[Federal Register Volume 63, Number 49 (Friday, March 13, 1998)]
[Proposed Rules]
[Pages 12421-12426]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 98-6571]
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DEPARTMENT OF HEALTH AND HUMAN SERVICES
Food and Drug Administration
21 CFR Part 184
[Docket No. 89G-0393]
Direct Food Substances Affirmed as Generally Recognized as Safe;
Egg White Lysozyme
AGENCY: Food and Drug Administration, HHS.
ACTION: Tentative final rule.
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SUMMARY: The Food and Drug Administration (FDA) is issuing a tentative
final rule to amend its regulations to affirm that egg white lysozyme
enzyme preparation, when labeled by the common or usual name ``egg
white lysozyme'' to identify its source, is generally recognized as
safe (GRAS) for use in preventing late blowing of cheese caused by the
bacterium Clostridium tyrobutyricum during cheese production. This
action is in response to a petition submitted by Fordras S.A. (formerly
SPA-Societa Prodotti Antibiotici S.p.A.). FDA has tentatively concluded
that this use of the egg white lysozyme enzyme preparation is GRAS only
when the ingredient statement for both bulk and packaged food that
contains cheese manufactured using egg white lysozyme includes the
common or usual name ``egg white lysozyme'' to identify the source of
the protein. To give interested persons an opportunity to comment on
this condition of use required for GRAS status, FDA is issuing this
tentative final rule.
DATES: Submit written comments by May 27, 1998.
ADDRESSES: Submit written comments to the Dockets Management Branch
(HFA-305), Food and Drug Administration, 12420 Parklawn Dr., rm. 1-23,
Rockville, MD 20857.
FOR FURTHER INFORMATION CONTACT: Linda S. Kahl, Center for Food Safety
and Applied Nutrition (HFS-206), Food and Drug Administration, 200 C
St. SW., Washington, DC 20204, 202-418-3101.
SUPPLEMENTARY INFORMATION:
I. Background
In accordance with the procedures described in Sec. 170.35 (21 CFR
170.35), SPA-Societa Prodotti Antibiotici S.p.A., now Fordras S.A.,
Milan, Italy, submitted a petition (GRASP 9G0355) requesting that egg
white lysozyme used to inhibit the bacterium C. tyrobutyricum to
prevent late blowing of cheese during production be affirmed as GRAS as
a direct human food ingredient. FDA published the notice of filing for
this petition in the Federal Register of October 27, 1989 (54 FR
43861), and gave interested persons until December 26, 1989, to submit
written comments.
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 directly
or indirectly 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, through experience based on common use in
food. 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 procedures required for approval of a food
additive regulation, but ordinarily is based upon generally available
data and information concerning the pre-1958 history of use of the
substance.
FDA has evaluated Fordras S.A.'s petition on the basis of
scientific procedures to whether the petitioned use of egg white
lysozyme enzyme preparation to prevent the late blowing of cheese
caused by the bacterium C. tyrobutyricum during cheese production is
GRAS. In evaluating the petition, FDA considered published and
unpublished data and information relating to the identity of,
characteristic properties of, and estimated dietary exposure to the
enzyme component (i.e., lysozyme) of the petitioned enzyme preparation
(Refs. 1 through 7). FDA also considered that the source of the
petitioned enzyme preparation, egg white, has been safely consumed by
humans as a source of food protein throughout recorded history, and,
therefore, is GRAS (Sec. 170.30(d)), and that the methods used for
extracting lysozyme from the egg white source do not ordinarily alter
the chemical identity and characteristic properties of enzymes (Ref.
8). FDA also considered published scientific review articles (Refs. 1
and 2) and a generally available trade association bulletin (Ref. 7)
discussing the use of egg white lysozyme enzyme preparation for its
[[Page 12422]]
technical effect of preventing late blowing of cheese contaminated with
C. tyrobutyricum as well as generally available information documenting
that this intended use of the petitioned enzyme preparation has been
approved in several European countries (Refs. 9 through 13). Finally,
FDA considered generally available and accepted information relating to
processing aids used in the manufacture of the enzyme preparation and
generally available and accepted specifications for food-grade enzyme
preparations (Ref. 14).
III. Safety Evaluation
When present as a contaminant in milk used for cheesemaking, the
pasteurization-resistant bacterium C. tyrobutyricum ferments lactate to
produce carbon dioxide, hydrogen, and volatile organic acids. This
fermentation causes a defect in cheese manufacture known as ``late
blowing,'' which is typified by abnormal levels of open texture
accompanied by undesirable odors and flavors. Late blowing can be a
serious economic problem in the manufacture of several varieties of
cheese (Refs. 1, 2, and 7).
The contamination by C. tyrobutyricum of milk used for
cheesemaking, although reducible by good husbandry and hygienic milking
practices, is unavoidable. Although treatment with certain chemical
agents has been shown to be effective against the problems raised by
this contamination, treatment with lysozyme enzyme preparation has been
found to be the most effective method of managing the late blowing of
cheese contaminated with C. tyrobutyricum (Refs. 1 and 2).
A. The Enzyme Component
Enzymes are proteins or conjugated proteins (i.e., a protein that
contains a nonamino acid moiety such as a carbohydrate) produced by
plants, animals, and microorganisms that function as biochemical
catalysts (American Heritage Dictionary of the English Language). Most
enzymes are very specific in their ability to catalyze only certain
chemical reactions; this high degree of specificity and strong
catalytic activity are the most important functional properties of
enzymes (Ref. 15).
The Commission on Enzymes of the International Union of
Biochemistry has devised a systematic strategy for naming enzymes. This
system combines a naming system and a numbering system. For most
enzymes, the systematic name is derived from the names of the
substrate, product, and type of reaction. The systematic number is
based on the class and subclasses to which the enzyme belongs. The
systematic name of lysozyme is peptidoglycan N-acetylmuramoylhydrolase.
Its systematic number is EC No. 3.2.1.17 and its Chemical Abstracts
Service Registry Number (CAS Reg. No.) is 9001-63-2.
Lysozyme was first discovered by A. Fleming, who identified
lysozyme as an antibacterial enzyme present in nasal mucus membrane
(Ref. 3). Subsequently, it was learned that the antibacterial activity
of lysozyme occurs because of its ability to catalyze the hydrolysis of
the structural polysaccharide peptidoglycan present in cell walls of
certain bacteria (Ref. 2). Lysozyme activity has been shown to be
present in bacteria, fungi, plants, and almost all animal tissues, with
the highest levels found in secretions (including milk, mucus, saliva,
and tears) and eggs. Lysozyme is believed to function in all of these
organisms and tissues as an endogenous antimicrobial substance (Refs. 1
and 2).
Lysozyme was the first enzyme to have the details of its three-
dimensional structure published (Ref. 4), and it has become one of the
best characterized of all enzymes, serving as an example for studies of
enzyme mechanism and molecular evolution (Refs. 5 and 6 ). Lysozymes
from various organisms are very similar to one another. Egg white
lysozyme differs very little in structure, amino acid sequence and
composition, catalytic mechanism, and substrate specificity from the
enzyme found in human milk, saliva, mucus, and tears (Refs. 3 and 6).
The petitioner provided two published scientific review articles
(Refs. 1 and 2) that discuss the use of egg white lysozyme in cheese
and other food. The petitioner also provided a generally available
trade association bulletin (Ref. 7) that focuses on the use of egg
white lysozyme for its technical effect of preventing late blowing in
cheese. This bulletin describes the late blowing defect and how it
arises, traditional chemical control measures (other than the use of
lysozyme) to reduce the problem, and the increasing interest in using
lysozyme as a replacement for traditional chemical control measures. In
addition, the petitioner provided generally available information
documenting that this intended use of the petitioned enzyme preparation
has been approved in several countries, including Denmark, France,
Germany, Italy, and Spain (Refs. 9 through 13).
FDA considered the estimated dietary exposure to lysozyme for the
proposed use in cheese (Refs. 16 and 17). Lysozyme accounts for
approximately 3.5 percent of the total protein of domestic hen egg
whites (Ref. 7). Whole eggs contain lysozyme at a level of
approximately 3,300 parts per million (ppm). The petitioner reported
that cheese manufactured using egg white lysozyme enzyme preparation
contains a maximum of 400 ppm of lysozyme, or at least 8 times less
than eggs on a weight basis. FDA has estimated a long-term mean intake
of lysozyme to be 74 milligrams per person per day (mg/p/d) for
consumers of eggs and 3.8 mg/p/d for consumers of cheese; the
respective 90th percentile intakes are estimated to be 163 mg/p/day and
8.1 mg/p/day. Egg whites from which lysozyme is extracted will be
subsequently consumed in other food uses. Thus, there will be no long-
term net increase in lysozyme intake by the general population because
egg whites without lysozyme will replace egg whites in current use that
contain lysozyme (Ref. 16). On a per eating occasion basis, lysozyme
intake for cheese consumers may be 16 mg on average, or 22 mg at the
90th percentile level. For comparison, a per eating occasion lysozyme
intake for egg consumers may be 264 mg on average, or 416 mg at the
90th percentile level. Thus, lysozyme intake per eating occasion due to
cheese consumption may constitute 5 to 6 percent of lysozyme intake due
to egg consumption (Ref. 17).
In general, issues relevant to a safety evaluation of proteins such
as the enzyme component of an enzyme preparation are potential toxicity
and allergenicity (Ref. 18). Proteins derived from egg whites do not
raise toxicity concerns because egg whites have been safely consumed by
humans as a source of food throughout recorded history without any
reports of toxicity. However, proteins derived from egg whites do raise
allergenicity concerns because, as with many common foods, there have
been reports that consumption of egg whites can cause an allergic
reaction in certain individuals, particularly children (Ref. 19).
Therefore, FDA considered the question of whether the lysozyme
component of egg whites is allergenic.
In evaluating this question, FDA considered a report of an in vitro
study of the binding of antibodies to specific egg proteins, where the
antibodies were derived from the serum of patients known to be allergic
to eggs (Ref. 20). This report suggests that lysozyme was an allergen
for some individuals who became sensitive to egg whites. Although this
study does not establish that ingestion of egg white lysozyme in cheese
will actually cause a clinically
[[Page 12423]]
significant allergic reaction in such sensitive individuals, FDA is not
aware of any data or information that would refute the study's
inference that egg white lysozyme may be allergenic. Accordingly, FDA
is proposing labeling, as discussed below, to alert the sensitive
population to the presence of egg white lysozyme in cheese.
A related question is whether egg white lysozyme, when present in
cheese, is capable of inducing an allergenic response in susceptible
individuals who have not previously consumed egg whites, e.g., because
their customary diet excludes eggs. This question is no different than
for any other food containing egg white when consumed by individuals
with unknown susceptibility to eggs. The proposed label declaration
would provide such individuals with the same protection as that
provided by other egg-containing products with ingredient labeling.
Thus, individuals who experience an allergic reaction to lysozyme-
containing cheese could identify egg white lysozyme as a possible cause
of the reaction.
B. Enzyme Source, Manufacturing Methods, and Processing Aids
Commercial preparations of lysozyme are derived from domestic hen
egg whites using ion exchange methods and selective precipitation to
isolate a highly purified protein fraction that contains mainly
lysozyme but also may contain small amounts of other egg white
proteins. Consistent with the agency's finding in its GRAS affirmation
of microparticulated protein product (55 FR 6384, February 23, 1990),
FDA finds that egg whites have been safely consumed by humans
throughout recorded history and, therefore, are GRAS (Sec. 170.30(d)).
The agency evaluated the methods used to isolate the enzyme lysozyme
from egg whites. These methods are based on generally available and
accepted principles of protein purification (Ref. 8). Such methods, if
appropriately selected, do not ordinarily alter the chemical identity
and characteristic properties of enzymes. Therefore, these methods do
not materially change the quality, utility, functionality, or safety of
enzymes. Moreover, the retention of the antibacterial activity that is
characteristic of egg white lysozyme when egg white-derived lysozyme
enzyme preparation is used in cheese evidences that lysozyme in the
manufactured enzyme preparation remains unaltered from the lysozyme in
egg whites. This is corroborative evidence of the fact that the methods
used to isolate lysozyme from egg whites do not materially change the
quality, utility, functionality or safety of the enzyme lysozyme.
Enzyme preparations used in food processing are usually not
chemically pure but contain, in addition to the enzyme component,
materials that derive from the enzyme source. As mentioned above, egg
white lysozyme enzyme preparation may contain small amounts of other
egg white proteins. A related question is whether such proteins that
may be present in the enzyme preparation are allergenic. Even if
present, other source-derived proteins would not be a concern because
the proposed label declaration for egg white lysozyme would alert
individuals who are sensitive to egg whites to the possible presence of
other proteins derived from egg whites.
In addition to source-derived materials, enzyme preparations used
in food processing usually contain materials that derive from the
manufacturing methods used to generate the finished enzyme preparation.
The egg white lysozyme enzyme preparation that is the subject of this
document complies with the general requirements and additional
requirements for enzyme preparations in the Food Chemicals Codex, 4th
ed. (Ref. 14). The egg white lysozyme enzyme preparation that is the
subject of this document may contain substances that are added to the
enzyme preparation, such as preservatives, stabilizers or diluents, and
trace amounts of processing aids that are used in its preparation.
These substances must be acceptable for general use in foods (Refs. 14
and 15).
C. Labeling as a Condition of Use
Egg whites are known to be an allergenic food source, particularly
in children (Ref. 19). There is a literature report (Ref. 20)
indicating that lysozyme may in fact have been an allergen for some
individuals who became sensitive to egg whites. Although the reported
in vitro study does not establish that ingestion of egg white lysozyme
in cheese will actually cause a clinically significant allergic
reaction in such sensitive individuals, FDA is not aware of any data or
information that would refute the study's inference that egg white
lysozyme may be allergenic. Therefore, FDA concludes that there is
insufficient information in the current record to determine whether the
ingestion of egg white lysozyme elicits an allergenic response when
consumed by individuals who are sensitive to egg whites. Accordingly,
as discussed below, FDA is proposing labeling to alert such individuals
to the presence of egg white lysozyme in cheese. Such labeling also
would alert the sensitive population to the possible presence of
source-derived proteins other than lysozyme in the enzyme preparation.
Under section 409(c)(1) of the Federal Food, Drug, and Cosmetic Act
(the act) (21 U.S.C. 348(c)(1)), FDA is authorized, in approving the
use of a food additive, to list the conditions under which the additive
may be safely used. These conditions may include any labeling
requirements that the agency deems necessary to ensure the safe use of
the additive. Similarly, under Sec. 184.1(b)(3) (21 CFR 184.1(b)(3)),
in affirming a substance as GRAS, FDA is authorized to set forth the
particular conditions of use, including labeling, under which there is
general recognition among qualified experts that the use of the
substance is safe. After careful review of the evidence on the use of
egg white lysozyme enzyme preparation in preventing late blowing in
cheese, FDA has tentatively concluded that such use is GRAS only when
the conditions of its use include a declaration on the label or
labeling of the presence of egg white lysozyme in both bulk and
packaged food containing such treated cheese. Therefore, this tentative
final rule (Sec. 184.1550(c)(1)) establishes that the declaration of
egg white lysozyme enzyme preparation by the common or usual name ``egg
white lysozyme'' is a condition of use required for GRAS status, so
that consumers who are allergic to egg white products can be alerted to
the presence of the egg white-derived enzyme in treated cheese.
D. Summary and Conclusions
The petitioner provided published data and information relating to
the identity of, characteristic properties of, and estimated dietary
exposure to the enzyme component (Refs. 1 through 7). The source of the
petitioned enzyme preparation, egg white, has been safely consumed by
humans as a source of food protein throughout recorded history, and,
therefore, is GRAS (Sec. 170.30(d)). The petitioner provided generally
available information showing that the methods used for extracting
lysozyme from the egg white source do not ordinarily alter the chemical
identity and characteristic properties of enzymes (Ref. 8). Moreover,
there is corroborating evidence that the extraction of egg white
lysozyme does not change its chemical identity or characteristics
because the antibacterial activity of egg white lysozyme is retained.
FDA concludes that the methods used to manufacture egg white lysozyme
enzyme preparation do not change the safety for food use of the enzyme
lysozyme and that toxicological
[[Page 12424]]
studies are not necessary to establish the safety of lysozyme or other
source-derived proteins that may remain in the manufactured enzyme
preparation. FDA also concludes that there will be no net increase in
dietary exposure of the general population to the commonly consumed
enzyme lysozyme due to the proposed use in cheese because lysozyme will
simply be transferred from eggs to cheese (Ref. 16).
The petitioner also provided generally available and accepted
information relating to processing aids used in the manufacture of the
enzyme preparation and generally available and accepted specifications
for food grade enzyme preparations (Ref. 14). FDA concludes that
substances added to the egg white lysozyme enzyme preparation or
potential residues of processing aids used in the manufacturing process
do not present a basis for concern about the safety of the egg white
lysozyme enzyme preparation.
The petitioner provided published scientific review articles (Refs.
1 and 2) and a generally available trade bulletin (Ref. 7) that discuss
the use of the egg white lysozyme enzyme preparation in cheese and
other food, including its use for the intended effect of preventing
late blowing of cheese contaminated with C. tyrobutyricum. The
petitioner also provided generally available information documenting
that this intended use of lysozyme has been approved in several
European countries (Refs. 9 through 13). FDA concludes that generally
available and accepted data and information establish that lysozyme
will achieve the intended technical effect of preventing late blowing
in cheese contaminated with C. tyrobutyricum.
Finally, information in the petition and otherwise available to FDA
raises the question of whether the lysozyme component of egg whites is
allergenic. FDA is proposing labeling to alert individuals who may be
sensitive to egg whites to the presence of egg white lysozyme in
cheese, including the possible presence of other source-derived
proteins that may be present in the enzyme preparation.
IV. Comments
FDA received two comments in response to the filing notice. One
comment expressed agreement that lysozyme is GRAS for use in preventing
late blowing in cheese and supported the affirmation of GRAS status by
the agency.
One comment stated that use of lysozyme as a food preservative may
lead to selection of lysozyme-resistant strains of the bacterial food
poisoning agents Listeria monocytogenes and C. botulinum, rendering one
of the body's main defense mechanisms useless against resistant
strains. The comment likened the potential selection of lysozyme-
resistant strains of bacteria to the selection of penicillin-resistant
bacteria as a result of its widespread use. The comment pointed out
that the body could not readily substitute the lysozyme naturally
present in secretions such as tears and saliva for another
antimicrobial.
The mechanism of action of lysozyme involves hydrolysis of the
structural peptidoglycan present in cell walls of susceptible bacteria.
Therefore, development of resistance to lysozyme would require that a
bacterium develop a variant of peptidoglycan that is resistant to the
action of lysozyme. Development of such a variant peptidoglycan is, in
principle, possible. However, as already discussed, lysozyme activity
has been shown to be present in bacteria, fungi, plants, and almost all
animal tissues. If such relative ubiquity has not resulted in the
clinically significant selection of lysozyme-resistant bacteria to
date, the use of lysozyme in those cheeses that are susceptible to late
blowing is unlikely to favor selection of lysozyme-resistant bacteria
and adversely affect the public health. Moreover, FDA is not
considering lysozyme for use as a widespread food preservative. Rather,
FDA is considering the narrow question of whether the use of lysozyme
in preventing late blowing in cheese is generally recognized as safe.
FDA disagrees that this limited use in cheese is analogous to the
widespread use of antibiotics such as penicillin and the subsequent#
selection of antibiotic-resistant bacterial strains. Therefore, FDA
concludes that the use of lysozyme in preventing late blowing in cheese
does not raise concerns about the selection of lysozyme-resistant
strains of L. monocytogenes or C. botulinum.
V. Specifications
The agency finds that, because the potential impurities in the egg
white lysozyme 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 and additional
requirements for enzyme preparations in the monograph on Enzyme
Preparations in the Food Chemicals Codex, 4th ed. (1996), which are
being incorporated by reference in accordance with 5 U.S.C. 552(a) and
1 CFR part 51, are adequate as minimum criteria for food-grade egg
white lysozyme enzyme preparation. Lysozyme assay can be performed
using a method entitled ``Lysozyme hydrochloride, Microbiological
Determination,'' which is included in the petition (Ref. 21) or by
using any appropriate validated method.
VI. Conclusions
The agency has evaluated all available information and finds, based
upon the published information about the manufacturing methods used in
the preparation of egg white lysozyme enzyme preparation, and published
data and information about the identity and characteristic properties
of egg white lysozyme, that the enzyme component of egg white lysozyme
enzyme preparation is unaltered from the lysozyme found in the commonly
consumed food, eggs. The agency also finds, based upon generally
available and accepted information, that when the preparation is
manufactured in accordance with Sec. 184.1550(c), the source, egg
whites, and the manufacturing process will not introduce impurities
into the preparation that may render its use unsafe. Further, the
agency finds, based upon published information, that egg white lysozyme
enzyme preparation will achieve its intended technical effect of
preventing late blowing in cheese contaminated with C. tyrobutyricum.
Therefore, the agency tentatively concludes, based upon the evaluation
of published data and information, corroborated by unpublished data and
information, that the egg white lysozyme enzyme preparation described
in the regulation set out below is GRAS for use by the general
population in preventing late blowing in cheese.
To give interested persons an opportunity to comment on the
proposed label declaration that is a condition of use required for GRAS
status, FDA is issuing this tentative final rule under 21 CFR
10.40(f)(6). FDA will review any comments that are relevant to this
condition of use and that are received within the 75 day comment period
and will respond accordingly to these comments in the Federal Register.
VII. Environmental Considerations
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 finding of
no significant impact and the evidence supporting that finding,
contained in an environmental assessment, may be seen in the Dockets
Management Branch
[[Page 12425]]
(address above) between 9 a.m. and 4 p.m., Monday through Friday.
VIII. Analysis of Economic Impacts
A. Benefit-Cost Analysis
FDA has examined the impacts of this tentative final rule under
Executive Order 12866. Executive Order 12866 directs Federal 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 tentative 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. FDA is tentatively
affirming the GRAS status of egg white lysozyme in cheese only when the
ingredient statement of the bulk and packaged food that contains the
cheese includes the common or usual name of the substance, i.e., ``egg
white lysozyme.'' The labeling requirement will add a small cost to the
future use of the petitioned substance, and therefore, is not a
significant action under the Executive Order 12866.
FDA has examined the impacts of this tentative final rule under the
Unfunded Mandates Reform Act of 1995 (UMRA) (Pub. L. 104-4). A written
statement under section 202(a) of the UMRA is not required for this
rule because the rule does not impose a mandate that results in an
expenditure of $100 million or more by State, local, and tribal
governments in the aggregate, or by the private sector, in any 1 year.
B. Regulatory Flexibility Act
FDA has evaluated this tentative 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.
FDA believes that this tentative final rule is not likely to have a
significant economic impact on a substantial number of small entities.
However, the agency seeks comment on this tentative conclusion. First,
FDA is tentatively affirming the GRAS status of egg white lysozyme in
cheese only when the ingredient statement of the bulk and packaged food
that contains the cheese includes the common or usual name of the
substance, i.e., ``egg white lysozyme.'' This labeling requirement will
impose only minimal costs to the future use of the petitioned
substance. Second, FDA has information that the petitioner does not
currently sell egg white lysozyme in the United States (Refs. 22 and
23). Moreover, FDA is not aware of any manufacture or use of cheese
containing egg white lysozyme in the United States. If no small
entities are currently manufacturing or using cheese containing egg
white lysozyme, the proposed labeling requirements would not impose any
cost to small entities. However, because FDA does not have any
information on whether other entities in the United States are
manufacturing or using cheese containing egg white lysozyme, FDA is
unable to conclude, in this tentative final rule, that there will be no
significant economic impact on a substantial number of small entities.
Therefore, the agency seeks comment on the manufacture or use, by any
small entity, of cheese containing egg white lysozyme. In its final
rule, the agency will, based on any relevant comments received,
determine whether there is a significant economic impact on a
substantial number of small entities.
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. Carini, S., G. Mucchetti, and E. Neviani, ``Lysozyme:
Activity Against Clostridia and Use in Cheese Production--A
Review,'' Microbiologie Aliments Nutrition, 3:299-320, 1985.
2. Procter, V. A., and F. E. Cunningham, ``The Chemistry of
Lysozyme and Its Use as a Food Preservative and a Pharmaceutical,''
CRC Critical Reviews in Food Science and Nutrition, 26:359-395,
1988.
3. Fleming, A., ``On a Remarkable Bacteriolytic Element Found in
Tissues and Secretions,'' Proceedings of the Royal Society, 93:306-
317, 1922.
4. Blake, C. C. F., D. F. Koenig, G. A. Mair, A. C. T. North, D.
C. Phillips, and V. R. Sarma, ``Structure of Hen Egg-White
Lysozyme,'' Nature, 206:757-783, 1965.
5. Kuhara, S., E. Ezaki, T. Fukamizo, and K. Hayashi,
``Estimation of the Free Energy Change of Substrate Binding in
Lysozyme-Catalyzed Reactions,'' Journal of Biochemistry, 92:121-127,
1982.
6. Malcolm, B., K. Wilson, B. Matthews, J. Kirsch, and A.
Wilson, ``Ancestral Lysozymes Reconstructed, Neutrality Tested, and
Thermostability Linked to Hydrocarbon Packing,'' Nature, 345:86-89,
1990.
7. ``The Use of Lysozyme in the Prevention of Late Blowing in
Cheese,'' Bulletin of the International Dairy Foundation, No. 216,
1987.
8. Chaplin, M. F., and C. Bucke, Enzyme Technology, Cambridge
University Press, New York, NY, pp. 66-72, 1990.
9. Denmark Ministry of Agriculture Circular on Approved Enzymes
for Cheese, Journal nr. 10.1.5-18/83, January 13, 1984.
10. French Republic Ministry of Agriculture Service Note 5236,
April 21, 1987.
11. Federal Republic of Germany, Bundesgesetzblatt Nr. 56, pp.
2103-2107, November 22, 1985.
12. Official Gazette of Italian Republic, General Series n. 23,
October 4, 1986.
13. Spanish Ministry of Health and Consumption, General
Direction of Public Health, General Sanitary Register, File n. 8826,
June 20, 1983.
14. Monograph on ``Enzyme Preparations,'' Food Chemicals Codex,
National Academy Press, Washington, DC, 4th ed., pp. 133-134, 1996.
15. Pariza, M. W. and E. M. Foster, ``Determining the Safety of
Enzymes Used in Food Processing,'' Journal of Food Protection,
46:453-468, 1983.
16. Memorandum dated March 20, 1990, from Food and Color
Additives Review Section, FDA, to Direct Additives Branch, FDA,
``Use of Lysozyme to Prevent the `Late Blowing' of Cheese.''
17. Memorandum dated August 5, 1996, from Chemistry Review
Branch, FDA, to Biotechnology Policy Branch, FDA.
18. Kessler, D. A., M. R. Taylor, J. H. Maryanski, E. L. Flamm,
and L. S. Kahl, ``The Safety of Foods Developed by Biotechnology,''
Science, 256:1747-1749 and 1832, 1992.
19. Crespo, J. F., C. Pasqual, A. Ferrer, A. W. Burke, J. M.
Diaz Pena, and M. M. Esteban, ``Egg White-specific IgE Level as a
Tolerance Marker in the Follow-up of Egg Allergy,'' Allergy
Proceedings, 15:73-76, 1994.
20. Anet, J., J. F. Back, R. S. Baker, D. Barnett, R. W. Burley,
and M. E. H. Howden, ``Allergens in the White and Yolk of Hen's Egg;
a Study of IgE Binding by Egg Proteins,'' International Archives of
Allergy and Applied Immunology, 77:364-371, 1985.
21. Lysozyme Hydrochloride, Microbiological Determination.
22. Letter dated November 25, 1996, from John B. Dubeck, Keller
and Heckman, to Linda Kahl, FDA.
23. Letter dated February 28, 1997, from John F. Foley, Keller
and Heckman, to Linda Kahl, FDA.
List of Subjects in 21 CFR Part 184
Food ingredients, Incorporation by reference.
Therefore, under the Federal Food, Drug, and Cosmetic Act and under
[[Page 12426]]
authority delegated to the Commissioner of Food and Drugs and
redelegated to the Director, Center for Food Safety and Applied
Nutrition, it is proposed that 21 CFR part 184 be amended as follows:
PART 184--DIRECT FOOD SUBSTANCES 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 184.1550 is added to subpart B to read as follows:
Sec. 184.1550 Egg white lysozyme.
(a) Egg white lysozyme (CAS Reg. No. 9001-63-2) is the enzyme
peptidoglycan N-acetylmuramoylhydrolase (EC No. 3.2.1.17) obtained by
extraction from egg whites. The enzyme catalyzes the hydrolysis of
peptidoglycan in the cell walls of certain bacteria including
Clostridium tyrobutyricum.
(b) The ingredient meets the general requirements and additional
requirements for enzyme preparations in the monograph on Enzyme
Preparations in the Food Chemicals Codex, 4th ed. (1996), 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., Washington, DC 20418, and 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)(l) The ingredient is used in cheeses, as defined in
Sec. 170.3(n)(5) of this chapter, in accordance with Sec. 184.1(b)(3)
at levels not to exceed current good manufacturing practice.
(2) The affirmation of the use of this ingredient as generally
recognized as safe (GRAS) as a direct human food ingredient is based
upon the following conditions of use:
(i) The ingredient is used as an enzyme as defined in
Sec. 170.3(o)(9) of this chapter.
(ii) Current good manufacturing practice utilizes a level of the
ingredient sufficient to prevent the late blowing of cheeses caused by
the bacterium Clostridium tyrobutyricum during cheese production.
(iii) The ingredient statement for both bulk and packaged food that
contains cheese manufactured using egg white lysozyme shall include the
common or usual name ``egg white lysozyme'' to identify the source of
the protein.
Dated: March 3, 1998.
L. Robert Lake,
Director, Office of Policy, Planning and Strategic Initiatives, Center
for Food Safety and Applied Nutrition.
[FR Doc. 98-6571 Filed 3-12-98; 8:45 am]
BILLING CODE 4160-01-F