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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Link to relevant study record(s)

Description of key information

No bioaccumulation potential.

Key value for chemical safety assessment

Bioaccumulation potential:
no bioaccumulation potential

Additional information

In general toxicokinetic studies performed on enzymes are very limited, but toxicokinetic information can be derived from the structure of enzymes combined with knowledge available for proteins in general since enzymes are proteins with catalytic activity.

Skin absorption

The physic-chemical properties of a compound are decisive for the potential percutaneous penetration, in particular factors like ionization, molecular size and lipophilicity. In general, non-ionized molecules easily penetrate the skin, with small molecules penetrating more easily than large molecules. Lipophilicity also facilitates penetration. Investigations of percutaneous absorption of peptides, proteins and other molecules of large size revealed that percutaneous absorption of proteins is extremely low and of no toxicological relevance [1 -2].

Gastrointestinal tract absorption

Proteins are digested into amino acids by pancreatic proteolytic enzymes in the lumen of the gastrointestinal tract [3]. As enzymes are simply a class of proteins, it can be expected that enzymes will undergo the same process, simply representing a tiny food source. Furthermore, enzymes have been used for decades in treatment of both adults and children with exocrine pancreatic insufficiency. Typical enzymatic drugs (e.g. Creon® from Solvay Pharmaceuticals or Pancrease Microtabs from Jansson/Cilaq) are a combination of the enzymes amylase, lipase and protease – enzymes, which are also used in a wide range of industrial applications. These drugs are typically administered orally at therapeutic concentrations i.e. concentrations where a digestive effect can be expected. Clinical trials and crossover studies confirmed the safe use of these compounds in patients, both adults and children, confirming the low toxicity of enzymes [4-15]. Additionally, in a study investigating the gastrointestinal absorption of enzymes in pigs with pancreatic insufficiency and treated with Creon®, analysis of plasma samples taken in the period of 0.5 to 48 hours after oral administration of the drug, did not result in treatment related changes of plasma enzyme levels indicating no gastric absorption of the administered enzymes [16]. Furthermore, a variety of enzymes are added to animal feed with the purpose to increase nutrient digestibility in the gastrointestinal tract. The safety of such enzymes active in the gastrointestinal tract are thoroughly evaluated as part of their approval process in the EU and elsewhere.

Lysozyme is approved as food additive (beer[18], wine[19 -20] and Cheese[21]) and will be transferred to the Community list of food enzymes as soon as this list is drawn up.

In conclusion, from the available data combined with the knowledge of the fate of proteins in the gastrointestinal system, it can be concluded that absorption of enzymes in toxicological significant amounts through the gastrointestinal tract is unlikely.

Respiratory tract absorption

Enzymes can be inhaled in the form of small dust particles or aerosols i.e. adhered to solid dust particles or as droplets of fluid. Absorption of hydrophilic substances such as enzymes by lung tissue is determined by diffusion and depends on molecular size. The transport channels in the alveolar membrane have a size of 1 nm (10Å) [3], which excludes the absorption of enzymes, since their size is above 1 nm. Removal of deposits depends on the site of deposition. In the alveoli where the main removal is via phagocytosis [3], the macrophages carrying the deposits can move to the interstitium, the ciliated epithelium or the lymphatic system indicating that there could be a risk of systemic exposure to enzymes by this route. However, due to the fact that enzymes are potential respiratory allergens, stringent risk management strategies have been introduced for the working environment leading to very low pulmonary exposure excluding any chance of toxicologically significant absorption. Consumer exposure is even lower. Furthermore, no bioaccumulation will occur after absorption due to rapid biological degradation and enzymes hydrophilic nature. In conclusion, absorption of enzymes by the respiratory tract can be considered insignificant.

Bioavailability

Due to the combined information that skin absorption of enzymes is at a toxicologically insignificant level, that enzymes are degraded in the gastrointestinal tract and that they are only absorbed to a very low extent by the respiratory tract, the total bioavailability of enzymes can be concluded to be extremely low. This is further supported by the physic-chemical data, i.e. enzymes have a low logKOW value, indicating that they have no bioaccumulation potential and can be anticipated to be readily biodegraded. Thus, systemic exposure following enzyme exposure at occupational and consumer exposure levels is without toxicological significance.

Lysozyme was first discovered by A. Fleming, who identified lysozyme as an antibacterial enzyme present in nasal mucus membrane.

Luce and Pellegrini (1976 [17]) have reported in an handbook the conclusions reached by Fleming. About the metabolism they reported that Lysozyme can cross the intestinal barrier without structural modification, since the molecular fragments would not have reacted with the antilysozyme antibodies.

The pharmacokinetics of lysozyme after administration by oral route in rats is that the exogenous enzyme was in appreciable quantities in the portal blood. The absorption by lymphatic route is negligible.

Furthermore it was reported that Lysozyme concentrates particularly in the liver and kidneys, the principal organs of its metabolization and elimination.

Lysozyme is eliminated by the renal route. The lowest quantity recovered in the urine compared to the administration dose might be explained with the storage of the substance in some organs or to metabolizazion into enzymatically inactive amino acid chains which are therefore not microbiologically detectable.

Reference

[1] Basketter,D.A., English,J.S., Wakelin,S.H., and White,I.R. (2008) Enzymes, detergents and skin: facts and fantasies. British journal of dermatology 158, 1177-1181

[2] Pease,C.K.S., White,I.R., and Basketter,D.A. (2002) Skin as a route of exposure to protein allergens. Clinical and experimental dermatology 27, 296-300

[3] Niesink,R.J.M., deVries,J., and Hollinger,M.A. (1996) Toxicology, Principles and Applications CRC Press, Inc. and Open University of The Netherlands.

[4] Barra,E., Stolarczyk,A., Socha,J., Oralewska,B., Kowalska,M., Skoczen,M., and Wawer,Z. (1998) Efficacy of enzyme supplementation in children with cystic fibrosis. Pediatria Polska 73, 177-182

[5] Borowitz,D., Goss,C.H., Stevens,C., Hayes,D., Newman,L., O'Rourke,A., Konstan,M.W., Wagener,J., Moss,R., Hendeles,L., Orenstein,D., Ahrens,R., Oermann,C.M., Aitken,M.L., Mahl,T.C., Young,K.R., Dunitz,J., and Murray,F.T. (2006) Safety and preliminary clinical activity of a novel pancreatic enzyme preparation in pancreatic insufficient cystic fibrosis patients. Pancreas 32, 258-263

[6] Borowitz,D., Goss,C.H., Limauro,S., Konstan,M.W., Blake,K., Casey,S., Quittner,A.L., and Murray,F.T. (2006) Study of a novel pancreatic enzyme replacement therapy in pancreatic insufficient subjects with cystic fibrosis. Journal of Pediatrics 149, 658-662

[7] Dominguez,M., Iglesias-Garcia,J., Iglesias-Rey,M., Figueiras,A., and Vilarino-Insua,M. (2005) Effect of the administration schedule on the therapeutic efficacy

[8] Halm, U., Löser, C., Löhr, M., Katschinski, M., and Mössner, J. (1999) A double-blind, randomized, multicentre, crossover study to prove equivalence of pancreatin minimicrospheres versus microspheres in exocrine pancreatic insufficiency. Alimentary pharmacology & therapeutics 13, 951-957

[9] Heubi, J.E., Boas, S.R., Blake, K., Nasr, A.R.H., Woo, M.S., Graff, G.R., Hardy, K.A., maro-Galvez, R., Latino, M., and Lee, C. (2008) Zentase, a novel pancreatic enzyme product (Pep), is effective in mild, moderate, and severe exocrine pancreatic insufficiency (Epi). Gastroenterology 134, A583-A584

[10] Keller, J. and Layer, P. (2006) Are monolithic enteric-coated enzyme preparations effective in pancreatic exocrine insufficiency? A multicentre, double blind, placebo controlled cross-over trial. Gastroenterology 130, A517

[11] Konstan, M.W., Stern, R.C., Trout, J.R., Sherman, J.M., Eigen, H., Wagener, J.S., Duggan, C., Wohl, M.E.B., and Colin, P. (2004) Ultrase MT12 and ultrase MT20 in the treatment of exocrine pancreatic insufficiency in cystic fibrosis: Safety and efficacy. Alimentary Pharmacology and Therapeutics 20, 1365-1371

[12] Konstan, M.W., Liou, T.G., Strausbaugh, S., Ahrens, R.C., Kanga, J.F., Graff, G.R., Moffett, K.S., Millard, S., Nasr, S.Z., Vezina, M., Spenard, J., and Grondin, J. (2008) Efficacy and safety of Ultrase (R) MT20 in treating pancreatic insufficiency in cystic fibrosis. Gastroenterology 134, A228-A229

[13] Laake, K. (1980) ENZYMIC DRUGS. Side Effects of Drugs Annual 222-225

[14] Patchell, C.J., Desai, M., Weller, P.H., Macdonald, A., Smyth, R.L., Bush, A., Gilbody, J.S., and Duff, S.A. (2002) Creon 10,000 Minimicrospheres vs. Creon 8,000 microspheres--an open randomised crossover preference study. Journal of cystic fibrosis: official journal of the European Cystic Fibrosis Society 1, 287-291

[15] Saeed, Z., Wojewodka, G., Marion, D., Guilbault, C., and Radzioch, D. (2007) Novel pharmaceutical approaches for treating patients with cystic fibrosis. Current Pharmaceutical Design 13, 3252-3263

[16] Gewert,K., Holowachuk,S.A., Rippe,C., Gregory,P.C., Erlanson-Albertsson,C., Olivecrona,G., Kruszewska,D., Piedra,J.V., Weström,B., and Pierzynowski,S.G. (2004) The enzyme levels in blood are not affected by oral administration of a pancreatic enzyme preparation (Creon 10,000) in pancreas-insufficient pigs. Pancreas 28, 80-88

[17] Luce Barbara, Pellegrini Rinaldo, “Fleming lysozyme. Biological Significance and therapeutic applications”. Ed. Minerva Medica. 1976.

[18] Commision regulation (EU) No 471/2012 of 4 June 2012, amending Annex II to Regulation (EC) No 1333/2008 of the European Parliament and of the Council as regards the use of lysozyme (E 1105) in beer.

[19] Commision regulation (EU) No 2066/2001 of 22 October 2001; amending Regulation (EC) No 1622/2000 as regards the use of lysozyme in wine products.

[20] Commision regulation (EU) No 606/2009 of 10 July 2009, laying down certain detailed rules for implementing Council Regulation (EC) No 479/2008 as regards the categories of grapevine products, oenological practices and the applicable restrictions.

[21] European Parliament and council directive No 95/2/EC of 20 February 1995, on food additives other than colours and sweeteners.