WORLDWIDE REGULATIONS GOVERNING THE USE OF ENZYMES

VIII. WORLDWIDE REGULATIONS GOVERNING THE USE OF ENZYMES

The authors are more familiar with U.S. laws and regulations regarding enzymes, and views on other countries are not necessarily derived from first-hand knowledge.

A. United States of America

The Food and Drug Administration of the U.S. Department of Health and Human Services has promulgated regulations pertaining to foods, which are published as the Code of Fed- eral Regulations (CFR). This code is divided into 50 titles that represent broad subject areas that come under the scrutiny of federal regulations. Title 21 of CFR pertains to food and drugs. Each title is divided into chapters, which usually bear the name of the issuing agency. Each chapter is subdivided into parts covering specific regulatory areas. Thus most regulations pertaining to foods are 21 CFR 100–199 (Title 21 of the Code of Federal Regulations, Parts 100–199). Part 173 is ‘‘Secondary Direct Food Additives Permitted in Foods for Human Consumption.’’ Subpart B of Part 173 specifically deals with enzyme preparations and microorganisms. Subpart B has nine specific items as follows:

173.110 Amyloglucosidase derived from Rhizopus niveus 173.120 Carbohydrase and cellulase derived from Aspergillus niger 173.130 Carbohydrase derived from Rhizopus oryzae 173.135 Catalase derived from Micrococcus lysodiekticus 173.140 Esterase–lipase derived from Mucor miehei 173.145 α-Galactosidase derived from Morteirella vinaceae var. raffinoseutilizer 173.150 Milk-clotting enzymes, microbial 173.160 Candida guilliermondii 173.165 Candida lypolytica

These enzymes are considered direct food additives. Another group of enzymes are listed in 21 CFR 184, ‘‘Direct Food Substances Af- firmed as Generally Recognized as Safe’’ (GRAS). 21 CFR 184.1027 is mixed carbohy- drase and protease enzyme product; 184.1372 pertains to insoluble glucose isomerase products; 184.1388 addresses lactase enzyme preparation from Kluyveromyces lactis; 184.1585 is papain; 184.1685 is rennet (animal-derived). These lists do not account for some other enzymes that are used in food processing, such as pectinases, which are cov- ered under 21 CFR 101, which deals with food labeling. Subpart F covers exemption from food labeling requirements and specifically under 1.2(ii) makes provisions for processing aids. Processing aids further defined as

1. Substances that are added to a food during the processing of such food but are removed in some manner from the food before it is packaged in its finished

2. Substances that are added to a food during processing that are converted into constituents normally present in the food and do not significantly increases the amount of the constituents naturally found in the food

3. Substances that are added to a food for their technical or functional effect in the finished food at insignificant levels and do not have any technical or func- tional effect in that food

Item 3 is perhaps most fitting for enzymes such as pectinases in the sense that they are used to reduce viscosity and are then destroyed. Tenderization of meat can also be classified in this category or even perhaps under item 2.

Roland (1981) reported that the GRAS Review Branch of the FDA had received

11 GRAS petitions dealing with enzymes of which only one was processed as a food additive. One petition in particular (3-0016) submitted by the Ad Hoc Enzyme Technical Committee covered a large number of microbial animal and plant enzymes, and even after

7 years no action had been taken. A summary of the current status of enzymes in the United States is presented in Table 3 . The Enzyme Technical Association (ETA) is carrying on a dialogue with the FDA in its quest to seek GRAS status for microbial enzymes. Enzymes listed in 21 CFR often specify not only genus and species of microorganisms but also varieties or variants. This position, if adopted by FDA into new regulations, would narrow the definition of accept- able microbes and exclude certain species from the pre-1958 use category. The FDA’s understanding of enzymes would view all microbial enzyme preparations as mixtures of carbohydrases, proteases, and lipases. Further, the status of newer enzymes produced as

a result of genetic manipulation of microorganism is an issue that the FDA is studying. One final note on GRAS status and what it means; the GRAS list reflects FDA opinions as to which substances are generally recognized as safe, but the final authority as to GRAS status does not rest with the FDA but with ‘‘qualified scientists.’’ The Federal Food, Drug and Cosmetic Act does not define the level of scientific training and experience needed to qualify as an expert. Similarly the nature of scientific information used by experts in making a GRAS designation is not specified. Further, the experts need not inform the FDA of the evidence supporting the designation and the agency need not review and approve the evidence before the substances is used in food. Therefore, if the FDA refuses to affirm an enzyme as GRAS, the food industry can still use the enzyme under its own criteria for GRAS. This in no manner prevents the FDA from initiating legal action or enforcement action to stop the use of a non–FDA-approved GRAS enzyme.

B. Canada

Enzymes are treated as food additives and therefore require approval. Approval is obtained by filing a petition in accordance with procedures in Section B.16.002 of the Food and Drug Regulations. The enzymes currently approved (Denner, 1983) are listed in Table 4 . Maximum level of use of enzymes should follow good manufacturing practice. The law also specifies enzyme, source, and permitted uses.

C. European Economic Community

The Association of Microbial Food Enzyme Producers (AMFEP) is the EEC’s equivalent of the U.S. Ad Hoc Enzyme Technical Committee. This organization has formulated gen-

Table 3 Enzymes Permitted for Food Use by the United States Food and Drug Administration Enzyme

Regulatory status α-Amylase

Source

Aspergillus niger

GRAS

A. oryzae

GRAS

Rhizopus oryzae

GRAS

Bacillus stearothermophilus

Petitioned for GRAS

B. subtilis

GRAS

B. licheniformis

GRAS β-Amylase

Barley malt

GRAS Cellulase

Barley malt

Aspergillus niger

GRAS

Petitioned for GRAS α-Galactosidase

Trichoderma reesei

173.145 Glucoamylase or

Morteirella vinaceae var. raffinoseutilizer

GRAS amyloglucosidase

Aspergillus niger

A. oryzae

GRAS

Rhizopus oryzae

173.110 Invertase

R. niveus

GRAS Lactase

Saccharomyces cerevisiae

A. niger

GRAS

A. oryzae

GRAS

Kluyveromyces fragilis

GRAS

Petitioned for GRAS Pectinase

Candida pseudotropicalis

Aspergillus niger

GRAS

173.130 Glucose isomerase

Rhizopus oryzae

184.1372 (immobilized

Streptomyces rubiginosus

184.1372 preparation)

Actinoplanes missouriensis

Streptomyces olivaceus

S. olivochromogenes

Bacillus coagulans

Arthrobacter globiformis

Petitioned for GRAS

Petitioned for GRAS Catalase

Streptomyces murinus

Micrococcus lysodeikticus

A. niger

GRAS

GRAS Glucose oxidase

Bovine liver

GRAS Bromelain

A. niger

GRAS Ficin

pineapples

GRAS Papain

figs

GRAS Glucose oxidase

Papaya

GRAS Bromelain

Aspergillus niger

GRAS Ficin

Pineapples

GRAS Papain

Figs

GRAS Milk-clotting enzyme

Papaya

Endothia parasitica

173.150(a)(1)

Bacillus cereus

173.150(a)(2)

Mucor pusillus Lindt

173.150(a)(3)

173.150(a)(4) Rennet

M. miehei Cooney et Emerson

GRAS Pepsin

Ruminant fourth stomach (abomasum)

GRAS Trypsin

Porcine and bovine stomach

GRAS Esterase/lipase

Porcine, bovine pancrease

Mucor miehei

Bacillus licheniformis

Aspergillus niger

GRAS

GRAS Lipase

Bacillus subtilis

Calf, kid, and lamb pancreatic tissue

GRAS

Aspergillus niger

GRAS

Table 4 Enzymes Permitted for Use in Foods of Canada Enzymes

Permitted uses Bovine rennet

Source

Aqueous extract Cheese, cottage cheese, cream cheese, cream cheese with named additives, cream cheese spread, cream cheese spread with named additives

Bromelain Pineapples Ale, beer, light beer, malt liquor, porter, stout; bread, flour, whole wheat flour, edi- ble collagen; sausage casings, hydrolyzed animal, milk, and vegetable proteins, meat cuts; meat tenderizing, preparations; pumping pickle for curing beef cuts

Ficin Fig tree latex Same uses as bromelain except not used in bread, flour, whole wheat flour, or pump- ing pickles

Papain Fruit of papaya Same as for bromelain and also for beef be- fore slaughter and precooked (instant) ce- reals

Milk-coagulating Mucor miehei , M. pusillus, Cheese, cottage cheese, sour cream, emmen- enzyme

Endothia parasitica taler (Swiss) cheese Pepsin

Glandular layer of porcine Ale, beer, light beer, malt liquor, porter, stomach

stout; cheese, cottage cheese, cream cheese, cream cheese with named ingredi- ents, cream cheese spread, cream cheese spread with named ingredients, defatted soy flour, precooked (instant) cereals

Protease Aspergillus oryzae , A. niger, Same as for bromelain and also dairy-based Bacillus subtilis

flouring preparations, distillers mash, in- dustrially spray-dried cheese powder, pre- cooked (instant) cereals, unstandardized bakery foods

Rennet Aqueous extracts from fourth Same as for bovine rennet and in unstan- stomach of calves, kids, or

dardized milk-based dessert preparations lambs Lipase

Aspergillus niger , A. oryzae; Dairy-based flavoring preparations, liquid edible forestomach tissue

and dried egg white, Romano cheese of calves, kids, or lamb; animal pancreatic tissue

Lipoxidase Soybean whey or meal

Bread, flour

Pancreatin Pancrease of the hog or ox Liquid and dried egg white, precooked (in- stant) cereals, starch used in the produc- tion of dextrins, maltose, dextrose, or glu- cose, or glucose solids

Catalase Aspergillus niger , Micrococ- Soft drinks, egg albumin cus lysodeikticus bovine liver

Glucose oxidase Aspergillus niger Soft drinks, liquid whole egg, egg white, and liquid egg yolk destined for drying Cellulase

Aspergillus niger Distillers mash, liquid coffee concentrate, spice extracts, natural flavor and color ex-

Table 4 Continued Enzymes

Permitted uses Hemicellulase

Source

Bacillus subtilis Ale, beer, light beer, malt liquor, porter, corn for degerming distillers mash des- tined for vinegar manufacture, unstandard- ized bakery products

Pentosanase Aspergillus niger Ale, beer, light beer, malt liquor, porter, stout, corn for degerming distillers mash, mash destined for vinegar manufacture, unstandardized bakery products

Pectinase Aspergillus niger , Rhizopus Cider; wine; distillers mash; juice of named oryzae

fruits; natural flavor and color extrac- tives; skin of citrus fruits destined for jam, marmalade, and candied fruit produc- tion; vegetable stock for use in soup ma- nufacture

Glucanase Aspergillus niger , Bacillus Same as for pentosanase subtilis Invertase

Saccharomyces Soft-centered and liquid-centered confec- tions, unstandardized baking foods Lactase

Aspergillus niger , Rhizopus Lactose-reducing enzyme preparations, milk oryzae , Saccharomyces

destined for use in ice cream mix Glucoamylase

Aspergillus niger , A. oryzae, Ale, beer, light beer, malt liquor, porter, (amyloglucosi-

stout, bread, flour, whole wheat flour, dase maltase)

chocolate syrups, distillers mash, pre- cooked (instant) cereals, starch used in the production of dextrins, maltose, dex- trose, glucose (glucose syrups), glucose solids, unstandardized bakery products

Distillers mash Glucose solids, unstandardized bakery prod-

ucts

Rhizopus niveus

Distillers mash

Rhizopus delemar Mash destined for vinegar making Multiplici sporus

Brewers mash, distillers mash, mash for vin- egar making, starch used in production of dextrins, maltose, dextrose, glucose (glu- cose syrups), glucose solids (dried glu- cose syrups)

Another advisory trade group is the Association of Manufactures of Animal/Plant- Derived Food Enzymes (AMAFE), which unlike AMFEP concerns itself with only en- zymes from nonmicrobial sources. Like the Food Chemical Codex in the United States the EEC organization also relies on data submitted and compiled by the FAO/WHO Joint Expert Committee on Food Additives (JECFA, 1981). The JECFA data reflect both FCC and AMAFE/AMFEP advice on enzymes. Individual EEC member states have different

1. Belgium This country considers enzymes as processing aids, and authorization is needed for their

use. The list of permitted enzymes is narrow; an attempt was made to enlarge and revise this list by royal decree, but this attempt was unsuccessful.

2. Denmark This largest producer of enzymes in the world requires that 6 months prior to the use of

any enzyme a notification to and registration with the Danish National Food Institute be completed. The director of the Institute can approve the use within a 6-month period, but if at the end of this period the director has not responded to the notification, then approval is automatically assumed. The procedures for notification and registration of enzyme prod- ucts were first published in 1974 and are being revised.

3. Federal Republic of Germany Enzymes are considered to be food additives. No authorization and sanctions are required,

except for cheese, fruit juices, and wine.

4. France Decrees are necessary for using food additives such as enzymes. In addition memos are

issued periodically declaring the ‘‘toleration’’ of specific enzymes not sanctioned by prior decree. Applications for the use of enzymes must be filed with Service de la Repression des Frandes.

5. Greece, Ireland, and the Netherlands These countries have no specific controls on the use of enzymes.

6. Italy Enzymes are considered to be processing aids, and their use in beer and wine is controlled

by decree.

7. United Kingdom Enzymes are deemed to be processing aids, and as such there are no controls. The Food

Additive and Contaminants Committee has recommended that controls be instituted for the use of enzymes. A prior sanction for certain enzymes and sources was also recom- mended ( Table 5 ).

The list in Table 5 does not consider lipooxygenase, catalase from Micrococcus lysodeikticus , hemicellulases, ficin, β-amylase, pentosanase, or esterases as recommended for food use. Additionally, certain microorganisms such as Penicillium funiculosum, P. lilacinum , P. emersonii, Klebsiella aerogenes, and Streptomyces fradiae, which are not recognized as safe organisms in the United States or Canada, would be permitted for use in the United Kingdom.

8. Japan Additives can be classified as natural or synthetic. If a naturally occurring compound is

modified or synthesized, then it becomes a synthetic additive. For example, citric acid

Table 5 Food Additives and Contaminants Committee Recommended Prior Sanctioned Enzymes and Sources of Such Enzymes in the United Kingdom

Enzymes Sources Acid proteinase (including pepsin

Porcine gastric mucosa; abomasum of calf or and chymosin)

lamb; adult bovine abomasum; Mucor miehei, M. pusillus

α-Amylase Porcine or bovine pancreatic tissue, Aspergillus ni- ger , A. oryzae, Bacillus licheniformis, B. subtilis Bromelain

Ananas bracteatus , A. comosus Catalase

Bovine liver, Aspergillus niger Cellulase

Aspergillus niger , Trichoderma viride Dextranase

Penicillium funiculosum , P. lilacinum Endothia carboxyl proteinase

Endothia parasitica

Fructofuranosidase (invertase) Saccharomyces cerevisiae Galactofuranosidase (invertase)

Aspergillus niger

Endo-1,3(4)- β-D-glucosidase (lami- Aspergillus niger , Bacillus subtilis, Penicillum em- narinase)

ersonii

Glucose isomerase

Bacillus coagulans

Glucose isomerase (immobilized) Bacillus coagulans , Streptomyces olivaceous Glucose oxidase

Aspergillus niger

Exo-1,4-D-glucosidase (gluco-

Aspergillus niger

amylase) Exo-1, 4-D-glucosidase (immobi-

Aspergillus niger

lized) Neutral proteinase

Aspergillus oryzae , Bacillus subtilis Papain/chymopapain

Carica papaya

Pectin esterase

Aspergillus niger

Pectin lyase

Aspergillus niger

Polygalacturonase

Aspergillus niger

Pullulanase

Klebsiella aerogenes

Serine proteinase (including ltrypsin) Porcine or bovine pancratic tissue, Bacillus licheni- formis , Streptomyces fradiae Triacylglycerol lipase

Edible oral forestomach tissues of the calf, kid, or lamb; porcine or bovine pancreatic tissue

neutralization is the same as chemically synthesized citric acid and therefore is considered synthetic. If an additive falls into the natural category, it needs no approval and listing.

The Food sanitation Investigation Council in accordance with food sanitation laws is involved in the approval process. In order for a compound to be approved the applicant must show that

1. The additive is demonstrably safe.

2. The additive is advantageous to consumer in that it satisfies one or more of the following conditions: (a) it is necessary in the manufacturing process, (b) it helps to maintain the nutritional value of the food, (c) it prevents or minimizes degradation of food, and (e) it reduces the price to the consumer.

3. The additive can be shown to be either superior to or more useful than existing

4. The additive can be chemically identified by analysis in the food as sold to the consumer.

If the additive (1) could disguise poor quality in either the basic food or manufacturing process, (2) impairs nutritional quality, (3) acts as curative or therapeutic, or (4) causes an improvement that could be duplicated by modifying the manufacturing process, it will

be rejected. Nutritional aspects of food additives receive more explicit attention, and there is a list of specific permitted nutritional additives. Once an additive passes preliminary scru- tiny, the next step is to determine toxicological safety at proposed levels of use. Such testing must be done at two authentic research institutes in Japan or at such institute if reliable supporting evidence is available from foreign literature sources.