceway ponds

TAG Induction, Extraction and Biodiesel Synthesis

Open raceway ponds
(low nutrient water)

Oil extraction
(foam fractionation)

Biodiesel synthesis (transesterification)

Harvest after 3 days and replace with culture

ALGAE BREEDING Adaptive evolution of microalgae to select for high TAG cells
yellow

red

BR2 evolved for high TAG production (synchronised induction)

Raceway pond
(low nutrient)

Nutrients, Wastewater, Seawater, CO2

Raceway pond
(low nutrient)

Bioreactor team

UQ Algae biodiesel team

Coordination
Peer Schenk Skye Thomas-Hall Eugene Zhang Stephanie Ewert Bart Nijland Priyanka Nayak Matthew Timmins Philipp Keymer Fauzi Haroon Alex Metcalf

Algae Breeding/Adaptive Evolution

Adam Posthuma Yamini Kashimshetty Miklos Deme Kalpesh Sharma Sourabh Garg Liguang Wang Steve Welsh Christopher Beavon Stephen Su

Algae collection

Midscale Outdoor Cultivation

PUFA analysis Media

Oil Extraction/Biodiesel Synthesis

Grant applications

North Queensland & Pacific Pty Ltd

Photo-Biological Hydrogen Production

in the Green Alga Chlamydomonas reinhardtii

MOLECULAR BIOLOGY GENETIC SCREENING

Focus: Developing economic solar-powered H2 production from H2O using engineered green algal cells.

Solar Solar Energy Energy Algae Algae

22H2O H2 O O2 O2

PHOTOSYNTHESIS NATURAL SURVIVAL MECHANISM

22H2 H2

BIOCHEMISTRY

MOLECULAR BIOLOGY GENETIC SCREENING

Photosynthetic H2 production in green algae

Small scale Experimental set-up:
Measurement of gas purity via GC

Gas-volume determination

Monitoring Dissolved Oxygen and pH

Sampling and injecting possible

H2 produced can power a fuel cell car

Photosynthetic H2 production in green algae

MOLECULAR BIOLOGY GENETIC SCREENING

Mutant with high efficiency H2 production

Stm6 Stm6

Starch Store Starch Store

WT WT

1. H20 > H+ + e- > H2 2. STARCH > H+ + e- > H2

BIOCHEMISTRY: SOLAR POWERED H2 PRODUCTION FROM H2O

-S

Hydrogen production during anaerobiosis

Microarray Analysis

4650 genes differentially expressed 644 genes significantly induced or repressed.

Rearrangement of photosynthetic antenna

Skye Thomas-Hall, Anh Vu Nguyen, Alize Malno, Matthew Timmins, Evan Stephens, Jan H. Mussgnug p.schenk@uq.edu.au