[Federal Register Volume 61, Number 41 (Thursday, February 29, 1996)]
[Rules and Regulations]
[Pages 7702-7704]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 96-4629]



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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.

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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.

SUPPLEMENTARY INFORMATION:

I. Background

    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 
notice.

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 
(Sec. 170.30(c)).
    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 
products.
    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 
Sec. 170.30(b).
    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 
preparation.

III. Identity and Technical Effect

    Lactase is the accepted name for the enzyme -D-galactoside 
galactohydrolase (EC 3.2.1.23), 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 
milk products.

[[Page 7703]]


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 
(Ref. 5).
    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).

VI. Conclusions-

    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 
information.
    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 
required.

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 

[[Page 7704]]
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. 
553(d)(1)).

X. 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. 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, 
1981.
    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-
138, 1982.
    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 
22, 1982.
    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 
AS SAFE

    1. The authority citation for 21 CFR part 184 continues to read as 
follows:

    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 
pseudotropicalis.

    (a) This enzyme preparation is derived from the nonpathogenic, 
nontoxicogenic yeast C. pseudotropicalis. It contains the enzyme 
lactase (-D-galactoside galactohydrolase, EC 3.2.1.23), 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 
galactose.
    (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 
such use.

    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