Optimizing Lipid Production by Planktonic Algae: LIPIDO Kristian Spilling Finnish Environment Institute, SYKE Project leader: Timo Tamminen

Micro-algae ● Aquatic, free-floating, unicellular or filamentous (phytoplankton) ● Some are very fast-growing ● Cells contain minimal amounts of structural component, but mainly of proteins, lipids (oil compounds) and carbohydrates

10 µm

Why planktonic algae? – biology and biochemistry ● Harvest cycle ○ Forest biomass: years to decades ○ Field biomass: months ○ Microalgae: days, even hours ● Biomass composition ○ Plankton: oil compounds (lipids) can be very high (even 40-60%) – no structural compounds ● Production per area ○ Higher to very much higher (2-10 times), compared with the best terrestrial crops

Potential benefits of algae ● Algae can be grown in a way that do not compete for fertile land ● Has potentially very positive energy / carbon balance ● Salt or wastewater can be used as the base for culturing ● Can be coupled with CO2 producing industry ● High potential for growth ● Potentially high lipid yield

Algal lipids as feedstock for biodiesel

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Partners - Finland ● Finnish Environment Institute (SYKE) ○ Timo Tamminen ○ Kristian Spilling ○ Jukka Seppälä

● Algal growth experiments, environmental parameters ● VTT, Technical Research Centre of Finland ○ Kirsi-Marja Oksman-Caldentey ○ Heiko Rischer ○ Dagmar Enss

● Lipidomics, overall biorefinery concept, downstream and sidestream processing

Partners - Norway

● Norwegian University of Science and Technology (NTNU), ○ Olav Vadstein ○ Yngvar Olsen ○ Matilde Skogen Chauton (Post Doc)

● Algal stoichiometry, growth yield and reactor technology ● University of Oslo ○ Tom Andersen ○ Per Færøvig (Post Doc)

● Algal growth response and optimization

Partners - Germany

● Ludwig Maximilian University (LMU) ○ Herwig Stibor ○ Maria Stockenreiter (PhD student) ○ Florian Haupt (PhD student)

● Algal stoichiometry and fatty acid composition

Partners - Iceland

● Blue Lagoon ○ Ása Brynjólfsdóttir ○ Halldór G. Svavarsson (Reykjavik Univ.) ● Geothermal energy / CO2, algal cultivation

Project focus Optimizing Lipid Production by Planktonic Algae

Downstream processing:

Growth

Lipids

Harvest

Lipid extraction Water recycling Biofuel production Remaining biomass

Project focus, division of labor

Iceland Thermophilic algae

Norway Marine algae

Germany Freshwater algae

CO2

Finland Brackish water algae

Project goals ● To screen for the most promising algal species for temperate environments ● To optimize their growth and lipid yield as functions of growth conditions ● To test the practical applicability of coupling algal culturing to CO2 emission mitigation ● To screen commercially interesting by-products from biomass of selected species

12

Algal physiology

Vegitative growth Algae Algae

Inorganic nutrients e.g. N & P

Algae

Carbohydrates Storage Lipids

Activities – Equipment development Oslo Trondheim

0

C18:4?FFA

C18:3n3FFA

C18:3n6FFA

C18:2FFA

20

C18:1n7FFA

Diatoms

C18:1n9FFA

Isochrysis

C18:0FFA

C22:6FAME (16:3FFA?)

C22:5FAME

C16:2FFA

Chlamydomonas

C16:1FFA

Pavlova

C16:0FFA

C20:5FAME

C20:3n3FAME

C20:4n6FAME

C20:3n6FAME

C14:0FFA

C18:4?FAME

30

C18:3n3FAME

Thalassiosira pseudonana

C18:3n6FAME

Thalassiosira baltica

C18:2FAME

C18:1n7FAME

C18:1n9FAME

C18:0FAME

C16:3FAME

C16:2FAME

50

C16:1FAME

C16:0FAME

40

C15:0FAME

60

C14:0FAME

FA. % of total FA

Activities: screening

70

Monoraphidium Chlorella

Dinophyta

Melosira Chaetoceros Haptophyta Gymnodinium

10

Activities: growth optimization

Growth rates [d-1] Chaetoceros wighamii

Thalassiosira baltica

450 400

Irradiance

350 300

0,0 0,1 0,2 0,3 0,4 0,5 0,6 0,7 0,8

250 200 150 100 50 5

10

15

temperature sp. Pseudochattonella

20

5

10

15

temperature

20

Activities: lipid accumulation

Chaetoceros 450 400

Irradiance

350 0,8 1,0 1,2 1,4 1,6 1,8 2,0 2,2 2,4 2,6

300 250 200 150 100 50 3

4

5

6

7

8

temperature

Total FA as % of DW

9

10

11

Activities: lipid accumulation

Chaetoceros wighamii

Thalassiosira baltica

Alternative CO2 sources / CO2 mitigation

19

Alternative CO2 sources / CO2 mitigation

20

Effect of different CO2 concentrations

CO2 depleted CO2 repleted 21

By-products, antimicrobial effect of algae

22

Outcome: articles (Sept 2011) ●

●

Rischer, H (2009) Photosynthetic microorganisms as a future source of energy. In: K. Larjava (ed.), Energy Visions 2050, WS Bookwell Oy, Porvoo, 246-247. Packer A, Li Y, Andersen T, Hu Q, Kuang Y, Sommerfeld M (2011) Growth and neutral lipid synthesis in green microalgae: a mathematical model Biores.Technol. 102: 111-117. Spilling K, Seppälä J, Tamminen T (2011) Inducing auto-flocculation in the diatom Phaeodactylum tricornutum through CO2 adjustment. J Appl Phycol. DOI 10.1007/s10811-010-9616-5 Stockenreiter M., Graber A.-K., Haupt F. and Stibor H. (2011). The effect of species diversity on lipid production by micro-algal communities. J Appl Phycol. DOI: 10.1007/s10811-010-9644-1. Enss D, Seppälä J, Spilling K, Tamminen T, Oksman-Caldentey K-M, Rischer H. Growth phase changes in lipid profiles of twenty phytoplankton species adapted to different temperatures. Submitted research article. Spilling K., Seppälä J. Photobiology and lipid metabolism of algae. Submitted book chapter.

●

Additioal 4-5 papers related to the work in LIPIDO in preparation

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Outcome (Sept 2011) ● Researcher training ○ 3 PhD students ○ 2 MSc thesis ○ 3 student projects ● Researcher mobility ○ 2 common experiments ● New research projects building on the results of LIPIDO

Conclusion ● Key focus: growth and lipid optimization ○ New tools and methods being developed ○ Screening for lipids ○ Growth and lipid optimization ○ What stress factors increase the lipid yield? ● Other topics ○ Harvesting ○ Potential for using CO2 from power plant ○ Interesting bi-products

Thank you for your attention

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