Impact of Government Biotechnology Regulations on Biofuel Development David J. Glass, Ph.D., D. Glass Associates, Inc. Companies are using genetic engineering to improve biofuels microorganisms by... Data requirements under TSCA Description of the recipient and new microorganism. Genetic construction of the new microorganism. Phenotype and ecological characteristics of the new microorganism. Production process, byproducts. Worker exposure and environmental release. Health and environmental effects data. D. Glass Associates, Inc. Experience with Biotech Regulations BioTechnica International , EPA approval, field tests of nitrogen fixing microorganisms, 1988-90, USDA approval, transgenic plant tests, 1989-90. Strategies to use transgenic plants for biofuels applications include ... Engineered microorganisms might be subject to EPA regulation under the Toxic Substances Control Act (TSCA) Dozens of microbial strains have been reviewed and approved under TSCA Transgenic plants are regulated by the U.S. Department of Agriculture (USDA) Scientific issues under USDA regulations Currently advising companies developing improved microorganisms, transgenic plants, for biofuel uses. Applied PhytoGenetics , USDA permits for phytoremediation with transgenic plants, 2003-05. Envirogen , favorable EPA ruling on bioreactor containment, 1992. Energy Biosystems, favorable EPA ruling on bioreactor containment, 1995. Applied PhytoGenetics 2003 phytoremediation field test, Danbury, Connecticut. D. Glass Associates, Inc. D. Glass Associates, Inc. is a consulting company specializing in regulatory affairs, licensing and technology transfer, and strategic patent management in several fields of biotechnology. David Glass, Ph.D. has over 25 years experience in the biotechnology industry, including extensive experience with the regulation of industrial and agricultural uses of engineered organisms, and 17 years experience in technology licensing. David J. Glass, Ph.D. D. Glass Associates, Inc. 124 Bird Street Needham, MA 02492 Phone (617) 653-9945 Fax (781) 449-8045 [email_address] www.dglassassociates.com Overexpressing desired enzymes (e.g. to improve ethanol fermentation). Engineering microbes to manufacture novel or improved industrial enzymes. Creating novel or synthetic microorganisms to produce biologically-based fuels resembling petroleum fuels. Before adoption of 1997 regulations, EPA approved applications for 32 microbial strains used in contained manufacturing and 11 strains for outdoor field testing. Since 1997, EPA has reviewed 17 notifications for use of engineered microbes for enzyme manufacture; all have been cleared for commercial use. Since 1997, EPA has approved18 applications for outdoor field testing of engineered microorganisms, including 6 for nitrogen-fixing bacteria and 12 for environmental remediation or monitoring. One strain of engineered nitrogen-fixing bacteria approved for commercial sale under TSCA. Regulations issued in 1987 (7 CFR Part 340) cover outdoor uses of most transgenic plants. Rules have been continually relaxed, most agricultural uses no longer need permits, require only agency notification. Starting in 2003, permits now required for industrial, pharmaceutical, phyto- remediation applications. Permit applications: must be submitted 120 days in advance. Introducing genes encoding key biodegradative enzymes, to enhance the conversion of cellulose into fermentable sugars. Improving agronomic traits to enhance crop yield (e.g. pest resistance). Stability of vector and introduced genes. Presence of deleterious functions encoded by transgene. Reproduction and pollen/seed dispersal mechanisms. Ability to outcross with related species (particularly wild relatives). Potential weediness (ability to compete, survive and spread in the environment). Physical isolation from sexually compatible species. Regulations adopted in 1997 (40 CFR Part 725) cover industrial uses of microorganisms not regulated by other agencies. Applies to “intergeneric” organisms for industrial enzyme production, biotreatment, other bioprocessing uses. Notify EPA, submit data package 90 days before starting commercial use. Most research and pilot projects are not regulated. These approvals include over 60 permits for industrial and pharmaceutical applications since 2004. USDA has also approved 73 transgenic crop varieties for commercial use. Recommended strategies for regulatory approval Designate one company official as the project manager. Early contact with regulatory agency: 6-12 months before planned start date. Presubmission meeting to discuss project and data requirements. Follow-up correspondence to resolve any unanswered questions. Company commitment to devote appropriate resources to obtain data requested by agency. When submitting application, allow extra time beyond statutory timeframes. BioTechnica Rhizobium field test, approved under TSCA, 1988. Envirogen reactor for TCE biotreatment. Use of advanced biotechnology strategies in biofuels projects may be affected by biotechnology regulations of the U.S. EPA or the U.S. Department of Agriculture. With advanced planning, it should be possible to successfully manage a program to obtain timely regulatory approvals under the biotechnology rules. USDA has approved over 13,000 field tests of transgenic plants since 1987

Data requirements under TSCA

Description of the recipient and new microorganism.

Genetic construction of the new microorganism.

Phenotype and ecological characteristics of the new microorganism.

Production process, byproducts.

Worker exposure and environmental release.

Health and environmental effects data.

Overexpressing desired enzymes (e.g. to improve ethanol fermentation).

Engineering microbes to manufacture novel or improved industrial enzymes.

Creating novel or synthetic microorganisms to produce biologically-based fuels resembling petroleum fuels.

Before adoption of 1997 regulations, EPA approved applications for 32 microbial strains used in contained manufacturing and 11 strains for outdoor field testing.

Since 1997, EPA has reviewed 17 notifications for use of engineered microbes for enzyme manufacture; all have been cleared for commercial use.

Since 1997, EPA has approved18 applications for outdoor field testing of engineered microorganisms, including 6 for nitrogen-fixing bacteria and 12 for environmental remediation or monitoring.

One strain of engineered nitrogen-fixing bacteria approved for commercial sale under TSCA.

Regulations issued in 1987 (7 CFR Part 340) cover outdoor uses of most transgenic plants.

Rules have been continually relaxed, most agricultural uses no longer need permits, require only agency notification.

Starting in 2003, permits now required for industrial, pharmaceutical, phyto- remediation applications.

Permit applications: must be submitted 120 days in advance.

Introducing genes encoding key biodegradative enzymes, to enhance the conversion of cellulose into fermentable sugars.

Improving agronomic traits to enhance crop yield (e.g. pest resistance).

Stability of vector and introduced genes.

Presence of deleterious functions encoded by transgene.

Reproduction and pollen/seed dispersal mechanisms.

Ability to outcross with related species (particularly wild relatives).

Potential weediness (ability to compete, survive and spread in the environment).

Physical isolation from sexually compatible species.

Regulations adopted in 1997 (40 CFR Part 725) cover industrial uses of microorganisms not regulated by other agencies.

Applies to “intergeneric” organisms for industrial enzyme production, biotreatment, other bioprocessing uses.

Notify EPA, submit data package 90 days before starting commercial use.

Most research and pilot projects are not regulated.

Designate one company official as the project manager.

Early contact with regulatory agency: 6-12 months before planned start date.

Presubmission meeting to discuss project and data requirements.

Follow-up correspondence to resolve any unanswered questions.

Company commitment to devote appropriate resources to obtain data requested by agency.

When submitting application, allow extra time beyond statutory timeframes.