[Federal Register Volume 64, Number 101 (Wednesday, May 26, 1999)]
[Rules and Regulations]
[Pages 28358-28362]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 99-13151]
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DEPARTMENT OF HEALTH AND HUMAN SERVICES
Food and Drug Administration
21 CFR Part 184
[Docket No. 79G-0372]
Direct Food Substances Affirmed as Generally Recognized as Safe:
Cellulase Enzyme Preparation Derived From Trichoderma Longibrachiatum
for Use In Processing Food
AGENCY: Food and Drug Administration, HHS.
ACTION: Final rule.
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[[Page 28359]]
SUMMARY: The Food and Drug Administration (FDA) is amending its
regulations to affirm that cellulase enzyme preparation derived from
Trichoderma longibrachiatum (formerly called Trichoderma reesei) as
generally recognized as safe (GRAS) is for use in processing food. This
action is in response to a petition filed by the AAC Consulting Group,
Inc., on behalf of Novo Laboratories, Inc.
DATES: This regulation is effective May 26, 1999. The Director of the
Office of the Federal Register approves the incorporation by reference
in accordance with 5 U.S.C. 552(a) and 1 CFR part 51 of a certain
publication in Sec. 184.1250 (21 CFR 184.1250), effective May 26, 1999.
FOR FURTHER INFORMATION CONTACT: Nega Beru, Center for Food Safety and
Applied Nutrition (HFS-206), Food and Drug Administration, 200 C St.
SW., Washington, DC 20204, 202-418-3097.
SUPPLEMENTARY INFORMATION:
I. Background
In accordance with the procedures described in Sec. 170.35 (21 CFR
170.35), AAC Consulting Group, Inc. (formerly Arthur A. Checci, Inc.),
7445 Wisconsin Ave., suite 850, Bethesda MD 20814, on behalf of Novo
Nordisk BioChem North America, Inc. (formerly Novo Laboratories, Inc.),
State Rd. 1003, P.O. Box 576, Franklinton, NC 27525-0576, submitted a
petition (GRASP 9G0260) requesting affirmation that cellulase enzyme
preparation derived from a nonpathogenic strain of T. reesei (later
renamed T. longibrachiatum) used for processing food is GRAS.
Cellulase, the enzyme, is to be distinguished from cellulase enzyme
preparation, which contains cellulase as the principal active
component, but it also contains other components derived from the
production organism and fermentation media. This document will refer to
the former as ``cellulase'' and the latter as ``cellulase enzyme
preparation.''
In the Federal Register of November 27, 1979 (44 FR 67731), FDA
published a notice of filing of GRASP 9G0260, and gave interested
parties an opportunity to submit comments. FDA received one comment in
response to the notice. The comment urged the agency to affirm the GRAS
status of the cellulase enzyme preparation without restricting its use
in food other than to require that the use of the enzyme be consistent
with current good manufacturing practice.
II. Standards for GRAS Affirmation
Under Sec. 170.30 (21 CFR 170.30), general recognition of safety
may be based only on the views of experts qualified by scientific
training and experience to evaluate the safety of substances added to
food. The basis of such views may be either : (1) Scientific
procedures, or (2) in the case of a substance used in food prior to
January 1, 1958, experience based on common use in food
(Sec. 170.30(a)). General recognition of safety based upon scientific
procedures requires the same quantity and quality of scientific
evidence as is required to obtain approval of a food additive
regulation and ordinarily is based upon published studies, which may be
corroborated by unpublished studies and other data and information
(Sec. 170.30(b)). General recognition of safety through experience
based on common use in food prior to January 1, 1958, may be determined
without the quantity or quality of scientific evidence required for
approval of a food additive regulation and ordinarily is based upon
generally available data and information.
In its petition, Novo Nordisk BioChem North America, Inc., relied
on scientific procedures, primarily published studies, scientific
papers and books, to demonstrate the safety and identity of the
cellulase enzyme and the production strain from which it is derived.
The petitioner provided published studies documenting that cellulase
enzyme preparation derived from nontoxicogenic, nonpathogenic T.
longibrachiatum is GRAS.
In evaluating this petition, the agency reviewed information
concerning: (1) The production organism, (2) the identity and function
of the cellulase enzyme, (3) the production and purification of the
cellulase enzyme preparation, (4) the use of the cellulase enzyme
preparation in the production of food products, and (5) the safety of
the enzyme preparation.
III. Safety Evaluation
A. Introduction
Commercial enzyme preparations that are used in food processing
typically are not chemically pure, but they contain, in addition to the
enzyme component, other components that derive from the production
organism and fermentation media, residual amounts of processing aids,
and substances used as stabilizers, preservatives or diluents. Issues
relevant to a safety evaluation of the enzyme preparation therefore
include the safety of the enzyme component, the safety of the enzyme
source, and the safety of processing aids and other substances added
during the manufacturing process. A safety evaluation of an enzyme
preparation also includes consideration of dietary exposure to that
preparation.
B. Production Organism
In a submission dated December 7, 1988, the petitioner informed
the agency that the International Commission on Taxonomy of Fungi
(ICTF) had decided to rename the source organism, a fungus known for
its high cellulase productivity, from T. reesei, to T. longibrachiatum
(Ref. 1). The petitioner presented published studies to assess
potential pathogenicity of T. longibrachiatum in mice, rabbits, and
guinea pigs (Ref. 2). No adverse reactions were reported in these
studies. The petitioner also included in its petition the results of a
search of several scientific data bases including Biological Abstracts,
1977-83; Chemical Abstracts, 1977-83; Scisearch, 1978-83; Medline,
1980-83; and Food Science and Technology Abstracts, 1969-83. The
petitioner states that these searches demonstrate that T.
longibrachiatum is well known and available to the scientific
community, and the data bases contain studies in which the
microorganism, or enzymes derived from it, were utilized without any
evidence of pathogenicity or toxicogenicity being associated with their
use. The searches did not identify a single report that T.
longibrachiatum is the etiological agent of a disease in man or
animals. The agency concludes, based upon the published information
presented in the petition (Refs. 2 through 6) that the production
organism T. longibrachiatum has been adequately identified and
determined to be nontoxicogenic and nonpathogenic (Ref. 7).
C. Identity and Function of the Cellulase Enzyme
Cellulase is the accepted name for the enzyme that catalyzes the
endohydrolysis of 1,4-beta-glucosidic linkages in cellulose (Ref. 8).
The enzyme will also hydrolyze 1,4-linkages in beta-glucans. The
enzymatically formed reaction products are mainly glucose and
cellobiose, a disaccharide composed of two glucose molecules. According
to the recommendations of the International Union of Pure and Applied
Chemistry and the International Union of Biochemistry (1972), cellulase
has the following designation: Cellulase, E.C. 3.2.1.4 (Ref. 9). FDA
concludes that generally available and accepted data and information
establish that the cellulase that is the subject of this document is
capable of achieving its intended technical effect.
[[Page 28360]]
D. Production of Cellulase Enzyme Preparation
The production process for cellulase enzyme preparation from T.
longibrachiatum is described in GRASP 9G0260 and can be summarized as
follows. A pure culture of T. longibrachiatum is aseptically grown in a
typical culture medium such as one containing potato starch, soybean
meal, corn steep liquor, or dextrose. Mineral salts, such as phosphates
and sulfates, are included in the medium which also contains an
antifoaming agent and a surfactant. The fermentation is conducted at 26
to 32 deg.C with aeration and maximal agitation. Cell growth and the
possible presence of foreign microorganisms are monitored by taking
samples before inoculation of the fermenter, every 24 hours during
cultivation, and before transfer/harvesting.
After 100 to 170 hours, the culture broth is subjected to
flocculation and filtration. The enzyme, which is secreted into the
extracellular medium, is separated from the mycelium by action of the
flocculating agent. This material is then removed by filtration using a
filter aid. The enzyme, which remains in solution, is concentrated by
ultrafiltration or vacuum evaporation at 30 to 40 deg.C. The enzyme
suspension is then dried to a powder by spray drying or concentrated in
liquid form by vacuum evaporation. The packaged finished product,
powder or liquid, is shipped or stored at 4 deg.C.
The agency finds that the fermentation generating organism is
maintained in a manner to avoid contamination and genetic changes, that
the fermentation is a pure culture fermentation initially and is
monitored for purity periodically during the culture period, and that
the filtration step in the purification process would remove any viable
production organisms from the final product (Ref. 7). The agency
further finds that, because the potential impurities in the cellulase
enzyme preparation that may originate from the source or manufacturing
process do not raise any basis for concern about the safe use of the
preparation, the general requirements for enzyme preparations as
described in the ``Food Chemicals Codex,'' 4th ed. (1996) (Ref. 10),
which are being incorporated by reference in new Sec. 184.1250 in
accordance with 5 U.S.C. 552(a) and 1 CFR part 51, are adequate as
minimum criteria for food-grade cellulase enzyme preparation.
E. Use in Food
The function of cellulase enzyme preparation in food production
includes uses such as the breakdown of the cellulose in citrus
products, removal of fiber from edible oil press cakes, increase in
starch recovery from potatoes, extraction of proteins from leaves and
grasses, tenderizing fruits and vegetables prior to cooking, extraction
of essential oils and flavoring material from plant materials, the
preparation of animal feeds, and other uses that are discussed in
publications such as the Handbook of Food Additives (Refs. 11 and 12) .
The petitioner also presented additional published information
that the cellulase enzyme preparation performed its intended technical
effect in the production of various food materials. Cellulase enzyme
preparation has been shown to be effective in the degradation of
vegetable tissues and in the extraction of green tea components,
vegetable proteins and starches. Cellulase enzyme preparation is also
capable of modifying food materials such as vegetables, rice, and
soybeans to markedly influence the digestibility, cooking quality,
shape, and the yield of nutrients (Ref. 13).
The agency has considered the estimated dietary exposure to
cellulase enzyme preparation from its proposed use (Refs. 14 and 15).
Enzymes, including the petitioned cellulase, are used in small
quantities in food to accomplish their intended effects. In addition,
many food processes that use cellulase also include removal of
insoluble solids, a processing step that should remove most of the
added enzyme preparation. Nonetheless, in calculating the estimated
dietary exposure to cellulase enzyme preparation, the agency made the
conservative assumptions that no cellulase enzyme preparation is
removed from the food by processing, and all foods that may be treated
with cellulase enzyme preparation will be so treated. The agency
concludes that the dietary exposure to cellulase enzyme preparation
does not present a basis for concern about the safety of its use (Refs.
16 and 17).
F. Safety Studies
The petitioner has provided published studies with the cellulase
enzyme preparation, corroborated with unpublished studies, to
demonstrate that the enzyme preparation is safe for use in food. The
petitioner provided published oral acute toxicity studies with mice,
rats, and dogs and oral subchronic studies with rats and dogs (Ref. 2).
No significant adverse effects were noted in these studies.
A published toxicity study with in utero exposure, and a
teratogenicity study, both conducted with rats, reported no adverse
effects at levels up to 5 percent in the diet (Ref. 2). The petitioner
also provided published mutagenicity studies involving the Ames test,
chromosomal aberration tests, and dominant lethal tests (Ref. 2). There
was no evidence of mutagenicity of the cellulase enzyme preparation in
any of these tests. Other published studies with the cellulase enzyme
preparation provided by the petitioner include an inhalation study in
rats; skin and eye irritation tests in rabbits; a skin irritation test
in humans; and a skin sensitivity test in guinea pigs and humans.
Finally, because certain species of Trichoderma are known to produce
substances that inhibit the growth of microorganisms, the petitioner
tested the culture broth of T. longibrachiatum for antibiotics or
toxins; the tests were negative (Ref. 2).
The agency has reviewed the published safety studies in the
petition along with other available information. The agency concludes
that the published safety data support the use of cellulase enzyme
preparation from T. longibrachiatum for the enzymatic breakdown of
cellulose in processing food (Refs. 16 and 17).
IV. Conclusions
The agency has evaluated all available information and finds,
based upon the published information about the identity and function of
cellulase, that the enzyme component of cellulase enzyme preparation
will achieve its intended technical effect and raises no toxicity
concerns. The agency further finds, based upon generally available and
accepted information, that when the cellulase enzyme preparation is
manufactured in accordance with Sec. 184.1250, the source, T.
longibrachiatum, and the manufacturing process will not introduce
impurities into the preparation that may render its use unsafe. Finally
the agency finds that dietary exposure to the cellulase enzyme
preparation from the petitioned use does not present a basis for
concern about the safe use of the cellulase enzyme preparation.
Therefore, the agency concludes, based on the evaluation of published
data and information, and based upon scientific procedures
(Sec. 170.30(b)), that use of the cellulase enzyme preparation derived
from T. longibrachiatum for the enzymatic breakdown of cellulose in
processing food is GRAS. Therefore, the agency is affirming that the
use of
[[Page 28361]]
cellulase enzyme preparation from T. longibrachiatum described in the
regulation set out below is GRAS (21 CFR 184.1(b)(1)) with no
limitations other that current good manufacturing practice.
V. Environmental Effects
The agency has carefully considered the potential environmental
effects of this action. FDA has concluded that the action will not have
a significant impact on the human environment, and that an
environmental impact statement is not required. The agency's findings
of no significant impact and the evidence supporting these findings,
contained in an environmental assessment, may be seen in the Dockets
Management Branch (HFA-305), Food and Drug Administration, rm. 1061,
Rockville, MD 20852, between 9 a.m. and 4 p.m., Monday through Friday.
VI. Analysis of Impacts
A. Analysis for Executive Order 12866
FDA has examined the impacts of this final rule under Executive
Order 12866. Executive Order 12866 directs agencies to assess the costs
and benefits of available regulatory alternatives and, when regulation
is necessary, to select regulatory approaches that maximize net
benefits (including potential economic, environmental, public health
and safety effects; distributive impacts; and equity). According to
Executive Order 12866, a regulatory action is significant if it meets
any one of a number of specified conditions, including having an annual
effect on the economy of $100 million, adversely affecting in a
material way a sector of the economy, competition, or jobs, or if it
raises novel legal or policy issues. FDA finds that this final rule is
not a significant regulatory action as defined by Executive Order
12866. In addition, it has been determined that this final rule is not
a major rule for the purpose of congressional review.
The primary benefit of this action is to remove uncertainty about
the regulatory status of the petitioned substance. No compliance costs
are associated with this final rule because no new activity is required
and no current or future activity is prohibited by this rule.
B. Regulatory Flexibility Analysis
FDA has examined the impacts of this final rule under the
Regulatory Flexibility Act. The Regulatory Flexibility Act (5 U.S.C.
601-612) requires Federal agencies to consider alternatives that would
minimize the economic impact of their regulations on small entities. In
compliance with the Regulatory Flexibility Act, FDA finds that this
final rule will not have a significant impact on a substantial number
of small entities.
No compliance costs are associated with this final rule because no
new activity is required and no current or future activity is
prohibited. Accordingly, under the Regulatory Flexibility Act (5 U.S.C.
605(b)), the agency certifies that this final rule will not have a
significant economic impact on a substantial number of small entities.
VII. Paperwork Reduction Act of 1995
This final rule contains no collections of information. Therefore,
clearance by the Office of Management and Budget under the Paperwork
Reduction Act of 1995 is not required.
VIII. Effective Date
As this rule recognizes an exemption from the food additive
definition in the Federal Food, Drug, and Cosmetic Act, and from the
approval requirements applicable to food additives, no delay in
effective date is required by the Administrative Procedure Act (5
U.S.C. 553(d)). The rule will therefore be effective immediately (5
U.S.C. 553(d)(1)).
IX. References
The following references have been placed on display in the
Dockets Management Branch (address above) and may be seen by interested
persons between 9 a.m. and 4 p.m., Monday through Friday.
1. Cannon, P. F., ``International Commission on the Taxonomy of
Fungi (ICTF): Name Changes in Fungi of Microbiological Industrial
and Medical Importance. Part 2,'' Microbiological Sciences, 3(9):
285 to 287, 1986.
2. Hjortkjaer, R. K. et al., ``Safety Evaluation of Celluclast
TM, an Acid Cellulase Derived From Trichoderma reesei,'' Food
Chemical Toxicity, 21 (1) 55 to 63, 1986.
3. ``Specifications for the Identity and Purity of Some Enzymes
and Certain Other Substances'' in the Fifteenth Report of the Joint
FAO/WHO Expert Committee on Food Additives; WHO Food Additive
Series, No. 2, pp. 3 to 5, 1972 .
4. Rifai, M. A., ``A Revision of the Genus Trichoderma'' in
Mycological Papers, No. 116, pp. 42 to 44, 1969.
5. Simmons, E. G., ``Classification of Some Cellulase Producing
Trichoderma Species'' in Second International Mycological Congress,
Book of Abstracts, p. 618, 1977.
6. Church, B. D., N. A. Nash, and W. Brosz, ``Use of Fungi
Imperfecti in Treating Food Processing Wastes,'' in Development in
Industrial Microbiology, vol. 13, pp. 30 to 46, 1972.
7. Memorandum from Food and Cosmetics Microbiology Branch, FDA,
to GRAS review Branch, FDA, January 10, 1980.
8. King, K. W., and M. I. Vassal, ``Enzymes of the Cellulase
Complex,'' in Cellulase and Their Applications, edited by R. F.
Gould ``Advances in Chemistry Series'' #95, American Chemical
Society, Washington, DC, pp. 7 to 25, 1969.
9.``Enzyme Nomenclature,'' recommendations (1972) of the
International Union of Pure and Applied Chemistry and the
International Union of Biochemistry, American Elsevier, New York,
pp. 212 to 213, 1975.
10. ``Monograph on Enzyme Preparations'' in Food Chemicals
Codex, 4th ed., National Academy Press, Washington, DC, pp. 129 to
134, 1996.
11. Underkofler, L. A., ``Enzymes'' in CRC Handbook of Food
Additives, edited by T. E. Furia, The Chemical Rubber Co., Ohio, pp.
80 to 82, 1968.
12. Malmos, H., ``Industrial Applications of Cellulase: Enzyme
Applications in Food, Pharmaceuticals and Other Industries,'' in
American Institute of Chemical Engineers Symposium Series, vol. 74,
1976/77, pp. 93-99.
13. Toyoma, N., ``Applications of Cellulases in Japan,'' pp.
359 to 390, in Cellulases and Their Applications, edited by R. F.
Gould, Advances in Chemistry Series 195, American Chemical Society,
Washington, DC, 1969.
14. Memorandum from Food and Color Additives Review Section,
FDA, to Direct Additives Branch, FDA, February 21, 1989.
15. Memorandum from Food and Color Additives Review Section,
FDA, to Direct Additives Branch, FDA, June 22, 1990.
16. Memorandum from Additives Evaluation Branch, FDA, to Direct
Additives Branch, FDA, July 11, 1990.
17. Memorandum from Additives Evaluation Branch, FDA, to Direct
Additives Branch, FDA, June 29, 1993.
List of Subjects in 21 CFR Part 184
Food ingredients, Incorporation by reference.
Therefore, under the Federal Food, Drug, and Cosmetic Act and
under authority delegated to the Commissioner of Food and Drugs and
redelegated to the Director, Center for Food Safety and Applied
Nutrition, 21 CFR part 184 is amended as follows:
PART 184--DIRECT FOOD SUBSTANCE AFFIRMED AS GENERALLY RECOGNIZED AS
SAFE
1. The authority citation for 21 CFR part 184 continues to read as
follows:
Authority: 21 U.S.C. 321, 342, 348, 371.
2. Section 184.1250 is added to subpart B to read as follows:
Sec. 184.1250 Cellulase enzyme preparation derived from Trichoderma
longibrachiatum.
(a) Cellulase enzyme preparation is derived from a nonpathogenic,
nontoxicogenic strain of Trichoderma
[[Page 28362]]
longibrachiatum (formerly T. reesei). The enzyme, cellulase, catalyzes
the endohydrolysis of 1,4-beta-glycosidic linkages in cellulose. It is
obtained from the culture filtrate resulting from a pure culture
fermentation process.
(b) The ingredient meets the general and additional requirements
for enzyme preparations in the monograph specifications on enzyme
preparations in the ``Food Chemicals Codex,'' 4th ed. (1996), pp. 129
to 134, which is incorporated by reference in accordance with 5 U.S.C.
552(a) and 1 CFR part 51. Copies are available from the National
Academy Press, 2101 Constitution Ave. NW., Box 285, Washington, DC
20055 (Internet ``http://www.nap.edu''), or may be examined at the
Center for Food Safety and Applied Nutrition's Library, 200 C St. SW.,
rm. 3321, Washington, DC, or at the Office of the Federal Register, 800
North Capitol St. NW., suite 700, Washington, DC.
(c) In accordance with Sec. 184.1(b)(1), the ingredient is used in
food with no limitation other than current good manufacturing practice.
The affirmation of this ingredient as generally recognized as safe
(GRAS) as a direct human food ingredient is based upon the following
current good manufacturing practice conditions of use:
(1) The ingredient is used in food as an enzyme as defined in
Sec. 170.3(o)(9) of this chapter for the breakdown of cellulose.
(2) The ingredient is used in food at levels not to exceed current
good manufacturing practice.
Dated: May 17, 1999.
L. Robert Lake,
Director, Office of Policy, Planning and Strategic Initiatives, Center
for Food Safety and Applied Nutrition.
[FR Doc. 99-13151 Filed 5-25-99; 8:45 am]
BILLING CODE 4160-01-F