Agency Response Letter
GRAS Notice No. GRN 000195

Ms. Lori Gregg
Novozymes North America, Inc.
77 Perry Chapel Church Road
P.O. Box 576
Franklinton, NC 27525

Re: GRAS Notice No. GRN 000195

Dear Ms. Gregg:

The Food and Drug Administration (FDA) is responding to the notice, dated March 1, 2006, 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 this notice on March 3, 2006, filed it on March 3, 2006, and designated it as GRAS Notice No. GRN 000195.

The subject of the notice is mixed β-glucanase and xylanase enzyme preparation from Humicola insolens (H. insolens). The notice informs FDA of the view of Novozymes North America, Inc., (Novozymes) that mixed β-glucanase and xylanase enzyme preparation is GRAS, through scientific procedures, for use in the brewing industry as an enzyme for clarifying beer. The enzyme preparation is intended for use at minimum levels necessary to achieve the desired effect. Novozymes states that the recommended use level of mixed β-glucanase and xylanase preparation (standardized based on β-glucanase activity) is 0.2 grams of enzyme preparation per kilogram of total raw material. The enzyme preparation is added at the mashing stage, and the enzyme activity will be completely inactivated during wort boiling.

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 of the enzyme preparation, and substances used as stabilizers, preservatives or diluents. Enzyme preparations may also contain constituents derived from the production organism and constituents derived from the manufacturing process, e.g., components of the fermentation media or the residues of processing aids. Novozymes' notice provides information about each of the components of the mixed ß-glucanase and xylanase enzyme preparation.

Novozymes provides general information about the identities of ß-glucanase and xylanase. These enzymes are the most important principles in the enzyme preparation and are classified as carbohydrases. The enzyme preparation is used in beer-brewing, primarily during the mashing process to ensure efficient breakdown of ß-glucans, pentosans, and other gums. The enzymes are subsequently heat inactivated and removed by filtration. This reduces the viscosity of the solution, thereby increasing the filtration rate of both wort and beer such that during wort boiling haze is avoided. β-Glucanase (endo-1,4-β -glucanase) is identified by the following classification numbers: EC No. 3.2.1.6 and CAS Registry No. 62213-14-3. Xylanase (endo-1,4-β-xylanase) is identified by the following classification numbers: EC No. 3.2.1.8 and CAS No. 9025-57-4 .

Mixed β-glucanase and xylanase enzyme preparation from H. insolens was reviewed by the Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives (JECFA) at the 61st JECFA (2003) meeting. Also, Canada has approved the use of mixed βß-glucanase and xylanase enzyme preparation produced by H. insolens for use in beer brewing and alcohol production. Mixed ß-glucanase and xylanase enzyme preparation is marketed for use in animal feed in the United States.

The notifier considers the safety of the production organism as the prime consideration in assessing the safety of an enzyme preparation intended for food use. The notice states that a nonpathogenic and nontoxogenic organism can be assumed to produce safe food ingredients when used in conjunction with current good manufacturing practices. Novozymes cites publications as well as a report by the International Food Biotechnology Council as the basis for this conclusion.

The production organism H. insolens is native to the dematiaceous mitosporic fungi (hyphomycetes). The notifier references a published article that states that no reports connecting H. insolens with infectious diseases or mycotoxicosis in humans or animals have been found. The notifier also cites a published article that concludes that H. insolens is nonpathogenic and nontoxogenic.

In its notice, Novozymes cites published comprehensive reviews and studies that support the safety of microbial enzyme preparations used in food processing. Novozymes also presents data from an unpublished 13-week subchronic oral toxicity study in rats and two genotoxicity studies, an in vitro bacterial reverse mutation assay and an in vitro cytogenetic test. Novozymes concludes that these tests show the mixed ß-glucanase and xylanase enzyme preparation does not exhibit mutagenic activity or toxic effect under the specified testing conditions.

Novozymes states that the raw materials used in the fermentation and recovery process for the enzyme concentrate are common ingredients used in the enzyme industry. Novozymes affirms that the quality management system used in the manufacturing process for the enzyme complies with the requirements of International Organization for Standardization 9001. Additionally, Novozymes states that the raw materials are suitable food grade materials. The enzyme preparation is produced following Good Manufacturing Practices (GMPs).

The mixed βglucanase and xylanase enzyme preparation is produced by submerged fed-batch pure culture fermentation of an H. insolens strain. During all steps of fermentation, physical and chemical control measures are taken and microbiological analyses are performed to ensure the absence of foreign microorganisms and to confirm the identity of the production strain. Novozymes states that its specifications for the mixed β-glucanase and xylanase enzyme preparation comply with the purity criteria recommended for enzyme preparations as described in the Food Chemical Codex, 5th Edition, 2003 and conform to the general specifications outlined by JECFA. Novozymes provides specifications for the mixed ß-glucanase and xylanase enzyme preparation including microbiological limits.

Based on the information provided by Novozymes, as well as other information available to FDA, the agency has no questions at this time regarding Novozymes' conclusion that mixed β-glucanase and xylanase enzyme preparation is 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 mixed β-glucanase and xylanase enzyme preparation. As always, it is the continuing responsibility of Novozymes to ensure that food ingredients that the firm ma rkets are safe,and are otherwise in compliance with all applicable legal and regulatory requirements.

In accordance with proposed 21 CFR 170.36(f), a copy of the text of this letter responding to GRN 000195, as well as a copy of the information in this notice that conforms to the information in the proposed GRAS exemption claim (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,

Laura M. Tarantino, Ph.D.
Director
Office of Food Additive Safety
Center for Food Safety and Applied Nutrition