Agency Response Letter
GRAS Notice No. GRN 000089
Jack Harris
1800 Massachusetts Ave., N.W.
Second Floor
Washington, DC 20036
Re: GRAS Notice No. GRN 000089
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 000089.
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 microorganism (i.e., A. niger), and the method of manufacture.
Carbohydrase enzyme preparation from A. niger is an enzyme preparation obtained from the culture filtrate resulting from a pure culture fermentation of a non-pathogenic and non-toxicogenic strain of A. niger. The preparation contains alpha-amylase (EC 3.2.1.1) or glucoamylase (EC 3.2.1.3). These enzymes catalyze the hydrolysis of starch or starch polysaccharides in food.
Pectinase enzyme preparation from A. niger is an enzyme preparation obtained from the culture filtrate resulting from a pure culture fermentation of a non-pathogenic and non-toxicogenic strain of A. niger. The preparation contains pectin depolymerase (EC 3.2.1.15), pectin methylesterase (EC 3.1.1.11), pectate lyase (EC 4.2.2.2) or pectin lyase (EC 4.2.2.10). These enzymes catalyze the hydrolysis of pectins in fruit-containing products.
Protease enzyme preparation from A. niger is an enzyme preparation obtained from the culture filtrate resulting from a pure culture fermentation of a non-pathogenic and non-toxicogenic strain of A. niger. The preparation contains peptide hydrolases, such as aspartic proteinase (EC 3.4.23.6), which catalyze the hydrolysis of proteins or polypeptides in food.
Glucose oxidase enzyme preparation from A. niger is an enzyme preparation obtained from the intact cells from a pure culture fermentation of a non-pathogenic and non-toxicogenic strain of A. niger. The preparation contains glucose oxidase (EC 1.1.3.4), which catalyzes the removal of glucose or oxygen from food.
Catalase enzyme preparation from A. niger is an enzyme preparation obtained from the intact cells from a pure culture fermentation of a non-pathogenic and non-toxicogenic strain of A. niger. The preparation contains catalase (EC 1.11.1.6), which catalyzes the decomposition of hydrogen peroxide in food.
The source microorganism for each of the enzyme preparations described in GRN 000089 is A. niger. The general taxonomy and characteristics of A. niger are described in standard compendia (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 A. niger. All microbial strains used in enzyme manufacture are started from a pure laboratory culture of A. niger 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 carbohydrase, pectinase, and protease from A. niger are excreted into the fermentation medium. In the submerged culture method, the extracellular location of the enzyme means that no extraction step is needed, and the microorganism and other insoluble matter are removed from the fermentation medium by filtering or centrifuging. In the semisolid culture method, the enzyme is extracted either directly from the moist material, or later after the culture mass has been dried, followed by further processing steps such as clarification, evaporation, precipitation, drying and grinding.
In GRP 3G0016, ETA includes publications that show that glucose oxidase and catalase from A. niger are 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 000089 meets the general and additional requirements in the monograph on enzyme preparations in the Food Chemicals Codex, 4th ed. (1996), pp. 128-135.
The statutory basis for ETA’s determination that carbohydrase, pectinase, protease, glucose oxidase and catalase enzyme preparations from A. niger 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 1952 (Ref. 2), that states that fungal carbohdrase was used in brewing and baking and in the production of corn syrup. ETA also includes an article, published in 1957 (Ref. 3), that states that fungal carbohydrase was used in starch conversion, baking, brewing, syrup production and production of other food products. Although the articles published in 1952 and 1957 do not specify A. niger as the fungal source of carbohydrase, ETA also includes an article published in 1958 (Ref. 4) that states that fungal carbohydrase was used in starch conversion, baking, and processing cereal products, fruit juices and other food products and specifically identifies the source fungus as A. niger.
In GRP 3G0016, ETA includes an article, published in 1952 (Ref. 2), that states that pectic enzyme was used in processing coffee beans. ETA also includes an article, published in 1957 (Ref. 3), that states that fungal pectinase was used in fruit juice and wine processing. Although these articles do not specify A. niger as the fungal source of pectinase, ETA also includes another article, published in 1957 (Ref. 5), that states that fungal pectinase was used in fruit juice processing and coffee bean fermentation and specifically identifies A. niger as the source fungus. ETA also includes an article, published in 1958 (Ref. 4), that states that fungal pectinase was used in processing fruit juices, purees, and coffee beans and identifies the source fungus as A. niger.
In GRP 3G0016, ETA includes an article, published in 1952 (Ref. 2), that states that fungal protease was used in brewing and baking. ETA also includes an article, published in 1957 (Ref. 3), that states that fungal protease was used in baking and beer and ale production. Although the articles published in 1952 and 1957 do not specify A. niger as the fungal source of protease, ETA also includes an article, published in 1958 (Ref. 4), that states that fungal protease was used in baking, processing cereal products, manufacturing beer and ale, and meat tenderizing and specifically identifies the source fungus as A. niger.
In GRP 3G0016, ETA includes an article, published in 1957 (Ref. 3), that states that fungal glucose oxidase was used in de-sugaring of eggs and in processing food items, such as cheese, beer, carbonated beverages, and fruit juices. Although the article published in 1957 does not specify A. niger as the fungal source of glucose oxidase, ETA also includes an article, published in 1958 (Ref. 4), that states that fungal glucose oxidase was used in de-sugaring eggs and manufacturing canned soft drinks and specifically identifies the source fungus as A. niger.
In GRP 3G0016, ETA includes an article, published in 1957 (Ref. 3), that states that fungal catalase was used in food applications (without specifying the food items). Although the article published in 1957 does not specify A. niger as the fungal source of catalase, ETA also includes an article published in 1958 (Ref. 4), that states that fungal catalase was considered for use in cold sterilization of milk for cheese processing and specifically identifies the source fungus as A. niger.
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 carbohydrase, pectinase, protease, glucose oxidase and catalase enzyme preparations from Aspergillus niger 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.
Because the protease enzyme preparation from A. niger would be used to tenderize meat, FDA consulted with the Labeling and Consumer Protection Staff of the Food Safety and Inspection Service of the United States Department of Agriculture (FSIS) during its evaluation of GRN 000089. FSIS has determined that ETA has not provided any data to support the suitability of protease enzyme preparation from A. niger 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. FSIS concludes that ETA needs to provide data that establish that the protease enzyme preparation is being used at the lowest level necessary to achieve the intended technical effect in the specific meat and poultry products to which application is desired. FSIS requests that ETA be advised to seek regulatory guidance from FSIS about the use of protease enzyme preparation from A. niger in meat and poultry products. ETA should direct this 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(1).
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).
| Sincerely, /s/ Alan M. Rulis, Ph.D. Director 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 |
1. Monographs on "Aspergillus niger Group" and "Aspergillus flavus Group", in Raper, K. B. and Fennel, D. I., “The genus Aspergillus,” Williams and Wilkins Company, Baltimore, Maryland, pp. 293-334 and 357-404 (1965).
2. Reed, G., “Industrial enzymes - Now speed natural processes,” Food Engineering, 24: pp. 105-109 (1952).
3. Underkofler, L.A. and W. J. Ferracone, “Commercial enzymes - Potent catalyzers that promote quality,” Food Engineering, 29: pp. 123-133 (1957).
4. Underkofler, L.A., R.R. Barton, and S.S. Rennet, “Microbiological process report - Production of microbial enzymes and their applications,” Applied Microbiology, 6: pp. 212-221 (1958).
5. Kirk, R.E. and Othmer, D.F. (eds.), "Enzymes, Industrial" in Encyclopedia of Chemical Technology, First Supplement Volume, Interstate Cyclopedia, Inc., New York, NY (1957).
(1)FSIS also informed FDA that ETA has not provided any data to support the suitability of carbohydrase, pectinase, glucose oxidase and catalase enzyme preparations from A. niger for use in meat and poultry products. In its notice, ETA does not describe any uses for these enzyme preparations in meat and poultry products.
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