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CFSAN/Office of Food Additive Safety
April 3, 2002
Enzyme Technical Association
1800 Massachusetts Ave., N.W.
Washington, DC 20036
Re: GRAS Notice No. GRN 000088
Dear Mr. Harris:
The Food and Drug Administration (FDA) is responding to the notice, dated November 8, 2001, that you submitted in accordance with the agency’s proposed regulation, proposed 21 CFR 170.36 (62 FR 18938; April 17, 1997; Substances Generally Recognized as Safe (GRAS); the GRAS proposal). FDA received the notice on November 16, 2001, and designated it as GRAS Notice Nos. GRN 000088, GRN 000089 and GRN 000090 (as explained below).
The subject of the notice is a group of ten microbially derived enzyme preparations, i.e., carbohydrase, pectinase, protease, glucose oxidase and catalase enzyme preparations from Aspergillus niger, carbohydrase and protease enzyme preparations from Aspergillus oryzae, carbohydrase enzyme preparation from Rhizopus oryzae, invertase enzyme preparation from Saccharomyces cerevisiae, and lactase enzyme preparation from Kluyveromyces marxianus. The notice informs FDA of the view of the Enzyme Technical Association (ETA) that these enzyme preparations are GRAS, through common use in food, for use as enzymes in catalyzing specific reactions in the processing of food. Each of the enzyme preparations is used at levels not to exceed current good manufacturing practice. These enzyme preparations are also the subjects of a GRAS affirmation petition (GRP 3G0016) submitted by the Ad Hoc Enzyme Technical Committee (now known as ETA) to FDA in 1973 and amended a few times thereafter. In its notice, the ETA requested that FDA convert the filed GRAS affirmation petition GRP 3G0016 for these ten enzyme preparations to a GRAS notice in accordance with the agency’s proposed regulation, proposed 21 CFR 170.36 (62 FR 18938; April 17, 1997; Substances Generally Recognized as Safe (GRAS))
For administrative expediency, FDA divided ETA’s GRAS notice into three separate GRAS notices. GRN 000088 includes invertase from Saccharomyces cerevisiae, and lactase from Kluyveromyces marxianus. GRN 000089 includes carbohydrase, pectinase, protease, glucose oxidase and catalase from Aspergillus niger. GRN 000090 includes carbohydrase and protease from Aspergillus oryzae, and carbohydrase from Rhizopus oryzae. In this letter, FDA responds to GRN 000088.
Commercial enzyme preparations that are used in food processing typically contain an enzyme component, which catalyzes the chemical reaction that is responsible for the technical effect, as well as substances used as stabilizers, preservatives or diluents. Enzyme preparations may also contain constituents derived from the source organism and constituents derived from the manufacturing process, e.g., components of the fermentation media or the residues of processing aids. In GRP 3G0016, ETA includes data and information about the technical effect of each enzyme component, the source microorganisms (i.e., S. cerevisiae and K. marxianus), and the method of manufacture.
Identity and Technical Effect
Invertase enzyme preparation from S. cerevisiae is an enzyme preparation obtained from the intact cells from a pure culture fermentation of a non-pathogenic and non-toxicogenic strain of S. cerevisiae. The preparation contains the enzyme invertase (EC 220.127.116.11), which catalyzes the hydrolysis of sucrose to glucose and fructose.
Lactase enzyme preparation from K. marxianus is an enzyme preparation obtained from the intact cells from a pure culture fermentation of a non-pathogenic and non-toxicogenic strain of K. marxianus. The preparation contains the enzyme lactase (EC 18.104.22.168), which catalyzes the hydrolysis of lactose to glucose and galactose.
Source Microorganism and Method of Manufacture
The source microorganisms for the invertase enzyme preparation and the lactase enzyme preparation described in GRN 000088 are S. cerevisiae and K. marxianus, respectively. The general taxonomy and characteristics of S. cerevisiae are described in standard compendia (Refs. 1 and 2). The general taxonomy and characteristics of K. marxianus (formerly known as Saccharomyces fragilis and Kluyveromyces fragilis) are described in standard compendia (Refs. 1 and 2). A monograph well accepted for its standards of nomenclature has reclassified the yeast S. fragilis into K. marxianus (Ref. 1).
In GRP 3G0016, ETA includes publications that describe generally accepted microbiological techniques that are used in the manufacture of the enzyme preparations from S. cerevisiae and K. marxianus. All microbial strains used in enzyme manufacture are started from a pure laboratory culture and grown in a sterile liquid nutrient medium or sterile moistened semisolid medium. Generally accepted microbiological techniques are used to exclude contaminating organisms and to avoid development of substrains from within the culture itself. Although specific conditions of fermentation vary from manufacturer to manufacturer, common fermentation procedures, which have been described in the literature, are: (1) the submerged culture method, and (2) the semisolid culture method. During fermentation by either method, the pH, temperature, disappearance of certain ingredients, purity of culture, and level of enzyme activity are carefully controlled. The fermentation is harvested at the point where laboratory tests indicate that maximum production of enzyme activity has been attained.
In GRP 3G0016, ETA includes publications that show that each of the enzymes described in GRN 000088 is retained in the cells in the fermentation medium. In the submerged culture method, the cells are removed by filtration before further processing. In both the submerged culture method and the semisolid culture method, the cells are autolyzed in water containing calcium chloride and/or phosphates. The cell debris is removed by filtration or centrifugation, yielding a clear solution for further processing. Further processing steps may involve clarification, evaporation, precipitation, drying and grinding.
Each of the enzyme preparations described in GRN 000088 meets the general and additional requirements in the monograph on enzyme preparations in the Food Chemicals Codex, 4th ed. (1996), pp. 128-135.
Evidence of Common Use in Food Before 1958
The statutory basis for ETA’s determination that invertase from S. cerevisiae and lactase from K. marxianus are GRAS for their intended use is through experience based on common use in food before 1958. Under 21 CFR 170.30(c)(1), general recognition of safety through experience based on common use in food is based solely on food use of the substance prior to January 1, 1958, and ordinarily is based upon generally available data and information.
In GRP 3G0016, ETA includes an article, published in 1957 (Ref. 3), that states that yeast invertase was used in producing artificial honey and invert sugar. Although the article published in 1957 does not specify S. cerevisiae as the source of invertase, ETA also includes another article, published in 1957 (Ref. 4), that states that yeast invertase was used in manufacturing confectionary products and specifically identifies S. cerevisiae as the source yeast. ETA also includes an article, published in 1958 (Ref. 5), that states that yeast invertase was used in manufacturing artificial honey and invert sugar and identifies the source yeast as S. cerevisiae.
In GRP 3G0016, ETA includes an article, published in 1952 (Ref. 6), that states that lactase was used in the production of ice cream. ETA also includes an article, published in 1957 (Ref. 3), that states that yeast lactase was suggested for use in the production of ice cream to prevent lactose crystallization. Although these articles do not specify K. marxianus as the source of yeast lactase, ETA also includes another article, published in 1957 (Ref. 4), that states that yeast lactase was used in production of milk products and specifically identifies S. fragilis (reclassified as K. marxianus) as the source yeast. ETA also includes an article, published in 1958 (Ref. 5), that states that yeast lactase was used in the manufacture of ice cream and identifies the source yeast as S. fragilis.
Based on the information provided by ETA, as well as the information in GRP 3G0016 and other information available to FDA, the agency has no questions at this time regarding ETA’s conclusion that invertase enzyme preparation from S. cerevisiae and lactase enzyme preparation from K. marxianus are GRAS under the intended conditions of use. The agency has not, however, made its own determination regarding the GRAS status of the subject use of these enzyme preparations. As always, it is the continuing responsibility of each manufacturer to ensure that food ingredients that the firm markets are safe, and are otherwise in compliance with all applicable legal and regulatory requirements.
Consultation with the Food Safety and Inspection Service of the U. S. Department of Agriculture
Because the intended use of invertase enzyme preparation from S. cerevisiae and lactase enzyme preparation from K. marxianus is for general use in food, FDA consulted with the Labeling and Consumer Protection Staff of the Food Safety and Inspection Service of the United States Department of Agriculture (FSIS) about potential applications of these enzyme preparations in meat or poultry products. FSIS has determined that ETA has not provided any data to support the suitability of invertase enzyme preparation from S. cerevisiae and lactase enzyme preparation from K. marxianus for use in meat and poultry products. Suitability relates to the effectiveness of an ingredient in performing the intended purpose of use and the assurance that the conditions of use will not result in an adulterated product or one that misleads consumers. If either of these enzyme preparations has application in meat or poultry products, ETA would need to provide data that establish that the ingredient is being used at the lowest level necessary to achieve the intended technical effects in the specific meat and poultry products to which application is desired. ETA would direct its inquiry to Dr. Robert Post, Director, Labeling and Consumer Protection Staff, Office of Policy, Program Development and Evaluation, Food Safety and Inspection Service, 300 12th Street, SW, Room 602, Washington, DC 20250-3700. The telephone number of his office is (202) 205-0279 and the telefax number is (202)205-3625.
In accordance with the interim policy discussed in the GRAS proposal (62 FR 18938 at 18954), FDA has not committed any resources to review of GRP 3G0016 since November 16, 2001, the date that we received your conversion request.
In accordance with proposed 21 CFR 170.36(f), a copy of the text of this letter, as well as a copy of the information in ETA’s notice that conforms to the information in proposed 21 CFR 170.36(c)(1), is available for public review and copying on the homepage of the Office of Food Additive Safety (on the Internet at http://www.cfsan.fda.gov/~lrd/foodadd.html).
Alan M. Rulis, Ph.D.
Office of Food Additive Safety
Center for Food Safety and Applied Nutrition
cc: Dr. Robert Post, Director
Labeling and Consumer Protection Staff
Office of Policy, Program Development and Evaluation
Food Safety and Inspection Service
300 12th Street, SW, Room 602
Washington, DC 20250-3700
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