[Federal Register Volume 61, Number 41 (Thursday, February 29, 1996)]
[Rules and Regulations]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 96-4629]
DEPARTMENT OF HEALTH AND HUMAN SERVICES
Food and Drug Administration
21 CFR Part 184
[Docket No. 83G-0062]
Direct Food Substances Affirmed as Generally Recognized as Safe;
Lactase Enzyme Preparation From Candida Pseudotropicalis
AGENCY: Food and Drug Administration, HHS.
ACTION: Final rule.
SUMMARY: The Food and Drug Administration (FDA) is amending its
regulations to affirm that lactase enzyme preparation derived from
Candida pseudotropicalis for use in milk and milk-derived products to
hydrolyze lactose is generally recognized as safe (GRAS). This action
is in response to a petition submitted by Pfizer, Inc.
DATES: Effective February 29, 1996. 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 certain publications listed
in new Sec. 184.1387, effective February 29, 1996.
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.
In accordance with the procedures described in Sec. 170.35 (21 CFR
170.35), Pfizer, Inc., 235 East 42d St., New York, NY 10017, submitted
a petition (GRASP 2G0282) proposing that lactase enzyme preparation
from C. pseudotropicalis be affirmed as GRAS for use as a direct human
food ingredient. (Lactase, the enzyme, is to be distinguished from
lactase enzyme preparation, which contains lactase as the principal
active component but also contains other components derived from the
production organism and fermentation media. This document will refer to
the former as ``lactase'' and to the latter as ``lactase enzyme
preparation.'') Lactase enzyme preparation is used to hydrolyze lactose
in milk and milk products.
FDA published a notice of filing of this petition in the Federal
Register of March 29, 1983 (48 FR 13098), and gave interested persons
an opportunity to submit comments to the Dockets Management Branch
(HFA-305), Food and Drug Administration, 12420 Parklawn Dr., rm. 1-23,
Rockville, MD 20857. FDA received no comments in response to that
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 and ordinarily is to be 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, and
ordinarily is to be based upon generally available data and information
concerning the pre-1958 history of use of the food ingredient
The petition states that C. pseudotropicalis was isolated from
dairy products prior to 1958 (Refs. 1 and 2). Therefore, the petition
argues, lactase produced by the organism has been part of the human
diet for many years and may be presumed to have been in common use in
food prior to January 1, 1958. The petition also states that Pfizer,
Inc., first began commercial production of lactase enzyme preparation
derived from C. pseudotropicalis in 1982 for use in certain dairy
The agency recognizes that C. pseudotropicalis was isolated from
dairy products prior to 1958. However, lactase enzyme preparation
derived from C. pseudotropicalis does not itself have a history of
common use as an ingredient in food before 1958. Therefore, the enzyme
preparation does not qualify for GRAS status based on a history of
common use in food (Sec. 170.30(c)). Accordingly, FDA has evaluated the
enzyme preparation on the basis of scientific procedures under
In evaluating this petition, the agency reviewed information
concerning: (1) The identity and function of the enzyme, (2) the
production and purification of the lactase enzyme preparation, and (3)
the safety of the production organism and the finished lactase enzyme
III. Identity and Technical Effect
Lactase is the accepted name for the enzyme -D-galactoside
galactohydrolase (EC 126.96.36.199), which catalyzes the hydrolysis of the
disaccharide lactose to its component monosaccharides, glucose and
galactose. Lactase enzyme preparations may be produced by fermentation
utilizing any of a large number of microorganisms. A typical example is
the enzyme produced by the yeast Kluyveromyces lactis (Ref. 3).
The lactase preparation that is the subject of this petition is a
soluble enzyme preparation derived from the yeast C. pseudotropicalis
and is composed of the enzyme lactase as the principal active
ingredient, other components derived from the production organism and
the fermentation media, residual amounts of processing aids, and
substances added as stabilizers or diluents. The petitioned enzyme
preparation meets the general and additional requirements for enzyme
preparations found in the Food Chemicals Codex, 3d ed. (1981), which
are incorporated by reference in Sec. 184.1387 (Ref. 4).
Lactase enzyme preparation is intended for use in hydrolyzing
lactose to reduce the lactose content of food products. The petitioner
provided published information to demonstrate that lactase enzyme
preparation from C. pseudotropicalis hydrolyzes lactose in milk and
IV. Production and Purification of Lactase Enzyme Preparation
The lactase enzyme preparation that is the subject of this petition
is produced by controlled aerobic fermentation using a pure culture of
the food-derived yeast C. pseudotropicalis, aseptically grown in a
medium containing suitable food-grade carbohydrates, proteins, mineral
salts, and processing aids. The isolated cells are mixed with a warm
buffer solution consisting of potassium phosphate (mono- and dibasic)
and manganous sulfate and allowed to autolyze for up to 24 hours. The
resulting material is clarified to remove cell debris and other
insoluble solids, and the lactase-containing yeast extract is
concentrated by processes appropriate for food use, including
ultrafiltration. Glycerol and/or sorbitol may be added as stabilizers,
and suitable preservatives may be incorporated during processing. The
stabilized lactase preparation is adjusted to a standard potency using
a combination of water mixed with glycerol or sorbitol.
V. Safety Information-
In its petition, Pfizer, Inc., provided published information to
document that the organism C. pseudotropicalis was isolated from dairy
products as early as 1952 (Refs. 1 and 2). Pfizer, Inc., argues that
since the organism is a copious producer of lactase (Ref. 5), both the
organism and the lactase it produces have been ingested by man for many
years. In addition, Pfizer, Inc., points out that C. pseudotropicalis
resembles K. fragilis (a yeast, also known as Saccharomyces fragilis,
that is approved as a direct food additive ((Sec. 172.896) (21 CFR
172.896))) in all respects except that C. pseudotropicalis is unable to
reproduce sexually (Refs. 1 and 6). K. fragilis, like C.
pseudotropicalis, has been isolated from dairy products (Refs. 1 and
7); in fact, the organisms are often found together in dairy foods
Pfizer, Inc., presented published reports to establish the
similarity between C. pseudotropicalis and K. fragilis. For example, in
an electrophoretic comparison of enzymes, a method used to clarify the
taxonomical and physiological relationships among strains, the
enzymatic patterns of C. pseudotropicalis and its perfect state, K.
fragilis, were shown to coincide (Ref. 8). Further, a study using a
deoxyribonucleic acid (DNA) reassociation technique showed that, within
the accuracy permitted by the technique, C. pseudotropicalis and K.
fragilis have identical DNA sequences (Ref. 9).
The close similarity between the source microorganism (C.
pseudotropicalis) and K. fragilis, which FDA has determined is safe for
use as a direct food additive (Sec. 172.896), supports the safety of
the enzyme preparation (Refs. 10 and 11). Further, the information
submitted by the petitioner establishes that lactase produced by both
yeasts has been ingested by humans for many years with no reported
adverse effects (Ref. 12).
To further document the safety of C. pseudotropicalis, Pfizer,
Inc., presented a published study which compared the pathogenic
potential of several industrial yeasts with that of established
pathogens. The study found that neither C. pseudotropicalis nor K.
fragilis produced signs of tissue invasion or disease. The authors of
the study categorized both organisms in a group of nonpathogenic
organisms (Ref. 13). Finally, Pfizer, Inc., submitted unpublished
corroborative studies conducted in mice to confirm the nonpathogenicity
of C. pseudotropicalis.
After conducting a review of the literature and evaluating these
studies, the agency concludes that C. pseudotropicalis is neither
pathogenic nor toxicogenic (Refs. 14 and 15). Furthermore, the agency
has determined that the autolysis and filtration steps used in
producing and purifying the lactase enzyme preparation effectively
remove viable cells of the production organism (Ref. 15).
Pfizer, Inc., also presented corroborative unpublished toxicity
studies to establish the safety of lactase enzyme preparation derived
from C. pseudotropicalis. These were: (1) An acute oral toxicity study
in rats, (2) mutagenic and cytogenetic assays, and (3) 90-day oral
toxicity studies in rats and dogs. The agency has evaluated the studies
and concludes that the studies showed no evidence of toxicity or
genotoxicity (Ref. 16).
Finally, Pfizer, Inc., presented information regarding use levels
of the enzyme preparation in milk and milk products. Based on this
information, the agency concludes that the use of lactase enzyme
preparation from C. pseudotropicalis would not add to the total
consumption of lactase from all sources because the petitioned enzyme
preparation will be substituted for other lactase enzyme preparations
currently in use (Ref. 17).
The agency has evaluated the information in the petition, along
with other available information, and concludes that lactase enzyme
preparation derived from C. pseudotropicalis is GRAS. This conclusion
is based on published information, corroborated by unpublished data and
Therefore, the agency is affirming that lactase enzyme preparation
derived from C. pseudotropicalis is GRAS with no limits on its
conditions of use other than current good manufacturing practice, in
accordance with 21 CFR 184.1(b)(1). This GRAS affirmation is based on
evaluation of the use of the enzyme preparation to reduce the lactose
content of milk and milk-derived food products.
The agency further finds that because the principal active
ingredient of the enzyme preparation is safe and because expected
impurities in the enzyme preparation do not provide any basis for a
safety concern, the general requirements and additional requirements
for enzyme preparations given in the Food Chemicals Codex, 3d ed.
(1981), pp. 107-110, are adequate as minimum criteria for food-grade
lactase enzyme preparations derived from C. pseudotropicalis.
VII. Environmental Impact
The agency has determined under 21 CFR 25.24(b)(7) that this action
is of a type that does not individually or cumulatively have a
significant effect on the human environment. Therefore, neither an
environmental assessment nor an environmental impact statement is
VIII. Analysis of Impacts
FDA has examined the impact of this final rule under Executive
Order 12866 and the Regulatory Flexibility Act (Pub. L. 96-354).
Executive Order 12866 directs agencies to assess all 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
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 raising novel legal or policy issues. The
Regulatory Flexibility Act requires analyzing options for regulatory
relief for small businesses.
FDA finds that this final rule is not a significant regulatory
action as defined by Executive Order 12866. The final rule does not
raise novel legal or policy
issues. The compliance cost to firms currently in the industry is zero
because the rule prohibits no current activity. Potential benefits
include the wider use of the enzyme preparation because of reduced
uncertainty concerning its regulatory status, and any resources saved
by eliminating the need to prepare further petitions to affirm the GRAS
status of this use of the enzyme preparation.
Finally, in compliance with the Regulatory Flexibility Act, FDA
certifies that the final rule will not have a significant impact on a
substantial number of small businesses. The compliance cost to small
businesses currently in the industry is zero because no current
activity is prohibited under the rule.
IX. 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.
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. Lodder, J., The Yeasts: A Taxonomic Study, 2d ed., pp. 345-
348, 1025-1027, North Holland Publishing Co., Amsterdam, 1970.
2. Rose, A. H., and J. S. Harrison, The Yeasts, V 1, Biology of
Yeasts, p. 139, Academic Press, New York, 1969.
3. 21 CFR 184.1388.
4. ``Monograph on Enzyme Preparations,'' in Food Chemicals
Codex, 3d ed., National Academy Press, Washington, DC, pp. 107-110,
5. Schmidt, J. L., and J. Lenoir, ``Contribution to the Study of
Yeast Flora in Camembert Cheese. Its Development During Ripening,''
Lait 58 (577):355-370, 1978.
6. Kreger van Rij, N. J. W., ``The Yeasts: A Taxonomic Study,''
pp. 233-234, North Holland Pub. Co., Amsterdam, 1970.
7. Lutwick, L. I., H. J. Phaff, and D. A. Stevens,
``Kluyveromyces fragilis as an Opportunistic Fungal Pathogen in
Man,'' Sabouraudia 18:69-73, 1980.
8. Yamazaki, M., and K. Komagata, ``Asporogenous Yeasts and
Their Supposed Ascosporogenous States: An Electrophoretic Comparison
of Enzymes,'' Journal of General and Applied Microbiology, 28:119-
9. Letter from H. J. Phaff, University of California, Davis, to
W. C. Wernau, Pfizer, Inc., February 13, 1980.
10. Memorandum from T. J. McKay, FDA, to M. Custer, FDA,
November 8, 1982.
11. Memorandum from C. B. Johnson, FDA, to N. Beru, FDA,
February 2, 1994.
12. Memorandum from C. B. Johnson, FDA, to the Direct Additives
Branch, FDA, December 6, 1988.
13. Holzschu, D. L. et al., ``Evaluation of industrial yeast for
pathogenicity,'' Sabouraudia 17:71-78, 1979.
14. Memorandum from P. Mislivec, FDA, to M. Custer, FDA, October
15. Memorandum from J. M. Madden, FDA, to M. Peiperl, FDA,
November 5, 1993.
16. Memorandum from A. N. Milbert, FDA, to V. Prunier, FDA,
August 13, 1984.
17. Memorandum from J. Modderman, FDA, to V. Prunier, FDA,
November 7, 1984.
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 SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED
1. The authority citation for 21 CFR part 184 continues to read as
Authority: Secs. 201, 402, 409, 701 of the Federal Food, Drug,
and Cosmetic Act (21 U.S.C. 321, 342, 348, 371).
2. New Sec. 184.1387 is added to subpart B to read as follows:
Sec. 184.1387 Lactase enzyme preparation from Candida
(a) This enzyme preparation is derived from the nonpathogenic,
nontoxicogenic yeast C. pseudotropicalis. It contains the enzyme
lactase (-D-galactoside galactohydrolase, EC 188.8.131.52), which
converts lactose to glucose and galactose. It is prepared from yeast
that has been grown by a pure culture fermentation process.
(b) The ingredient meets the general requirements and additional
requirements for enzyme preparations in the Food Chemicals Codex, 3d
ed. (1981), pp. 107-110, which are 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, 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 limitations other than current good manufacturing
practice. The affirmation of this ingredient as generally recognized as
safe as a direct human food ingredient is based upon the following
current good manufacturing practice conditions of use:
(1) The ingredient is used as an enzyme, as defined in
Sec. 170.3(o)(9) of this chapter, to convert lactose to glucose and
(2) The ingredient is used in food at levels not to exceed current
good manufacturing practice. Current good manufacturing practice is
limited to use of this ingredient to reduce the lactose content in milk
and milk-derived food products where food standards do not preclude
Dated: February 15, 1996.
Fred R. Shank,
Director, Center for Food Safety and Applied Nutrition.
[FR Doc. 96-4629 Filed 2-28-96; 8:45 am]
BILLING CODE 4160-01-F