Derrick R. J. Kolling March 16, 2010

Enhancing Algal Biodiesel Through the Use of Waste Products

Derrick R. J. Kolling

Department of Chemistry

Renewable Energy in West Virginia

June 5, 2014

Derrick R. J. Kolling June 5, 2014

• Biofuels research justification – Why do we need biofuels?

• Algal biodiesel – TAGs

• Research projects – Photomixotrophically grown algae – Using glycerol to increase algal biodiesel yields

Overview

Renewable Energy

Biofuels

Solar power

Wind power

Tidal power

Hydropower

Geothermal

futurefarmers.com/survey/algae

epmb.berkeley.edu

Derrick R. J. Kolling June 5, 2014

Dismukes et al. (2008)

Biofuel Footprint

Derrick R. J. Kolling June 5, 2014

Available Biomass

Photosynthesis

Derrick R. J. Kolling March 16, 2010

Carbon Fixation

Electrons (as NADH, NADPH, Reduced Ferredoxin)

Biomass

CO2

H2O O2

Protein Synthesis

e-

ATP

Source: Damian Carrieri

anaerobic

Fermentation

O2

H2

lipid protein carbohydrate

biodiesel animal feed

ethanol

CO2, organic acids

ATP

Lipid Synthesis

H2O

Algal Biofuel

Chisti (2007) Biotech. Advances

Algal Biofuel

Scott et al. (2010) Current Opinion in Biotechnology

Algal Biofuel

Comparison of lipid accumulation in photomixotrophically and heterotrophically grown Chlorella vulgaris

[ [ [ [ [ [ [ [ [

0 1 2 3 5 4 6 7 8

Cultu

re

1

2

3

4

5

6

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

0

5

10

15

20

25

30

0 1 2 3 4 5 6 7 8 9

Turb

idity

(AU

)

Time (Days

PhotomixotrophicHeterotrophic

0

5

10

15

20

25

30

35

0 1 2 3 4 5 6 7 8 9C

hl a

(mg/

ml)

Time (Days)

PhotomixotrophicHeterotrophic

Cell Density750nm Chl a Expression

Derrick R. J. Kolling June 5, 2014

0

2

4

6

8

10

0 1 2 3 4 5 6 7 8 9

Wei

ght (

mg)

Time (Days)

Series3Series4Series1Series2

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

Derrick R. J. Kolling June 5, 2014

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

• This increased to 5X under N deprivation (50 %)

• Photomixotrophically grown cells reach stationary phase and higher biomass sooner

• Cyclic electron transfer

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

Derrick R. J. Kolling June 5, 2014

Findings

Using glycerol to maximize lipid production in Chlorella vulgaris

Derrick R. J. Kolling June 5, 2014

Derrick R. J. Kolling June 5, 2014

Working hypothesis: Increased glycerol availability will increase the production of TAGs in C. vulgaris

Scott et al. (2010) Current Opinion in Biotechnology

Radakovits, et al. Eukaryotic Cell

Extracellular matrix Glycerol

Glycerol

Glycerol transporter

Glycerol kinase

Modified from Radakovits, et al. Eukaryotic Cell

Derrick R. J. Kolling June 5, 2014

Stationary phase cells were exchanged into control or N-deplete medium that contained 1% glycerol

• N-deprived cells accumulated ~%50 biomass of control

• %20 of biomass in N-deprived cells was lipids vs. %15 in the control

Derrick R. J. Kolling June 5, 2014

Findings

• Algal biodiesel -Ben Woodworth, Tony Stephenson, Rebecca Mead, Courtney Nichols, Morgan Stickler, Kasey Stickler, Mallory McDonald, Aaron Holland

•OEC photoassembly -James Board, Hope Cook, Ben Blodgett, Matt Thompson, Shane Kagen, Chase Turner, Ben Weiner, Justin Erwin, Jordan Hilgeford

• Bioethanol from invasive algal species -Kevin Militello, Shaheed Elhamdani

Acknowledgements

Acknowledgements

Funding

Derrick R. J. Kolling June 5, 2014

Research Tools • Oximetry

• Fluorescence

• GC-MS

• Electron Paramagnetic Resonance

OEC Photoassembly Application

www3.imperial.ac.uk

Artificial Leaf

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

Bioinspired/biomimicked System

Wijffels, R.H. and Barbosa, M.J.2010 Science