David Glass EUEC Presentation 02 02 10

Prospects for the Commercial Use of Genetic Engineering in Biofuel Production
David J. Glass, Ph.D.
D. Glass Associates, Inc.
Presented at EUEC 2010, Phoenix, AZ
February 2, 2010

Overview: Prospects for Use of Advanced Biotechnology for Biofuel Production
Summary of technology strategies.
Companies applying advanced biotechnology to biofuels.
Impact of biotechnology regulations on biofuels.
Commercialization status and prospects.

Technology Strategies

Biotechnologies Applicable to Biofuels
Classical microbial mutation and selection
Plant breeding
Recombinant DNA
Directed evolution
DNA shuffling
Metabolic engineering
Synthetic biology

Genetic Engineering Strategies: Microorganisms
Overexpress desired enzymes (e.g. to improve ability to process or degrade cellulosic feedstocks).
Engineer microorganisms to manufacture novel or improved industrial enzymes.
Create novel or synthetic microorganisms or new metabolic pathways to enable production of renewable fuels.

Genetic Engineering Strategies: Algae
Enhance algal growth rate.
Enhance or alter lipid biosynthesis.
Enhance photosynthesis.
Enable use of alternate food sources.
Enable secretion of lipids to aid oil/water separation.

Genetic Engineering Strategies: Plants
Overexpress cell-wall hydrolysis enzymes.
Cellulases, Hemicellulases, Ligninases.
Increase plant biomass, cellulosic biomass.
Cellulose biosynthetic enzymes.
Lignin modification to reduce need for pretreatment.
Down-regulate lignin biosynthesis.
Regulated gene expression, so traits are active only when needed.

Companies applying advanced biotechnology to biofuels

Companies Using or Developing Modified Microorganisms: Ethanol
BioEnergy International
DuPont Danisco
Glycos Biotechnologies
GreenTech America
Joule Biotechnologies
Mascoma
Microbiogen (non-GMO)
Promethegen
Qteros
TMO Renewables
Verenium
Zymetis
Note: Several of these companies are also developing other fuels.

Companies Using or Developing Modified Microorganisms: Biodiesel, Other Fuels
Biodiesel, petroleum fuels
Amyris Biotechnologies
Codexis
Glycos Biotechnologies
Joule Biotechnologies
LS9
Promethegen
Sequesco
Synthetic Genomics
Butanol, Isobutanol
Bioenergy International
Butamax (DuPont/BP)
Cobalt Biofuels
Gevo
Green Biologics, Ltd.
METabolic Explorer
TetraVitae Bioscience

Companies Using Newer Microbial Technologies
Amyris Biotechnologies: synthetic biology
Codexis: DNA shuffling
Glycos Biotechnologies: metabolic engineering
Joule Biotechnologies: synthetic biology
LS9: synthetic biology
Synthetic Genomics: synthetic biology
TMO Renewables: thermophilic microorganisms
Verdezyne: synthetic biology, metabolic engineering

Companies Manufacturing Enzymes for Biofuels using Modified Organisms
Microorganisms
AB Enzymes
Danisco/Genencor
Royal DSM N.V.
Dyadic International
Iogen Corporation
Novozymes A/S
Protéus
Plants
Infinite Enzymes (transgenic plants)
Medicago (transient expression)
Syngenta (transgenic plants)

Companies Reported to be Developing Genetically Modified Algae for Biofuels
Algenol Biofuels
Aurora Biofuels
AXI LLC
Global Green Solutions
Sapphire Energy
Solazyme
Synthetic Genomics
Targeted Growth
Note: companies included based on press, Internet reports: use of GMOs not necessarily confirmed on company websites. Not all these companies are necessarily using genetic engineering.

Companies Developing Transgenic Plants for Biofuels
Conventional Feedstocks
Agrivida: corn
ArborGen: purpose-growntrees
Edenspace Systems: corn
Targeted Growth: corn
Note: some of these companiesare also developing newer feedstocks.

Newer Feedstocks
Agragen: Camelina
Agrisoma: BrassicaandJatropha
Agrivida: switchgrass, sugarcane, sorghum, others
Ceres: non-food grasses
Edenspace Systems: switchgrass
Evogene: canola, soybean, others

Newer Feedstocks
Farmacule BioIndustries: sugarcane, tobacco
Mendel Biotechnology: grasses, others
Metabolix: switchgrass, oil crops, others
SG Biofuels: Jatropha
Targeted Growth: Camelina, canola, others

Impact of Biotechnology Regulations on Biofuels

Overview of U.S. Regulation of Environmental Biotechnology
Environmental Protection Agency
Microbial pesticides.
Plant pesticides (e.g. transgenic plants).
Engineered microorganisms not regulated by other agencies (e.g. nitrogen fixation, bioremediation, industrial applications).
U.S. Department of Agriculture
Transgenic plants, including plants producing industrial or pharmaceutical products.
Transgenic animals.

EPA Regulation of Engineered Microorganisms
EPA regulations under the Toxic Substances Control Act (TSCA) cover industrial uses of microorganisms not regulated by other agencies.
Applies to “intergeneric” organisms for industrial enzyme production, biotreatment, other bioprocessing uses.
Applicability to algae not certain.
Submit Microbial Commercial Activity Notice (MCAN) with data package 90 days before starting commercial use.
Most research and pilot projects are not regulated.
Some exemptions available for commercial use.

USDA Regulation of Transgenic Plants
Regulations issued in 1987 cover outdoor uses of most transgenic plants.
Rules have been continually relaxed, most agricultural uses require only agency notification.
Starting in 2003, permits now required for industrial, pharmaceutical, phytoremediation applications.
Permit applications: must be submitted 120 days in advance of proposed field use.
Commercialization via “delisting” petition process.
USDA currently considering revisions to regulations.

Regulation of Engineered Algae
Jurisdiction of EPA, USDA under biotech regulations unclear.
Proposed use of engineered algae in pharmaceutical production in Hawaii in 2005 generated controversy, no federal agency claimed jurisdiction: state law and state courts ultimately determined outcome.
2008 USDA opinion letter to Coastal BioMarine stated no USDA oversight over engineered algal strain if no “plant pest” sequences, and no oversight for smaller-scale use in contained reactors.
Reactor design may be critical in determining regulatory status.

Impact of Biotechnology Regulations
Would not necessarily affect all biofuel organisms, exemptions may be available.
For those projects affected, some additional activities and expense may be required.
Straightforward path to commercial approval, with many prior successes in agricultural, industrial biotechnology.
The regulatory process can be successfully managed.

Commercialization Status and Prospects

Commercial Status of Modified Microorganisms in Biofuels
Verenium: only MCAN reviewed by EPA under TSCA for modified ethanol production strain.
Demonstration or pilot plants in operation or under construction:
Ethanol production: BioEnergy, DuPont Danisco, Iogen, Mascoma, Qteros, Verenium.
Gevo: demonstration butanol plant.
Amyris: pilot plant for petroleum-equivalent fuels.
Pilot or commercial algae plants: Algenol, Aurora, Sapphire, Solazyme, Solix (not necessarily using engineered algae).

Field Uses of Transgenic Plants for Biofuels
Syngenta: Approval for commercial use of amylase-expressing corn in several countries, approval pending in U.S. since 2005.
Agrisoma: field trials in Canada of engineered Brassica and soybean, improved in oil quality, content and seed size for biodiesel use, 2009.
Infinite Enzymes, Edenspace, Targeted Growth, ArborGen: field trials in the U.S. of various species, including tobacco, corn, Eucalyptus, expressing biodegradative enzymes, 2006-09.

Notable Business Deals Involving Advanced Biotechnology for Biofuels
BP/Martek: Conversion of sugars to biodiesel.
BP/Verenium: Cellulosic biofuel development.
Chevron/Solazyme: Biodiesel production from algae.
Dow Chemical/Algenol: Pilot plant for algal production of ethanol.
ExxonMobil/Synthetic Genomics: Synthetic algal strains for biofuel production.
Royal Dutch Shell/Codexis: Novel methods for conversion of non-food biomass to biofuels.
Royal Dutch Shell/HR Petroleum: marine algae for biofuels.

Prospects for the Future: Commercialization Challenges
Use of modified organisms will face the same challenges as other biofuel applications:
Scale-up to commercial scale.
Process economics.
Competition from other biological or physical methods of biofuel manufacture.
Feedstock availability and cost; conflict with food use of crops
Availability of sufficient land to grow feedstocks or to build reactors.

Prospects for the Future: Commercialization Challenges
But use of modified organisms will also face unique challenges:
Technical hurdles to accomplish performance goals.
Recovering the high R&D costs of advanced biotech.
Government policies, including biotechnology regulation, especially outside the U.S.
Public acceptance of biotechnology solutions.

Prospects for the Future: Commercialization Strategies
Integration of biological processes with other downstream process improvements.
Diversification:
Augment energy production by co-production of value-added end-products: specialty chemicals, bioplastics.
Entry of tree, plant genetics companies into biofuels.
Carbon capture: design systems to capture CO2 streams for use as biofuel feedstock.
Novel feedstocks: use of cellulosic waste, novel nonfood crop species as biofuel feedstocks.

Thank you very much
David J. Glass, Ph.D.
D. Glass Associates, Inc.
124 Bird Street
Needham, MA 02492
Phone 617-653-9945
DGlassAssc@aol.com
www.dglassassociates.com
More detailed information on the subjects presented in this talk can be found at my blog:
http://dglassassociates.wordpress.com

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David Glass EUEC Presentation 02 02 10

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