Derrick R. J. Kolling March 16, 2010

Potential Algal Biofuel Applications in West Virginia

Derrick R. J. KollingDepartment of Chemistry

STaR SymposiumOctober 23, 2013

Derrick R. J. Kolling October 23, 2013

Talk Overview• Biofuels overview

– Why do we need biofuels?• The light reactions of photosynthesis

2H2O + 4h 4H+ + 4e- + O2

• Tools of the trade– O2 electrode– Fluorometer– EPR spectrometer– GC-MS

• Research projects – OEC photoassembly– Temperature dependence of molecular photosynthesis– Algal lipidomics– Biofuels from invasive algal species

Renewable EnergyBiofuelsSolar powerWind powerTidal powerHydropowerGeothermal

futurefarmers.com/survey/algae

epmb.berkeley.edu

Derrick R. J. Kolling October 23, 2013

Dismukes et al. (2008)

Derrick R. J. Kolling October 23, 2013

Photosynthesis

Derrick R. J. Kolling March 16, 2010

Carbon Fixation

Electrons(as NADH, NADPH,Reduced Ferredoxin)

Biomass

CO2

H2OO2

Protein Synthesis

e-

ATP

Source: Damian Carrieri

anaerobic

Fermentation

O2

H2

lipidprotein

carbohydrate

biodieselanimal feed

ethanol

CO2, organic acids

ATP

Lipid Synthesis

H2O

Derrick R. J. Kolling October 23, 2013

Dismantling photosynthesis from the organism to the molecules….

Light Reactions

Derrick R. J. Kolling October 23, 2013

Barber & Iwata (2004) Science

…from the molecular to the atomic level

Umena et al. (2011) Nature

Light Reactions

Derrick R. J. Kolling October 23, 2013

Research Tools• Oximetry • Fluorescence

• GC-MS • Electron Paramagnetic Resonance

0 5 10 15 20 25 30 35 40 45 500

100200300400500600700800900

BBY particles w/ out DCBQ

BBY particles w/DCBQ

Normalized BBYs w/out DCBQ

Temperature °C

O2

ev

olu

tio

n r

ate

(µ

mo

l O2

/mg

Ch

l a/h

)

Temperature dependence of oxygen evolution

PSII PSII w/DCBQ

PSII normalized

OEC Photoassembly

OEC Photoassembly

Temperature dependence of oxygen evolution vs. photoassembly

Photoassembled PSII at 28°C

Intact PSII

-Share a rate-limiting step

OEC Photoassembly

5 10 15 20 22 25 26 27 28 29 30 32 35 400

20

40

60

80

100

120Percent Recovery Oxygen Evolution

Temperature °C

Per

cen

t R

eco

very

-Protease activity explains dip

OEC Photoassembly

Findings

• Oxygen evolution and OEC photoassembly share a 28°C maximum

• OEC photoassembly and PSII oxygen evolution appear to share a rate-limiting step

• Thermal inactivation of PSII and apo-PSII occurs at temperatures greater than 28°C

• Thermal inactivation is partially due to protease activity (Deg and/or FtsH?)

OEC Photoassembly Application

www3.imperial.ac.uk

Artificial Leaf

www.ruhr-uni-bochum.de/h2design/profile/main.html

Bioinspired/biomimicked System

Algal Lipidomics

Comparison of lipid accumulation in photomixotrophically and heterotrophically grown Chlorella vulgaris cultures

Algal Lipidomics

[ [ [ [ [ [ [ [ [

0 1 2 3 54 6 7 8

Cultu

re1

2

3

4

5

6

Measured: -[Chl a] -turbidity -dry weight -lipid dry weight

Algal Lipidomics

0 1 2 3 4 5 6 7 8 90

5

10

15

20

25

30

Pho-tomixotrophicHeterotrophic

Time (Days

Tu

rbid

ity

(AU

)

0 1 2 3 4 5 6 7 8 90

5

10

15

20

25

30

Photomixotrophic

Heterotrophic

Time (Days)

Ch

l a

(mg

/ml)

Cell Density750nmChl a Expression

Derrick R. J. Kolling October 23, 2013

Algal Lipidomics

0 1 2 3 4 5 6 7 8 90

2

4

6

8

10

Series1

Time (Days)

Wei

gh

t (m

g)

Photomixotrophic Cells

Heterotrophic Cells

Photomixotrophic Lipids

Heterotrophic Lipids

Dry Weight Growth Curves and Corresponding Dry Lipid Weights

C 18:2(n-6): linoleic acid

C 18: stearic acid

C 18:1(n-9): oleic acid

C 16: palmitic acid

Algal Lipidomics

• Photomixotrophically grown cells produce 2X as many lipids as do heterotrophically grown cells

• Photomixotrophically grown cells reach stationary phase and higher biomass sooner

• Cells produce palmitic, stearic, oleic, and linoleic acids

Findings

Algal Lipidomics Application

Chisti (2007) Biotech. Advances

Derrick R. J. Kolling October 23, 2013

• Temperature Dependence of PSII-James Board, Hope Cook, Matt Thompson, Shane Kagen, Jordan Hilgeford, Justin Erwin• OEC photoassembly-James Board, Hope Cook, Ben Blodgett, Matt Thompson, Shane Kagen,

Jordan Hilgeford, Chase Turner, Ben Weiner• Algal lipidomics-Ben Woodworth, Tony Stephenson, Rebecca Mead, Courtney Nichols,

Prof. Leslie Frost (MU)• Bioethanol from invasive algal species-Kevin Militello, Shaheed Elhamdani

Acknowledgements

Acknowledgements

Funding

This material is based upon work supported by the National Science Foundation under Cooperative Agreement Award number EPS-1003907 and CHE-1229498.