Download - Catalyst - University of Washington

SUPERIOR ALGAL OIL PRODUCTION THROUGH
ACCELERATED EVOLUTION
A proposal prepared by
Team Algae Rules
Gregory Rowe
James Song
Eric Stephanson
James Zajac
for
Mike Daly
BP America Inc.
501 Westlake Park Blvd
Room 25. 157
Houston, TX 77079
July 22, 2011
ALGAE RULES
University of Washington
Seattle, WA 98195
7/22/2011
Mike Daly
Executive Vice President, Exploration
British Petroleum
501 Westlake Park Blvd
Room 25, 157
Houston, TX, 77071
Mike Daly,
The oil content of algae is useful as a source of the renewable energy, known as biofuels. However,
much of the production cost is related to removal of the oil from the algae resulting from expensive
extraction equipment and rising technology. In our submitted proposal, Superior Algal Oil
Production through Accelerated Evolution, Algae Rules believes that by genetically altering an
appropriate algae strain to produce twice the amount of oil content as well as retaining its ability to
be processed with the same energy input, production revenue could be significantly increased.
In 8 months, we propose to engineer a strain of algae to double its oil content for an estimated cost
of $473,000. We will select a candidate alga and identify target DNA sequences involved in oil
production. Then using current genome engineering technology, we will modify and evolve it to
maximize oil content. After it is inspected for performance, the alga will be made available to the
BP Company as well as existing algae biofuel producers.
We are eager to answer any questions or concerns that you may have about our proposal. We will
contact you next week to inquire on the current status of the proposal. Feel free to contact us
anytime at [email protected] or 1-800-123-4567. We hope to collaborate with BP with this
promising project to secure a sustainable future of biofuels.
Best Regards,
Algae Rules
Gregory Rowe, Eric Stephanson, James Song, James Zajac
CONTENTS
ILLUSTRATIONS .......................................................................................................................................................................1
Tables............................................................................................................................................................................. 1
Figures ........................................................................................................................................................................... 1
EXECUTIVE SUMMARY ........................................................................................................................................................2
SUPERIOR ALGAL OIL PRODUCTION THROUGH ACCELERATED EVOLUTION............................... 3
Introduction ................................................................................................................................................................ 3
Problem Statement .................................................................................................................................................. 3
Background Of The Problem ................................................................................................................................ 7
Personnel Information .......................................................................................................................................... 5
Purpose ......................................................................................................................................................................... 5
Scope .............................................................................................................................................................................. 5
Format ........................................................................................................................................................................... 6
Technical Information ............................................................................................................................................. 6
Project Work Summary .......................................................................................................................................... 7
Cost Information ....................................................................................................................................................... 8
Summary And Conclusion ..................................................................................................................................... 8
APPENDICES ........................................................................................................................................ 9
References ................................................................................................................................................................... 9
ILLUSTRATIONS
TABLES
1. Proposed Itemized Budget………….........................................................................................................p. 8
FIGURES
2. Figure 1: Origin Oil’s Single Step Oil Extraction Method…………………………………………..p.4
3. Figure 2: Project Timeline………………………………………………………..……………………………..p.7
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EXECUTIVE SUMMARY
As of now, attempts of replacing fossil fuels with renewable energies, specifically algae biofuels, are
struggling to expand to large scale operation. Many oil extraction methods have been experimented
via pilot projects, searching for a feasible method to upscale. However, the problems that occur
with modern extraction methods are that they are energy intensive and expensive. Moreover,
many companies fall short of obtaining potential profits from poor selection of algae strains.
Algae are processed in the three simple steps: rupturing the alga’s cell wall with a mechanical press
or expeller, removing the oil from the algae biomass with a solvent, and separating the oil from the
solvent by distillation. Most of the cost comes from extracting the oil from the algae, as much as
$1.80 per kg. With the price of biodiesel currently is $32/ gallon, current efforts to secure a
sustainable production of algae seem distant.
Algae Rules believes there is a feasible solution to the problem at hand: genetically alter an
appropriate algae strain that will produce twice the amount of oil for the same amount of energy
input. This will provide algae biofuel producers with twice the amount of stock, cutting the price of
algae production in half. This would increase profits which could then be used to further invest in
research and developing of algae biofuel production. If BP wishes to become our project’s primary
investor, the company will further strengthen its profound reputation at the forefront of alternative
energy and biofuel technologies.
In 8 months, we propose to engineer a strain of algae to double its oil content for an estimated cost
of $473,000. We will select a candidate alga and identify target DNA sequences involved in oil
production. Then using current genome engineering technology, we will modify and evolve it to
maximize oil content. After it is inspected for performance, the alga will be made available to the
BP Company as well as existing algae biofuel producers.
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SUPERIOR ALGAL OIL PRODUCTION THROUGH ACCELERATED
EVOLUTION
INTRODUCTION
PROBLEM STATEMENT:
The main obstacles involved in the economical production of algal biofuels are the energy requirement
of the extraction process and suboptimal oil production of the algae itself. Extracting oils from algae
requires a large amount of energy and algae are not optimally designed to produce oil.
BACKGROUND OF THE PROBLEM:
Biodiesel is produced from oils or fats derived from biomass, and is clean burning and renewable.
Currently many plants are grown to produce oils for biodiesel including soy, canola and palm. Algae
grow more quickly than other oil-producing crops and do not compete with food crops for valuable
agricultural land. Algae have high oil content, 500 times more than soy. Algae are clearly the best source
of bio-oil, but the cost of extraction limits its economic viability.
Several methods are used to extract the oil from algae. In the Ultrasonic-assisted extraction, intense
sound waves are used to create alternating pressure cycles. During the low-pressure cycle, small
bubbles of vapor are created in the liquid. The bubbles then collapse violently during a high-pressure
cycle, creating high speed liquid jets. The resulting shear forces break the cell structure and release the
oil, which can be skimmed from the top of the vat. Another method, osmotic shock, is a sudden
reduction in osmotic pressure, similarly causing the algae cells in a solution to rupture. Both these
extraction processes yield 80% of the algae oils; however, they are energy intensive, resulting in high
production costs.
The remaining 20% of the algae’s oil is retained by drying the algae into what is known as dry cake. An
expeller press can be used to remove about 80% of the remaining oil content from the dry cake. After
the oil has been extracted using an expeller, solvents like hexane and supercritical carbon dioxide can be
mixed with the algae as final attempt to remove the oil content and is up to 95% effective. The
remaining pulp can be mixed with cyclohexane to extract the oil. The oil dissolves in the cyclohexane,
and the pulp is filtered out from the solution. The oil and cyclohexane are separated by means of
distillation. This is done in special glassware to reduce corrosion from the solvent. Unfortunately, many
of these solvents are hazardous to the environment and the human body, and require expensive safety
equipment which adds to the cost of production.
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Figure 1: Origin Oil’s Single Step Oil Extraction Method
Steps to mitigate these barriers have been established in Origin Oil’s Single Step Process. The Single
Step Process extracts separates oil from the water and biomass in one step. The process does
not use chemicals or heavy machinery and no initial dewatering is required, and separates the
oil, water and biomass in less than an hour. The company’s Quantum Fracturing™ technology
combines with electromagnetic pulses and pH modification to break down cell walls and
release oil from the algae cells. Unfortunately, this technology is fairly new, and has only
progressed to the pilot stage.
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PERSONNEL INFORMATION:
GREGORY ROWE – The product of 4 billion years of evolution, Dr. Rowe is an alumnus of the
University of Washington Electrical Engineering department. He has 6 years of programming
experience and 3 years of experience with advanced robotics. He also has 4 years of experience
with genetically engineering new strains of E. coli in a bioengineering research institution utilizing
the MAGE process. His most recent project was a directed evolution project conducted at Harvard
University.
JAMES SONG - An alumnus from the University of Washington Chemical Engineering department,
he earned a PhD in Chemical Engineering at the age of 8. He later earned a PhD in Plant Biology at
the age of 12. Dr. Song has 11 years of experience with process design and computational fuel
extraction methods. He also has 9 years of experience with algae biology field studies. His
extensive experience in the biofuel industry and algae biology should prove essential to the goals of
Algae Rules.
ERIC STEPHANSON – An alumnus from the University of Washington Electrical Engineering
department, he earned a PhD in Electrical Engineering at the age of 13. Dr. Stephanson currently
has 7 years of experience with electromagnetics. He also has 5 years of experience with testing
electromagnetic fields on cells. Dr. Stephanson amazingly created a tesla coil in his garage at the age
of 14.
JAMES ZAJAC – An alumnus from the University of Washington Mechanical Engineering
department, Dr. Zajac has 18 years of experience with DNA splicing and 5 years of experience with
current algae biofuel extraction methods. Dr. Zajac worked with Dr. Rowe in the same directed
evolution project at Harvard University. His direct experience with algae will prove useful for Team
Algae Rules.
PURPOSE:
Team Algae Rules believes that there is an alternative solution to the problem: modify the algae itself to
allow for a more cost efficient extraction.
SCOPE:
The proposed project will include building an evolution machine and genetic modification of a common
alga to increase its oil yield and reduce the cost of processing.
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FORMAT:
The major sections that follow are Technical information, Statement of work, Timeline, Personnel
information, and projected cost. Technical information will detail our method to create a new strain of
algae.
TECHNICAL INFORMATION:
We plan to increase bio-oil production using Multiplex Automated Genome Engineering (MAGE) to
introduce genetic changes to an alga and artificially evolve it to improve oil content. The new strain will
be selected to produce higher amounts of oil types useful for biofuel. Dunaliella tertiolecta is a marine
green flagellate widely used for biofuel production. It is simple to cultivate, does not form biofilms,
contains about 23% oil (dry weight) and grows quickly giving it a high CO2 absorption rate. These
properties make D. tertiolecta an excellent candidate for alteration. Our goal is to double yields of useful
oils for the same amount of energy.
To accomplish this feat we intend to genetically modify an alga to produce a new strain that will more
efficiently convert solar radiation into oils. Harris H. Wang, a graduate student in the lab of George
Church (Professor of Genetics, Harvard Medical School), developed MAGE to modify bacteria. Starting
with a strain of E. coli that made a small amount of lycopene (the pigment that makes tomatoes red) he
synthesized 50,000 strands of DNA that vary slightly from 24 genes that are involved in lycopene
production. Putting the DNA and the bacteria into the MAGE device (nicknamed the evolution machine)
and after 35 cycles, a strain was produced that made 5 times as much lycopene as the original bacteria.
While engineering E. coli to make more lycopene has been done before, he accomplished in 3 days
something that took the entire biosynthesis industry several years. To show his achievement was not a
coincidence, he has since repeated the feat with the dye indigo. MAGE is an incredibly powerful tool for
genetic modification, able to produce profound results in a drastically shorter time than conventional
methods.
MAGE involves constructing thousands of DNA sequences that contain variations in genes that are
involved in the target trait, attaching them to viral enzymes and placing them in solution with the cells
to be modified. The solution is then stimulated electrically to open pores in the cell membrane, a
process called electroporation, allowing the modified DNA to infiltrate and incorporate itself in to the
bacterial genome. Cells can be repeatedly modified and evolved, leaving all the hard work of genetic
engineering to natural processes. A device to perform MAGE will cost around $70,000.
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PROJECT WORK SUMMARY
Project Timeline
Phase 1
7/24/11 9/12/11 11/1/11 12/21/11 2/9/12
3/30/12
Build evolution machine
Find suitable algae
Phase 2
Identify DNA sequence
Use MAGE process
Test and repeat
Production/Shipping
Figure 2: Project Timeline
The project will be conducted in two phases. The first phase consists of building the MAGE
evolution machine, finding a suitable algae strain to genetically alter, and identify the genes
involved in oil production. The second phase will encompass carrying out the MAGE process to
develop the proposed super-strain, running experiments to verify the performance of the strain,
and making it available to BP and other interested algae biofuel producers.
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COST INFORMATION
The itemized budget for our proposal is below. The overall cost of $473,000 is well within the
amount BP budgets for renewable energy research each year.
Table 1: Proposed itemized Budget
Evolution Machine
DNA synthesizer
Micro Bioreactors
Electroporator
Alga growth medium
Automation electrical/
mechanical
Maintenance
Estimated cost
$40,000
$10,000
$7,000
$1,000
$10,000
$73,000
$5,000
Salaries
Total Labor (~8
months)
$400,000
Total
$473,000
SUMMARY AND CONCLUSION
With more than 30 years combined experience in biosynthesis, the task of building a evolution
machine and altering an alga is within our capabilities. Our new alga will be simple to cultivate,
grown in existing bioreactors, and will not require any special equipment to process beyond
conventional extraction methods. This project represents a onetime cost; furthermore the MAGE
evolution machine can be used in the future to implement modifications to further reduce the cost
of biofuel production, such as weakening the cell wall to decrease the energy needed to rupture the
membrane.
We are requesting $473000 to develop a new strain of algae specifically for biofuel production. This
new alga will not only improve the profitability of biofuel production, it will also strengthen
domestic and international energy sectors, while improving Energy security for the future and
reducing dependence on fossil fuels.
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APPENDICES
REFERENCES
Kabellos, Michael (2009, February 3). greentechmedia
Algae Biodiesel: It’s $33 a Gallon Retrieved July 21, 2011,
from http://www.greentechmedia.com/articles/read/algae-biodiesel-its-33-a-gallon-5652/
Wang, Harris H.; Isaacs, Farren J.; Carr, Peter A.; Sun, Zachary Z.; Xu, George; Forest, Craig R.;
Church, George M. (2009, July 26) Nature
Programming cells by multiplex genome engineering and accelerated evolution
Retrieved July 17, 2011 from
http://www.nature.com/nature/journal/v460/n7257/full/nature08187.html
Wikipedia (June 4, 2011) Dunaliella
Retrieved July 19, 2011 from http://en.wikipedia.org/wiki/Dunaliella
United Nations ESCAP,
Regional Forum on Bioenergy Sector Development: Challenges, Opportunities, and Way Forward
Retrieved July 21, 2011 from http://www.unapcaem.org/publication/bioenergy.pdf
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