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CFSAN/Office of Food Additive Safety
July 28, 2008
Alice J. Caddow
Genencor, A Danisco Division
925 Page Mill Road
Palo Alto, CA 94304
Re: GRAS Notice No. GRN 000238
Dear Ms. Caddow:
The Food and Drug Administration (FDA) is responding to the notice, dated December 20, 2007, 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 December 26, 2007, filed it on December 27, 2007, and designated it as GRAS Notice No. GRN 000238.
The subject of the notice is a lipase enzyme preparation derived from Hansenula polymorpha expressing a gene encoding a lipase from Fusarium heterosporum (lipase enzyme preparation). The notice informs FDA of the view of Genencor, a Danisco Division (Genencor), that the lipase enzyme preparation is GRAS, through scientific procedures, for use as an enzyme to modify lipids in dough for bakery and noodle applications, in egg yolks for bakery applications and mayonnaise production, and in the refining process of soy bean oil.
Commercial enzyme preparations that are used in food processing typically contain an enzyme component, which catalyzes the chemical reaction that is responsible for its technical effect, as well as 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.
As part of its notice, Genencor includes the report of a panel of individuals (Genencor's GRAS panel) who evaluated the data and information that are the basis for Genencor's GRAS determination. Genencor considers the members of its GRAS panel to be qualified by scientific training and experienced to evaluate the safety of substances added to food. Genencor's GRAS panel evaluated the production organism, the synthetic lipase gene from F. heterosporum, the production process and materials, intended use levels in specified foods, estimates of dietary exposure, as well as unpublished safety studies. Based on the review, Genencor's GRAS panel concluded that lipase enzyme preparation meets its established food grade specifications and is GRAS under the conditions of its intended use.
The notice includes information about identity, activity, and the technical effect of the lipase enzyme. The enzyme is identified by IUB name triacylglycerol lipase, EC No. 18.104.22.168, and Chemical Abstract Service Registry No. 9001-62-1. Genencor notes that the lipase enzyme hydrolyzes ester bonds primarily in the 1- and 3-position of fatty acids in triglycerides. The notifier also states that the lipase enzyme has activity at the sn-1 esters in other lipids, including diacyl-phospholipids and diacyl-galactolipids.
Genencor states that the lipase enzyme preparation conforms to the general and additional requirements for enzyme preparations as described in the Food Chemicals Codex (5th edition) and to the General Specifications for Enzyme Preparations Used in Food Processing as proposed by the FAO/WHO Joint Expert Committee on Food Additives.
Genencor describes the production microorganism of the lipase enzyme, indicating that it is a strain of H. polymorpha known as RB11. Genencor considers the safety of the production organism as the prime consideration in assessing the safety of an enzyme preparation intended for food use. Genencor states that H. polymorpha is a nontoxigenic, nonpathogenic production organism that does not contain pyrogens, pathogens, or viral inclusions. Genencor notes that, because of taxonomic revisions, H. polymorpha is also known as Pichia angusta. Genencor incorporates by reference previous GRAS notices for lipases from Fusarium oxysporum (GRN 000075) and from Thermomyces lanuginosus (GRN 000043), indicating that the activity of its lipase enzyme is identical to these lipases. Genencor further states that its lipase enzyme has approximately 80% amino acid sequence homology with the lipase from F. oxysporum.
Genencor states that its lipase enzyme is manufactured using a submerged, fed-batch, pure culture fermentation of the genetically modified strain of H. polymorpha. The production of each batch of stock culture for fermentation is initiated from a new lyophilized stock culture vial of H. polymorpha production organism. The recovery of the fermented product is described as a multi-step operation consisting of concentration, formulation, and standardization. The final lipase enzyme preparation has specifications for both a dry and liquid enzyme product.
The notifier estimates a maximum daily intake of the lipase enzyme of 0.64 milligrams total organic solids (TOS) per kilogram of body weight per day (mg/kg bw/d) by combining food consumption data from the USDA Continuing Survey of Food Intake by Individuals, 1994-1996 in the United States, and the 1995 Danish Food survey in Denmark with its proposed use levels. In its exposure assessment, the notifier assumes that the lipase enzyme is active in foods, it is not removed or heat-inactivated, and that 100% of foods proposed for use with the lipase enzyme are treated with this enzyme in this estimation.
In its notice, Genencor summarizes unpublished in vivo and in vitro toxicity studies using the lipase enzyme. The notifier concludes that these studies indicate no genotoxic potential and support a no observable adverse effect level of 40 mg total protein/kg bw/d or 669 mg TOS/kg bw/d.
Based on the information provided by Genencor, as well as other information available to FDA, the agency has no questions at this time regarding Genencor's conclusion that the lipase 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 the lipase enzyme preparation. As always, it is the continuing responsibility of Genencor to ensure that food ingredients that the firm markets 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 000238, 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).
Laura M. Tarantino, Ph.D.
Office of Food Additive Safety
Center for Food Safety and Applied Nutrition