Document ID: EPA-HQ-OPP-2006-0642-0017
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2007-04-18T04:00Z

Glyphosate Oxidoreductase and the Genetic Material Necessary for Its 

Production in All Plants; Exemption From Tolerance Requirement On All
Raw 

Agricultural Commodities 

[Federal Register: October 8, 1997 (Volume 62, Number 195)]

[Rules and Regulations]

[Page 52505-52509]

From the Federal Register Online via GPO Access [wais.access.gpo.gov]

[DOCID:fr08oc97-12]

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ENVIRONMENTAL PROTECTION AGENCY

40 CFR Part 180

[OPP-300552; FRL-5745-2]

RIN 2070-AB78

Glyphosate Oxidoreductase and the Genetic Material Necessary for

Its Production in All Plants; Exemption From Tolerance Requirement On

All Raw Agricultural Commodities

AGENCY:  Environmental Protection Agency (EPA).

ACTION:  Final rule.

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SUMMARY:  This document establishes an exemption from the requirement

of a tolerance for residues of the plant-pesticide inert ingredients

glyphosate oxidoreductase (GOX) and the genetic material necessary for

its production in all plants when used as plant-pesticides in or on all

raw agricultural commodities (RACs). Monsanto Company submitted a

petition to EPA under the Federal Food, Drug, and Cosmetic Act (FFDCA)

as amended by the Food Quality Protection Act of l996 (FQPA) requesting

the exemption from the requirement of a tolerance. This regulation

eliminates the need to establish a maximum permissible level for

residues of this plant-pesticides in or on all RACS.

DATES:  This regulation is effective on October 8, 1997. Written

objections and requests for hearings must be received by EPA on or

before December 8, 1997.

ADDRESSES:  Written objections and hearing requests, identified by the

docket control number OPP-300552, may be submitted to: Hearing Clerk

(1900), Environmental Protection Agency, Rm. M3708, 401 M St., SW.,

Washington, DC 20460. Fees accompanying objections and hearing requests

shall be labeled ``Tolerance Petition Fees'' and forwarded to: EPA

Headquarters Accounting Operations Branch, OPP (Tolerance Fees), P.O.

Box 360277M, Pittsburgh, PA 15251. A copy of any objections and hearing

requests filed with the Hearing Clerk should be identified by the

docket control number OPP-300552 and submitted to: Public Information

and Records Integrity Branch, Information Resources and Services

Division (7506C), Office of Pesticide Programs, Environmental

Protection Agency, 401 M St., SW., Washington, DC 20460. In person,

bring a copy of objections and hearing requests to: Rm. 1132, Crystal

Mall #2, 1921 Jefferson Davis Highway, Arlington, VA.

     A copy of objections and hearing requests filed with the Hearing

Clerk may also be submitted electronically by sending electronic mail

(e-mail) to:

    opp-docket@epamail.epa.gov.

    Copies of objections and hearing requests must be submitted as an

ASCII file avoiding the use of special characters and any form of

encryption. Copies of objections and hearing requests will also be

accepted on disks in WordPerfect in 5.1 file format or ASCII file

format. All copies of objections and hearing requests in electronic

form must be identified by the docket control number OPP-300552. No

Confidential Business Information (CBI) should be submitted through e-

mail. Additional information on CBI can be found in VII. of this

document. Electronic copies of objections and hearing requests on this

rule may be filed online at many Federal Depository Libraries.

Additional information on electronic submissions can be found in Unit

VIII. of this document.

FOR FURTHER INFORMATION CONTACT:  By mail: Mike Mendelsohn,

Biopesticides and Pollution Prevention Division, Office of Pesticide

Programs, Environmental Protection Agency, 401 M St., SW., Washington,

DC 20460. Office location, telephone number, and

[[Page 52506]]

e-mail address: 5th Floor Crystal Station, 2800 Crystal Drive,

Arlington, VA , (703) 308-8715); e-mail:
mendelsohn.mike@epamail.epa.gov.

SUPPLEMENTARY INFORMATION:  In the Federal Register of January 24, 1997

(62 FR 3682) (FRL-5380-2), EPA issued a notice pursuant to section

408(d) of FFDCA, 21 U.S.C. 346a(d) announcing the filing of a pesticide

petition for an exemption from the requirement for a tolerance by

Monsanto Company, 700 Chesterfield Parkway, North St. Louis, MO 63198.

The notice contained a summary of the petition prepared by the

petitioner and this summary contained conclusions and arguments to

support its conclusion that the petition complied with the FQPA (Pub.

L. 104-170). The petition requested that an exemption from the

requirement of a tolerance be established for the plant-pesticides GOX

and the genetic material necessary for its production in plants in or

on all RACS. There were no comments or requests for referral to an

advisory committee received in response to the notice of filing. The

data submitted in the petition and other relevant material have been

evaluated. The toxicology and other data listed below were considered

in support of this exemption from the requirement of a tolerance.

I. Risk Assessment and Statutory Findings

    New section 408(c)(2)(A)(I) of FFDCA allows EPA to establish an

exemption from the requirement of a tolerance (the legal limit for a

pesticide chemical residue in or on a food) only if EPA determines that

the tolerance is ``safe'' to mean that ``there is a reasonable

certainty that no harm will result from aggregate exposure to the

pesticide chemical residue, including all anticipated dietary exposures

and all other exposures for which there is reliable information.'' This

includes exposure through drinking water and in residential settings,

but does not include occupational exposure. Section 408(c)(2)(B) of

FFDCA requires EPA to give special consideration to exposure of infants

and children to the pesticide chemical residue in establishing a

tolerance and to ``ensure that there is a reasonable certainty that no

harm will result to infants and children from aggregate exposure to the

pesticide chemical residue...'' EPA performs a number of analyses to

determine the risks from aggregate exposure to pesticide residues.

First, EPA determines the toxicity of pesticides. Second, EPA examines

exposure to the pesticide through food, drinking water, and through

other exposures that occur as a result of pesticide use in residential

settings.

II. Aggregate Risk Assessment and Determination of Safety

    Consistent with section 408(b)(2)(D) of FFDCA, EPA has reviewed the

available scientific data and other relevant information in support of

this action and considered its validity, completeness, and reliablitiy

and the relationship of this information to human risk. EPA has

sufficient data to assess the hazards of glyphosate oxidoreductase

(GOX), and to make a determination on aggregate exposure, consistent

with section 408(c)(2) of FFDCA, for an exemption from the requirement

of a tolerance for residues GOX in or on all RACS. EPA's assessment of

the dietary exposures and risks associated with establishing the

exemption follows.

A. Toxicological Profile

    Glyphosate oxidoreductase (GOX) catalyzes the conversion of

glyphosate to aminomethylphosphonic acid (AMPA) and glyoxylate in a 1:1

stoichiometry while consuming \1/2\ mole of oxygen as a cosubstrate.

GOX requires flavin adenine dinucleotide (FAD) and magnesium for

activity; therefore, it is more appropriately designated an apoenzyme.

    The gene for Gox was originally isolated from Achromobacter sp.

Strain LBAA. The GOX protein was then sequenced and the gene was

synthesized with an added signal sequence and the codons modified in

the guanine and cytosine nucleic acid (GC) content to yield higher

plant expression. Two modified GOX proteins are specified in this rule.

They are designated GOX and GOXv247. Both versions have an identifier

of ``(M4-C1)'' in the data submissions which indicates that the protein

was expressed in E. coli for testing purposes. The GOX protein retains

the same amino acid sequence as the native protein and has additional

four amino acid sequence N-terminus (reminanats of an added signal

sequence). In GOXv247, the gene sequence of the native protein was

altered resulting in changes to three amino acids in the sequence of

the resulting protein along with the remains of the added signal

sequence mentioned previously. These changes did not negatively affect

the enzymatic activity of either protein.

    The GOX variants GOX and GOXv247, expressed in E. coli and

originating from the synthetic GOX gene optimized for protein

expression in plants, showed similarity to the native GOX protein when

expressed in E. coli. These similarities are seen as comparable

molecular weights, immunoreactivity, amino acid sequence and enzymatic

activity.

    The data submitted regarding potential health effects of GOX and

GOXv247 includes information on the characterization of the expression

of GOX in corn, the acute oral toxicity of GOX and GOXv247, and in

vitro digestibility studies of the proteins. The applicability of the

results of these studies to evaluate human risk and the validity,

completeness, and reliability of the available data from the studies

were considered.

    Both variants of the GOX protein (GOX and GOXv247) are rapidly

degraded in simulated gastric fluid (GF) and simulated intestinal fluid

(IF). After a fifteen-second incubation in GF, both variants have less

than 90% of their initial protein epitopes by western blot analysis.

Enzyme activity loss is also greater than 90% in both GOX variants when

assayed after a 1-minute incubation in GF. Similar results are seen in

simulated IF. Western blot assays show that both variants are greater

than 90% degraded by 30-second incubation in IF. However, the enzyme

activity assays show that the GOX activity lasts longer in IF than

variant GOXv247. After a 10-minute IF incubation, the activity

decreased to about 48% of initial for GOX whereas GOXv247 was already

greater than 90% inactive.

    Two findings, found in the in vitro digestibility studies, that are

remarkable are: GOXv247 displays a more rapid degradation in the IF

compared to unaltered GOX, apparently due to the single amino acid

substitutions; and antibody recognition is lost prior to a significant

loss of enzyme activity indicating that western blots may not always

accurately track functional protein degradation.

    None of the amino acid sequences of known allergens or proteins

involved in coeliac disease were shown to have similarity to the GOX

protein as defined by eight identical and contiguous amino acids in a

sequence. However, the assertion that a lack of allergenicity can be

established by comparison of sequences to known allergens is

questionable. While this is the best approximation at present, there is

no scientific basis to assume that the presence of eight contiguous and

homologous amino acids in a protein will predict its allergenicity. The

assumption is based on the finding that the presence of an eight amino

acid sequence in one allergen was associated with the epitope

responsible for IgE recognition. Alteration of this sequence

[[Page 52507]]

reduced IgE binding and hence allergenicity. The converse experiment,

to introduce the sequence into a non-allergenic protein and create an

allergen, has not been attempted experimentally.

    The acute oral toxicity test of bacterially-derived GOX and GOXv247

proteins showed no test substance related deaths at doses of 91.3

milligrams per kilogram (mg/kg) and 104 g/kg respectively.

Expression data on the GOX protein expressed in corn grains ranges from

undetectable levels to a high of 11.70 micro grams per gram (mg/g)

freshweight. This indicates that it would require 8,547 kg corn grain

per kg bodyweight to receive the 100 mg/kg dose that was administered

to the mice.

    However, residue chemistry data were not required for a human

health effects assessment of the subject plant-pesticide inert

ingredients because of the lack of mammalian toxicity. Both available

information concerning the dietary consumption patterns of consumers

(and major identifiable subgroups of consumers including infants and

children) and safety factors which, in the opinion of experts qualified

by scientific training and experience to evaluate the safety of food

additives, are generally recognized as appropriate for the use of

animal experimentation data were not evaluated because the lack of

mammalian toxicity at high levels of exposure demonstrate the safety of

the product at levels above possible maximum exposure levels. This is

similar to the Agency position regarding toxicity and the requirement

of residue data for the microbial Bacillus thuringiensis. [See 40 CFR

158.740(b).] For microbial products, further toxicity testing to verify

the observed effects and clarify the source of the effects (Tiers II

and III) and residue data are triggered by significant acute effects in

studies such as the mouse oral toxicity study.

    The acute oral toxicity data submitted support the prediction that

the GOX proteins would be non-toxic to humans. When proteins are toxic,

they are known to act via acute mechanisms and at very low dose levels

[Sjoblad, Roy D., et al. ``Toxicological Considerations for Protein

Components of Biological Pesticide Products,'' Regulatory Toxicology

and Pharmacology 15, 3-9 (1992)]. Therefore, since no effects were

shown to be caused by the plant-pesticides, even at relatively high

dose levels, the GOX protein is not considered toxic.

    Adequate information was submitted to show that the GOX test

materials derived from microbial cultures was biochemically and,

functionally similar to the proteins produced by the plant-pesticide

inert ingredient in corn. Production of microbially produced protein

was chosen in order to obtain sufficient material for testing. In

addition, the in vitro digestibility studies indicate the proteins

would be rapidly degraded following ingestion.

    The genetic material necessary for the production of the plant-

pesticides active and inert ingredients are the nucleic acids (DNA)

which comprise genetic material encoding these proteins and) their

regulatory regions. ``Regulatory regions'' are the genetic material

that control the expression of the genetic material encoding the

proteins, such as promoters, terminators, and enhancers. DNA is common

to all forms of plant and animal life and the Agency knows of no

instance where these nucleic acids have been associated with toxic

effects related to their consumption as a component of food. These

ubiquitous nucleic acids as they appear in the subject plant-pesticide

inert ingredient have been adequately characterized by the applicant.

Therefore, no mammalian toxicity is anticipated from dietary exposure

to the genetic material necessary for the production of the subject

active and inert plant pesticidal ingredients.

B. Sensitivity of Subgroups

    The Agency has considered available information on the variability

of the sensitivities of major identifiable subgroups of consumers

including infants and children and the physiological differences

between infants and children and adults and effects of in utero

exposure to the plant-pesticides. Since GOX is a protein, allergenic

sensitivities were considered. Current scientific knowledge suggests

that common food allergens tend to be resistant to degradation by heat,

acid, and proteases, are glycosylated and are present at high

concentrations in the food. Data has been submitted which demonstrate

that the GOX proteins are rapidly degraded by gastric fluid in vitro

and are non-glycosylated. Thus, the potential for the GOX proteins to

be a food allergens is minimal.

C. Cumulative Effects

     The Agency has considered available information on the cumulative

effects of such residues and other substances that have a common mode

toxicity. These considerations included the cumulative effects on

infants and children of such residues and other substances with a

common mechanism of toxicity. Because there is no indication of

mammalian toxicity to these plant-pesticides, there are no cumulative

effects.

D. Aggregate Exposures

     The Agency has considered available information on the aggregate

exposure levels of consumers (and major identifiable subgroups of

consumers) to the pesticide chemical residue and to other related

substances. These considerations include dietary exposure under the

tolerance exemption and all other tolerances or exemptions in effect

for the plant-pesticides chemical residue, and exposure from non-

occupational sources. Exposure via the skin or inhalation is not likely

since the plant-pesticides are contained within plant cells which

essentially eliminates these exposure routes or reduces these exposure

routes to negligible. Oral exposure, at very low levels, may occur from

ingestion of processed food products and drinking water. However a lack

of mammalian toxicity and the digestibility of the plant-pesticides has

been demonstrated. Regarding exposure via residential or lawn use to

infants and children, the Agency concludes that such exposure would

present no risk due to the lack of toxicity.

    Section 408 of FFDCA provides that EPA shall apply an additional

10-fold margin of exposure (MOE) (safety) for infants and children in

the case of threshold effects to account for pre- and post-natal

toxicity and the completeness of the database unless EPA determines

that a different MOE (safety) will be safe for infants and children. In

this instance EPA believes there is reliable data to support the

conclusion that the plant-pesticides are not toxic to mammals,

including infants and children, and thus there are no threshold effects

of concern. As a result, the provision requiring an additional MOE does

not apply.

III. Endocrine Effects

    EPA does not have any information regarding endocrine effects for

these kinds of pesticides at this time. The Agency is not requiring

information on the endocrine effects of these plant-pesticides at this

time; and Congress allowed 3 years after August 3, 1996, for the Agency

to implement a screening and testing program with respect to endocrine

effects.

IV. Analytical Method

    The Agency is establishing an exemption from the requirement of a

tolerance without numerical limitation; therefore, it has concluded

that an analytical method is not required for enforcement purposes for

GOX and the genetic material necessary for their production.

[[Page 52508]]

V. Conclusion

     There is a reasonable certainty that no harm will result from

aggregate exposure to the U.S. population, including infants and

children, to the GOX protein and the genetic material necessary for

that production. This includes all anticipated dietary exposures and

all other exposures for which there is reliable information. The Agency

has arrived at this conclusion because, as discussed above, no toxicity

to mammals has been observed for the plant-pesticides. As a result, EPA

establishes an exemption from tolerance requirements pursuant to

section 408(j)(3) of FFDCA for GOX and the genetic material necessary

for their production in all plants.

     Glyphosate Oxidoreductase [GOX or GOXv247] and the genetic

material necessary for its production in all plants are exempt from the

requirement of a tolerance when used as plant-pesticide inert

ingredients in all plant RACs. ``Genetic material necessary for its

production'' means the genetic material which comprise genetic material

encoding the GOX proteins and their regulatory regions. ``Regulatory

regions'' are the genetic material that control the expression of the

genetic material encoding the GOX proteins, such as promoters,

terminators, and enhancers.

VI. Objections and Hearing Requests

     The new FFDCA section 408(g) provides essentially the same process

for persons to ``object'' to a regulation for an exemption from the

requirement of a tolerance issued by EPA under new section 408(d) of

FFDCA as was provided in the old section 408 and in section 409 of

FFDCA. However, the period for filing objections is 60 days, rather

than 30 days. EPA currently has procedural regulations which governs

the submission of objections and hearing requests. These regulations

will require some modification to reflect the new law. However, until

those modifications can be made, EPA will continue to use those

procedural regulations with appropriate adjustments to reflect the new

law.

    Any person may, by December 8, 1997 file written objections to any

aspect of this regulation and may also request a hearing on those

objections. Objections and hearing requests must be filed with the

Hearing Clerk, at the address given under the ``ADDRESSES'' section (40

CFR 178.20). A copy of the objections and/or hearing requests filed

with the Hearing Clerk should be submitted to the OPP docket for this

rulemaking. The objections submitted must specify the provisions of the

regulation deemed objectionable and the grounds for the objections (40

CFR 178.25). Each objection must be accompanied by the fee prescribed

by 40 CFR 180.33(i). If a hearing is requested, the objections must

include a statement of the factual issue(s) on which a hearing is

requested, the requestor's contentions on such issues, and a summary of

any evidence relied upon by the objector (40 CFR 178.27). A request for

a hearing will be granted if the Administrator determines that the

material submitted shows the following: There is a genuine and

substantial issue of fact; there is a reasonable possibility that

available evidence identified by the requestor would, if established,

resolve one or more of such issues in favor of the requestor, taking

into account uncontested claims or facts to the contrary; and

resolution of the factual issue(s) in the manner sought by the

requestor would be adequate to justify the action requested (40 CFR

178.32).

VII. Confidential Business Information

    Information submitted in connection with an objection or hearing

request may be claimed confidential by marking any part or all of that

information as CBI. Information so marked will not be disclosed except

in accordance with procedures set forth in 40 CFR part 2. A copy not

contain CBI must be submitted for inclusion in the public record.

Information not marked confidential may be disclosed publicly by EPA

without prior notice.

VIII. Public Record and Electronic Submissions

    EPA has established a record for this rulemaking under docket

control number OPP-300552 (including any comments and data submitted

electronically). A public version of this record, including printed,

paperversions of electronic comments, which does not in include any

information claimed as CBI, is available for inspection from 8:30 a.m.

to 4 p.m., Monday through Friday, excluding legal holidays. The public

record is located in Room 1132 of the Public Information and Records

Integrity Branch, Information Resources and Services Division (7506C),

Office of Pesticide Programs, Environmental Protection Agency, Crystal

Mall #2, 1921 Jefferson Davis Highway, Arlington, VA.

    Electronic comments may be sent directly to EPA at:

    opp-docket@epamail.epa.gov.

    Electronic comments must be submitted as an ASCII file avoiding the

use of special characters and any form of encryption.

     The official record for this rulemaking, as well as the public

version, as described above, will be kept in paper form. Accordingly,

EPA will transfer any copies of objections and hearing requests

received electronically into printed, paper form as they are received

and will place the paper copies in the official rulemaking record. The

official rulemaking record is the paper record maintained at the

Virginia address in ``ADDRESSES'' at the beginning of this document.

IX. Regulatory Assessment

    This final rule establishes an exemption from the tolerance

requirement under FFDCA section 408(d) in response to a petition

submitted to the Agency. The Office of Management and Budget (OMB) has

exempted these types of actions from review under Executive Order

12866, entitled Regulatory Planning and Review (58 FR 51735, October 4,

1993). This final rule does not contain any information collections

subject to OMB approval under the Paperwork Reduction Act (PRA),

44U.S.C. 3501 et seq., or impose any enforceable duty or contain any

unfunded mandate as described under Title II of the Unfunded Mandates

Reform Act of 1995 (UMRA) (P.L. 104-4). Nor does it require any prior

consultation as specified by Executive Order 12875, entitled Enhancing

the Intergovernmental Partnership (58 FR 58093, October 28, 1993), or

special considerations as required by Executive Order 12898, entitled

Federal Actions to Address Environmental Justice in Minority

Populations and Low-Income Populations (59 FR 7629, February 16, 1994),

or require OMB review in accordance with Executive Order 13045,

entitled Protection of Children from Environmental Health Risks and

Safety Risks (62 FR 19885, April 23, 1997).

    In addition, since tolerances and exemptions that are established

on the basis of a petition under FFDCA section 408(d), such as the

exemption in this final rule, do not require the issuance of a proposed

rule, the requirements of the Regulatory Flexibility Act (RFA) (5

U.S.C. 601 et seq.) do not apply. Nevertheless, the Agency previously

assessed whether establishing tolerances, exemptions from tolerances,

raising tolerance levels or expanding exemptions might adversely impact

small entities and concluded, as a generic matter, that there is no

adverse economic impact. The factual basis for the Agency's generic

certification for tolerance actions published on May 4, 1981 (46 FR

24950), and was provided

[[Page 52509]]

to the Chief Counsel for Advocacy of the Small Business Administration.

X. Submission to Congress and the General Accounting Office

    Under 5 U.S.C. 801(a)(1)(A), as added by the Small Business

Regulatory Enforcement Fairness Act of 1996, the Agency has submitted a

report containing this rule and other required information to the U.S.

Senate, the U.S. House of Representatives, and the Comptroller General

of the General Accounting Office prior to publication of this rule in

today's Federal Register. This is not a ``major rule'' as defined by 5

U.S.C. 804(2).

 List of Subjects in 40 CFR Part 180

     Environmental protection, Administrative practice and procedure,

Agricultural commodities, Pesticides and pests, Reporting and

recordkeeping requirements.

    Dated: September 25, 1997.

Stephen L. Johnson,

Acting Director, Office of Pesticide Programs.

    Therefore, 40 CFR chapter I is amended as follows:

PART 180--[AMENDED]

    1. The authority citation for part 180 continues to read as follows:

    Authority: 21 U.S.C. 346a and 371.

    2. Section 180.1190 is added to subpart D to read as follows:

Sec. 180.1190  Glyphosate Oxidoreductase [GOX or GOXv247] and the

genetic material necessary for its production in all plants; exemption

from the requirement of a tolerance.

     Glyphosate Oxidoreductase [GOX or GOXv247] and the genetic

material necessary for its production in all plants are exempt from the

requirement of a tolerance when used as plant-pesticide inert

ingredients in all plant RACs. Genetic material necessary for its

production means the genetic material which comprise genetic material

encoding the GOX proteins and their regulatory regions. Regulatory

regions are the genetic material that control the expression of the

genetic material encoding the GOX proteins, such as promoters,

terminators, and enhancers.

[FR Doc. 97-26190 Filed 10-7-97; 8:45 am]

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