1.
David J. Glass, Ph.D.
D. Glass Associates, Inc.
June 18, 2014

2.
Overview, Key Points
 History and overview of U.S. biotechnology
regulation affecting industrial uses of GM algae.
 Although the U.S. biotechnology framework singles out
modified organisms for additional scrutiny, approvals for
commercial uses of GM algae can be obtained with proper
planning.
 Scientific issues relating to regulation and risk
assessment of uses of GM algae.
 Legitimate scientific issues should be addressed in a risk
assessments for industrial uses of GM algae.
 Industry, government and academia can collaborate to
create research database to support risk assessments.
D. Glass Associates, Inc.

3.
Advanced Biotechnology can
Improve Usefulness of Algae
 Enhance algal growth rate.
 Enhance or alter lipid biosynthesis.
 Enhance photosynthesis.
 Enable use of alternate food sources.
 Create new biosynthetic pathways;
enable production of new products.
 Enable secretion of lipids
to aid oil/water separation.
Rosenberg et al. 2008, Li et al. 2008;
Angermayr et al. 2009, Radakovits et al 2010.
D. Glass Associates, Inc.

4.
U.S. Biotechnology Regulation:
Historical Perspective
 Biotechnology framework arose in the mid-1980s amid
considerable public interest and debate.
 The Reagan Administration decided against a
biotechnology-specific law, in favor of relying on existing
laws and regulations.
 Although meant to be “product-based” rather than
“process-based”, inevitably some regulations single out
uses of engineered organisms for greater scrutiny than
uses of unmodified organisms.
 Although broad enough to cover all commercial uses, the
greatest focus was on “deliberate releases” to the
environment.
D. Glass Associates, Inc.

5.
Overview of U.S. Biotechnology
Regulation
Environmental Protection Agency
 Microbial pesticides, plant pesticides.
 Engineered microorganisms used for other industrial
purposes.
U.S. Department of Agriculture
 Transgenic plants, potential plant pests.
 Plant-produced industrial products.
Food and Drug Administration
 Foods, food additives, pharmaceuticals.
TSCA biotech rule
USDA biotech rule
Product –specific regulation
Possible applicability to algae
D. Glass Associates, Inc.

6.
Regulatory Jurisdiction for
Engineered Algae
 Proposed use of engineered algae by Mera
Pharmaceuticals in pharmaceutical production in Hawaii
in 2005: 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.
 Can USDA, EPA work together on GM algae regulation,
with one agency designated as the lead?
D. Glass Associates, Inc.

7.
EPA TSCA Biotechnology
Regulations (1)
 Regulations adopted in 1997 under the Toxic Substances
Control Act (TSCA) cover commercial uses of “new
microorganisms”.
 Regulations cover only those industrial uses not regulated
by other agencies as foods, drugs, cosmetics, pesticides.
 Among covered activities: industrial enzyme production,
bioremediation, biotreatment, manufacture of fuels,
chemicals.
 New microorganisms are defined as “intergeneric”:
containing deliberate combinations of coding nucleic acids
from more than one taxonomic genus.
D. Glass Associates, Inc.

8.
EPA TSCA Biotechnology
Regulations (2)
EPA considers scope of coverage to include algae.
From the biotechnology rule:
“Microorganism” means an organism classified, using the 5-kingdom
classification system of Whittacker, in the kingdoms Monera (or
Procaryotae), Protista, Fungi, and the Chlorophyta and the
Rhodophyta of the Plantae, and a virus or virus-like particle.
From the rule’s preamble:
Therefore, this definition includes, but is not limited to, bacteria,
protozoa, fungi, mycoplasmas, mycoplasma-like organisms,
spiroplasmas, microphytoplanktons, green and red algae, viruses,
and virus-like particles (e.g., viroids, satellites, and virusoids).
D. Glass Associates, Inc.

9.
EPA TSCA Biotechnology
Regulations (3)
 Commercial use or importation of intergeneric organism
requires 90 day advance notification to EPA, through
submission of a Microbial Commercial Activity Notice
(MCAN).
 Most research and pilot projects would not require EPA review
if conducted in suitably “contained” facilities, with procedures
for controlled access, inactivation of wastes, emission controls,
worker notification.
 R&D with intergeneric organisms under non-contained
conditions, such as open-pond algae reactors, would require
EPA review through submission of a TSCA Experimental
Release Application (TERA) 60 days in advance of proposed
activity.
D. Glass Associates, Inc.

10.
EPA TSCA Biotechnology Rule:
TSCA Experimental Release Applications
 TERAs submitted to EPA 60 days in advance, describing
the organism, the proposed research, and the proposed
controls and monitoring procedures.
 EPA can approve or deny TERAs, or approve testing with
limitations or required monitoring.
 To date, 30 TERAs submitted, most for agricultural or
bioremediation microorganisms. All but three of these
have been approved.
 The 5 TERAs most recently filed and approved were from
Sapphire Energy, Inc., for open-pond research at UCSD
using different modified strains of the photosynthetic
green algae Scenedesmus dimorphus.
D. Glass Associates, Inc.

11.
EPA TSCA Biotechnology Rule: Microbial
Commercial Activity Notifications (MCANs)
 MCAN reporting required at least 90 days before
commencing commercialization or importing a “new
microorganism” for a TSCA purpose.
 MCAN requires submission of data to EPA.
 Microorganism identity, construction and its properties
 Potential health and environmental impacts.
 Information about the industrial process,
control/containment measures, worker exposure, possible
environmental release.
 EPA review, clearance of MCAN authorizes commercial
use for any purpose.
D. Glass Associates, Inc.

12.
EPA TSCA Biotechnology Rule:
Biofuel, Bio-Based Chemical MCANs
 Approx. 65 MCANs filed since 1997. Number and
frequency have increased in last 3 years.
 Most early MCANs covered GMOs for production of
industrial enzymes.
 14 MCANs for S. cerevisiae, 3 for Zymomonas
mobilis, all for ethanol production.
 Joule, Algenol have filed MCANs for cyanobacteria;
Solazyme has filed 2 MCANs for modified microalgae.
 Complete list available at
www.epa.gov/biotech_rule/pubs/submiss.htm.
D. Glass Associates, Inc.

13.
0
2
4
6
8
10
12
14
1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014
(to
date)
MCANs Submitted to EPA by Fiscal Year
D. Glass Associates, Inc.

14.
©2013 Joule. Rights Reserved. Proprietary & Confidential. www.jouleunlimited.com
Joule SunSpringsTM Hobbs: Where the vision becomes reality
Production Unit
Central Plant
 Commissioned in September 2012 to
test and demonstrate the platform at
increasingly larger scale
 Core production unit and
infrastructure in place to advance to
full process demonstration
 Achieving progress on numerous
variables, e.g. productivity, process
efficiency
 Sustained Sunflow®-E ethanol
production, with other products to
follow
 Facility and processes will become the
blueprint for future commercial plants
Biocatalyst Prep
©2013 Joule. Rights Reserved.

15.
Joule Unlimited: MCAN for
Modified Synechococcus
 First MCAN for commercial ethanol production strain filed
July 2012. The MCAN included all available information
to enable a risk assessment for the strain.
 EPA completed its review Fall 2012, concluded that the
use of the strain at Hobbs facility is unlikely to present
unreasonable risk.
 Consent Order signed July 2013, allows use of the strain
at Hobbs under specified conditions, with certain
additional testing, monitoring and reporting required.
 Additional MCANs will be needed for improvements to
the strain.
©2013 Joule. Rights Reserved.D. Glass Associates, Inc.

16.
EPA TSCA Biotechnology Rule:
Summary
 Algae and cyanobacteria covered under the rule;
EPA has recent experience with MCANs and TERAs
for algae and cyanobacteria.
 MCAN reviews for contained manufacturing have
been straightforward; EPA has recent track record
reviewing microorganisms for use in biofuels, bio-
based chemical production.
 The TERA process provides a stepwise approach to
regulation and risk assessment of outdoor uses.
D. Glass Associates, Inc.

17.
USDA Biotechnology
Regulations (1)
 Regulations issued in 1987, administered by USDA
Animal and Plant Health Inspection Service (APHIS).
 Regulations cover environmental uses or interstate
movement of organisms considered to be “potential plant
pests”.
 Potential plant pest status is based on presence of DNA
sequences from listed potential plant pests. Inclusion of
Agrobacterium on the list has captured most transgenic
plants.
 Most submissions have been for transgenic plants, some
for modified agricultural microorganisms.
D. Glass Associates, Inc.

18.
USDA Biotechnology
Regulations (2)
 Rules have generally not covered “contained” use of
organisms in manufacturing, but could cover open-
pond uses.
 Applicability to algae under current definitions not
clear; most algae strains would not be covered
unless engineered to contain DNA from potential
plant pest.
D. Glass Associates, Inc.

19.
USDA Biotechnology Regulation:
Oversight over R&D
 R&D use covered only if in open environment.
 Notifications: 30 day advance notice.
 Permits: submit application 120 days in advance.
 Permits generally required for industrial uses.
 Submissions require:
 Description of host organism and genetic
modifications.
 Description of field test; proposed procedures and
controls.
 Assess environmental impact of field use.
D. Glass Associates, Inc.

20.
USDA Biotechnology Regulation:
Approvals for Commercial Use
 Approval for commercial use and sale through
“petitions for nonregulated status”.
 Since early 1990s, USDA has approved over
100 petitions to allow commercial sale of
engineered crop plants.
 Commercial approvals can take 1-4 years,
and require USDA to prepare Environmental
Impact Statements.
D. Glass Associates, Inc.

21.
USDA Biotechnology Regulation:
Summary
 Tens of thousands of transgenic plant field tests
approved under these regulations, over 100
modified crop varieties approved for commercial
sale – no approvals for algae?
 Limited experience with R&D permits for novel
biofuel feedstocks, no commercial approvals.
 USDA likely to have scientific expertise to review
biology of photosynthetic algae, but applicability
under existing definitions unclear.
D. Glass Associates, Inc.

22.
International Biotechnology
Regulation
 Most countries around the world have biotechnology
regulations, in most cases based on principles of the
Cartagena Protocol on Biosafety.
 These countries differ from the U.S. in having unified
laws and regulations that cover all genetically
modified organisms.
 However, principles of risk assessments for algae
and microorganisms should be similar throughout
the world.
D. Glass Associates, Inc.

23.
Scientific Issues in the Regulation
of GM Algae (1)
Legitimate scientific issues need to be addressed
for many projects involving modified algal strains.
 Stability of vector and introduced genes.
 Possible deleterious functions encoded by
transgene(s) such as algal toxins.
 Potential for horizontal gene transfer, crossing to wild
algae species.
D. Glass Associates, Inc.

24.
Scientific Issues in the Regulation
of GM Algae (2)
 Potential for engineered strain to be transported
outside facility, survive and compete in environment.
 Potential persistence in the environment: soil or
water in vicinity of site of use.
 Potential disruption of natural ecosystems or native
algae populations.
 Creation or enhancement of harmful algal blooms or
ecologically disruptive algal blooms.
D. Glass Associates, Inc.

25.
Research Needs for GM Algae Risk
Assessment (contained research)
 Basic research into biology, ecology, natural history
of commercially-relevant wild type algae strains.
 Basic research addressing key regulatory concerns:
gene transmissibility, survival, persistence in
environment, genetics of algal toxin production.
 Laboratory, microcosm and macrocosm studies to
model behavior of modified species in the
environment.
D. Glass Associates, Inc.

26.
Research Needs for GM Algae Risk
Assessment (controlled field tests)
 Stepwise field studies with GM strains, carried out
under regulatory auspices, e.g. EPA TERA process.
 Conduct and monitor small-scale field tests of
modified algae in open ponds: develop database
addressing dispersal, environmental persistence, etc.
 Create a role for USDA and academic investigators in
EPA review of commercial uses of GM algae.
 Government support for field research: utilize DOE
national labs, other government facilities as field test
sites?
D. Glass Associates, Inc.

27.
Impact of Biotechnology
Regulations
 Efforts to promote interagency cooperation and the
participation of the research community, are important
to ensure a science-based regulatory framework.
 The existing biotech regulatory framework provides a
straightforward path to commercial approval, with
many prior successes in agricultural, industrial
biotechnology.
 The regulatory process can
be successfully managed
with proper advance planning.
D. Glass Associates, Inc.

28.
Thank you very much
David J. Glass, Ph.D.
D. Glass Associates, Inc.
124 Bird Street
Needham, MA 02492
Phone 617-653-9945
dglass@dglassassociates.com
www.dglassassociates.com
D. Glass Associates, Inc.