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Collection of Information on Enzymes a Great Deal of Additional Information on the European Union Is Available on the Internet

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Collection of Information on Enzymes a Great Deal of Additional Information on the European Union Is Available on the Internet European Commission Collection of information on enzymes A great deal of additional information on the European Union is available on the Internet. It can be accessed through the Europa server (http://europa.eu.int). Luxembourg: Office for Official Publications of the European Communities, 2002 ISBN 92-894-4218-2 © European Communities, 2002 Reproduction is authorised provided the source is acknowledged. Final Report „Collection of Information on Enzymes“ Contract No B4-3040/2000/278245/MAR/E2 in co-operation between the Federal Environment Agency Austria Spittelauer Lände 5, A-1090 Vienna, http://www.ubavie.gv.at and the Inter-University Research Center for Technology, Work and Culture (IFF/IFZ) Schlögelgasse 2, A-8010 Graz, http://www.ifz.tu-graz.ac.at PROJECT TEAM (VIENNA / GRAZ) Werner Aberer c Maria Hahn a Manfred Klade b Uli Seebacher b Armin Spök (Co-ordinator Graz) b Karoline Wallner a Helmut Witzani (Co-ordinator Vienna) a a Austrian Federal Environmental Agency (UBA), Vienna b Inter-University Research Center for Technology, Work, and Culture - IFF/IFZ, Graz c University of Graz, Department of Dermatology, Division of Environmental Dermatology, Graz Executive Summary 5 EXECUTIVE SUMMARY Technical Aspects of Enzymes (Chapter 3) Application of enzymes (Section 3.2) Enzymes are applied in various areas of application, the most important ones are technical use, manufacturing of food and feedstuff, cosmetics, medicinal products and as tools for re- search and development. Enzymatic processes - usually carried out under mild conditions - are often replacing steps in traditional chemical processes which were carried out under harsh industrial environments (temperature, pressures, pH, chemicals). Technical enzymes are applied in detergents, for pulp and paper applications, in textile manufacturing, leather industry, for fuel production and for the production of pharmaceuticals and chiral substances in the chemical industry. Typically technical enzymes are manufac- tured and used as bulk enzymes in high volumes compared to other areas of enzyme appli- cation. Food enzymes are mainly used in baking industry, for manufacturing fruit juices, in wine making and brewing as well as in cheese manufacturing. An important field of applications in terms of volumes is starch conversion to yield ingredients for foodstuff. The use of enzymes in animal nutrition is an important and growing area of enzyme applica- tion, especially for pig and poultry nutrition. Feed enzymes offer the benefit of degrading specific feed components otherwise harmful or of no value to the livestock. Presently, a number of enzymes are or are intended to be applied in cosmetic products. A well documented use of application is skin peeling, future applications may be skin protection and enzyme systems preserving the cosmetic product. Notable medications of enzymes are as digestive aids, for wound cleaning, lysis of vein thromboses, acute therapy of myocardial infarction and as support in the therapy of certain types of leukaemia. Enzymes can be used in chemical analysis and as a research tool in the life sciences. How- ever, the volumes used in this area of applications are regarded as negligible and this type of enzymes is not considered in the course of this study. Manufacturing of enzymes (Section 3.3) The majority of enzymes currently available is manufactured from microorganism. Manufac- turing process comprise large-scale fermentation to yield high volumes of microbes. En- zymes are either accumulating inside the cells or are secreted into the media of the fermen- tation tanks. In subsequent steps the disrupted cells (or the media including the enzymes) are subjected to further purification processes using variety of chemical, mechanical and thermal techniques (concentration, precipitation, extraction, centrifugation, filtration, chro- matography). The resulting enzyme concentrate is then formulated to the final ready-to-sell product by adding stabilisers, standardizing agents, preservatives and salts. The final en- zyme preparations are usually commercially marketed in granular or liquid forms. The nature of enzyme products (Section 3.4) Within the scope of this study, the terminology for enzymes used by AMFEP was applied. Commercially available enzymes are produced as enzyme concentrates which result from fermentation and subsequent purification steps. The enzyme concentrate contains the active enzyme(s) and various by-products from the fermentation process. The composition and amount of by-products in the enzyme concentrate is extremely variable and depending on the organisms, the media and the conditions applied during fermentation and subsequent 6 Executive Summary downstream processing (30 - 98% by-products in the enzyme concentrate). Thus, in addition to identification and characterisation of the enzyme as the active substance, parameters ap- plicable for characterisation of enzyme concentrate have to be implemented. Additives are added in a subsequent step depending on the particular application and on customers demands (final ready-for-use enzyme preparation). Enzyme preparations should be regarded as preparations in the sense of Directive 1999/45/EC. Recent developments in enzyme manufacturing (Section 3.5) The application of genetic engineering techniques in enzyme manufacturing is dramatically sparking the exploitation of new enzymes and the development of new enzyme properties. Due to new technologies, new enzymes not accessible before can be cloned into and pro- duced from a well-known host organism. Thereby, enzymes from almost any source in na- ture become accessible, including enzymes exhibiting unusual properties, such as extreme thermostability. Applying new technologies, the enzyme properties may be efficiently altered which will lead to an increase in the variability of enzymes available and might lead to en- zymes not present in nature so far. This mainly concerns enzyme stability, catalytic mecha- nism, substrate specificity and range, surface activity, folding mechanisms, cofactor depend- ency, pH- and temperature optima, kinetic parameters. Furthermore, enzymes could be chemically modified. With applying new technologies, the variability in enzyme structure is dramatically increased and enzyme properties are significantly enhanced. Thus, these methods are mainly contrib- uting to technical and economic goals. However, the safety of enzyme manufacturing might also be improved by restricting to few well-known and safe-to-use production strains which are used as hosts for genes from various sources. Industrial Enzymes Presently Marketed in the EU (Chapter 4) A list of all enzymes that are presently marketed in the EU was compiled based on informa- tion given by enzyme-producing industry (AMFEP). This list includes 186 enzymes from 47 different catalytic types and gives information on the catalytic activity, the host and donor organisms and the various fields of application. Regulation of Enzymes in Legislation Depending on their Use (Chapter 5) Enzymes are regulated in different legislation depending on their use. Within the scope of this study, the regulation of enzymes was investigated and analysed regarding legislation for chemicals, food additives and processing aids, animal feed additives, cosmetic products and medicinal products. Chemicals (Section 5.2) The analysis of the national notification systems in the EU, USA, Canada and Australia re- vealed that in these countries industrial enzymes are defined as chemical substances and are consequently subjected to chemicals legislation; information requirements for notifica- tion/reporting are the same as those for chemicals. In all countries, enzymes are described via their catalytic activity. Only in Canada additional information requirements for substances derived from biotechnology (including enzymes) are defined, focussing on the production or- ganism and the enzymatic properties. In the USA, differentiation of enzymes is done on a case-by-case basis depending on the available information, but limited experience has been gained so far. In Australia, assessments on two enzymes were performed and published so far. Executive Summary 7 In the EU, difficulties in enzyme notification were identified along with the differentiation of enzymes, the interpretation of EINECS entries, the decision on notification requirement and the need for the complete testing requirements according to Directive 67/548/EEC. Food additives and processing aids (Section 5.3) In the EU an harmonised authorisation system is only in place for food enzymes used as ad- ditives. Directive 95/2/EEC covers food additives, however, most food enzymes are applied as processing aids, which do not have a technological function in the final foodstuff. Most Member States do not have a national legislation covering these enzymes. International and national committees (SCF, JEFCA, COT), the US FDA and AMFEP issued guidelines on conditions of use, information requirements and safety evaluation of enzymes. These guide- lines pertain both types of enzymes. Information and testing requirements include basic technical data on the enzyme itself, in- formation on the source material, additives and possible contaminants (MO, heavy metals, toxins). Long-term experience in production of a particular enzyme and use in food applica- tion is explicitly taken into account. Safety concerns are mainly focussing on toxic properties of by-products and contaminants. Changes in the production process may lead to a re-evaluation on a case-by-case
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