Safety evaluation of the food enzyme lysozyme from hens' eggs

Abstract

The food enzyme lysozyme (peptidoglycan N‐acetylmuramoylhydrolase; EC 3.2.1.17) is produced with hens' eggs and submitted by DSM Food Specialties BV. It is intended to be used in brewing processes, milk processing for cheese production as well as wine and vinegar production. The dietary exposure to the food enzyme–total organic solids (TOS) was estimated to be up to 4.9 mg TOS/kg body weight per day. This exposure is lower than the intake of the corresponding fraction from eggs, for all population groups. Egg lysozyme is a known food allergen. The Panel considered that, under the intended conditions of use, the residual amounts of lysozyme in treated beers, cheese and cheese products as well as wine and wine vinegar may trigger adverse allergenic reactions in susceptible individuals. Based on the data provided, the origin of the food enzyme and an exposure to the food enzyme comparable to the intake from eggs, the Panel concluded that the food enzyme lysozyme does not give rise to safety concerns under the intended conditions of use, except for the known adverse allergic reactions that occur in susceptible individuals.

1. Introduction

Article 3 of the Regulation (EC) No 1332/20081 provides definition for ‘food enzyme’ and ‘food enzyme preparation’.

‘Food enzyme’ means a product obtained from plants, animals or microorganisms or products thereof including a product obtained by a fermentation process using microorganisms: (i) containing one or more enzymes capable of catalysing a specific biochemical reaction; and (ii) added to food for a technological purpose at any stage of the manufacturing, processing, preparation, treatment, packaging, transport or storage of foods.

‘Food enzyme preparation’ means a formulation consisting of one or more food enzymes in which substances such as food additives and/or other food ingredients are incorporated to facilitate their storage, sale, standardisation, dilution or dissolution.

Before January 2009, food enzymes other than those used as food additives were not regulated or were regulated as processing aids under the legislation of the Member States. On 20 January 2009, Regulation (EC) No 1332/2008 on food enzymes came into force. This Regulation applies to enzymes that are added to food to perform a technological function in the manufacture, processing, preparation, treatment, packaging, transport or storage of such food, including enzymes used as processing aids. Regulation (EC) No 1331/20082 established the European Union (EU) procedures for the safety assessment and the authorisation procedure of food additives, food enzymes and food flavourings. The use of a food enzyme shall be authorised only if it is demonstrated that:

• it does not pose a safety concern to the health of the consumer at the level of use proposed;

• there is a reasonable technological need;

• its use does not mislead the consumer.

All food enzymes currently on the European Union market and intended to remain on that market, as well as all new food enzymes, shall be subjected to a safety evaluation by the European Food Safety Authority (EFSA) and approval via an EU Community list.

The ‘Guidance on submission of a dossier on food enzymes for safety evaluation’ (EFSA, 2009a) lays down the administrative, technical and toxicological data required.

1.1. Background and terms of reference as provided by the requestor

1.1.1. Background as provided by the European Commission

Only food enzymes included in the Union list may be placed on the market as such and used in foods, in accordance with the specifications and conditions of use provided for in Article 7 (2) of Regulation (EC) No 1332/2008 on food enzymes.

A joint application has been introduced by the Association of Manufactures and Formulators of Enzyme Products (AMFEP) for the re‐evaluation, modification of the specifications and extension of use of the authorisation of the food enzyme lysozyme from hen's egg.

Following the requirements of Article 12.1 of Commission Regulation (EU) No 234/2011 implementing Regulation (EC) No 1331/2008, the Commission has verified that the application falls within the scope of the food enzyme Regulation and contains the relevant elements required under Chapter II of that Regulation.

1.1.2. Terms of reference

The European Commission requests the European Food Safety Authority to carry out the safety assessment on the food enzyme lysozyme from hen's egg in accordance with Article 17.3 of Regulation (EC) No 1332/2008 on food enzymes.

The application was submitted initially as a joint dossier3 and identified as the EFSA‐Q‐2015‐00395. During the risk assessment phase, it was found that the technical dossier is too generic to be evaluated. A solution was found on 16 March 2020 via an ad‐hoc meeting between EFSA, the European Commission and representatives from the Association of Manufacturers and Formulators of Enzyme Products (AMFEP).4 It was agreed that joint dossiers will be split into individual data packages.

The current opinion addresses one data package originating from the joint dossier EFSA‐Q‐2015‐00395. This data package, identified as EFSA‐Q‐2022‐00009, concerns the food enzyme lysozyme that is produced from hen's egg and submitted by DSM Food Specialties.

2. Data and methodologies

2.1. Data

The applicant has submitted a dossier in support of the application for authorisation of the food enzyme lysozyme from hen's egg. The dossier was submitted on 22 December 2021.

Additional information was requested from the applicant during the assessment process on 14 March 2022 and was consequently provided 22 April 2022 (see ‘Documentation provided to EFSA’).

2.2. Methodologies

The assessment was conducted in line with the principles described in the EFSA ‘Guidance on transparency in the scientific aspects of risk assessment’ (EFSA, 2009b) and following the relevant guidance documents of the EFSA Scientific Committee.

The current ‘Guidance on the submission of a dossier on food enzymes for safety evaluation’ (EFSA, 2009a) has been followed for the evaluation of the application with the exception of the exposure assessment, which was carried out in accordance to the updated ‘Scientific Guidance for the submission of dossiers on food enzymes’ (EFSA CEP Panel, 2021a).

3. Assessment

IUBMB nomenclature	Lysozyme
Systematic name	Peptidoglycan N‐acetylmuramoylhydrolase
Synonyms	Muramidase, globulin G, mucopeptide glucohydrolase
IUBMB No	EC 3.2.1.17
CAS No	12650‐88‐3
EINECS No	235‐747‐3

Lysozymes catalyse the hydrolysis of (1–4)‐β‐linkages between N‐acetylmuramic acid and N‐acetyl‐D‐glucosamine residues in peptidoglycans of bacterial cell walls. The food enzyme under application is intended to be used in brewing processes, milk processing for cheese production as well as for wine and wine vinegar production.

3.1. Source of the food enzyme

The lysozyme is extracted from the eggs of hens (Gallus gallus domesticus).5 These eggs fulfil all requirements of European legislation covering the trade of eggs and egg products intended for human consumption.

3.2. Production of the food enzyme

The food enzyme is manufactured according to the Food Hygiene Regulation (EC) No 852/20046, with food safety procedures based on Hazard Analysis and Critical Control Points, and in accordance with current good manufacturing practice.7

On receipt from suppliers, eggs are ■■■■■ broken to separate the yolk from the egg white. The egg white is passed through ■■■■■ column designed to selectively adsorb lysozyme. The column is then washed ■■■■■ to remove the albumin fraction. The lysozyme is then eluted from the column ■■■■■, precipitated by pH adjustment and recovered by filtration. The recovered enzyme is mixed ■■■■■, further purified, concentrated as lysozyme hydrochloride and pasteurised. Finally, the lysozyme hydrochloride may be spray‐dried for use in a granular form or diluted for use in a liquid form.8 The applicant provided full information on the identity of all substances used in the manufacturing process of the food enzyme.9 An amount of ■■■■■ liquid egg white is needed to produce ■■■■■ of lysozyme (on dry weight),10 corresponding to a yield factor of ■■■■■.

The Panel considered that sufficient information has been provided on the manufacturing process and the quality assurance system implemented by the applicant to exclude issues of concern.

3.3. Characteristics of the food enzyme

3.3.1. Properties of the food enzyme

Egg white lysozyme consists of a single polypeptide chain of 129 amino acids with a calculated mass of 14.4 kDa (Canfield, 1963; Cagielska‐Radziejewska et al., 2008).11

The in‐house determination of lysozyme activity is based on the reduction in optical density of a Micrococcus luteus (formally M. lysodeikticus) suspension. The activity is expressed in FIP units. One FIP Unit is defined as the amount of lysozyme that causes a decrease in absorbance of 0.001 per minute at 450 nm, 25°C and pH 7.0.12

The food enzyme has a temperature optimum around 50°C (pH 6.5) and is active across a broad pH range (pH 4–9), with an optimum at 6.5. Thermostability was tested after a pre‐incubation for 30 min at different temperatures up to 100°C. The activity was reduced by 80% after pre‐incubation at 50°C and was no longer detected after pre‐incubation at 70°C (Nawaz et al., 2022).

3.3.2. Chemical parameters

Data on the chemical parameters of the food enzyme were provided for six batches (three in granular and three in liquid form).13 The average enzyme activity of the three batches of the granular form was 41,200 FIP U/mg and that of the three batches of the liquid form 9,700 FIP U/mg. The activity/TOS ratio of the three batches of the granular form was 43,187 FIP U/mg TOS and that of the three batches of the liquid form 45,458 FIP U/mg TOS.

The applicant produces the food enzyme in a way to meet the specifications for lysozyme authorised as a food additive (E 1105) under Regulation (EU) No 231/2012. The specifications require a minimum lysozyme hydrochloride content of 950 mg/g on a dry matter basis. The analytical data provided demonstrated compliance with the specifications (Table 1).

Table 1.

Composition of the food enzyme

Parameters	Unit	Batches (granular form)			Batches (liquid form)
		1	2	3	4	5	6
Lysozyme activity	FIP U/mg (a)	41,900	41,000	40,700	9,700	10,100	9,300
Protein	%	88.9	87.6	90.0	14.3	20.4	20.7
Ash	%	1.1	0.2	0.2	2.2	0.1	0.1
Water	%	3.5	4.7	4.1	78.0	77.7	77.7
Total organic solids (TOS) (b)	%	95.4	95.1	95.7	19.8	22.2	22.2
Lysozyme activity/TOS	FIP U/mg TOS	43,920	43,113	42,528	48,989	45,495	41,891

^(a)FIP Unit: Fédération Internationale Pharmaceutique Unit (see Section 3.3.1).

^(b)TOS calculated as 100% – % water – % ash.

3.3.3. Purity

The lead content in the six batches was below 5 mg/kg,14 which complies with the specification for lead as laid down in the general specifications for enzymes used in food processing (FAO/WHO, 2006). In addition, the levels of arsenic and mercury were below the limits of detection (LoDs) of the employed methodologies. ^{14 ,} 15

The food enzyme complies with the microbiological criteria for total coliforms, Escherichia coli and Salmonella, as laid down in the general specifications for enzymes used in food processing (FAO/WHO, 2006). ¹⁴

The Panel considered that the information provided on the purity of the food enzyme is sufficient.

3.4. Toxicological data

According to the Commission Implementing Regulation (EU) No 562/201216, an application for the safety evaluation of a food enzyme does not need to include toxicological data if the food enzyme is obtained from edible parts of an animal intended or reasonably expected to be ingested by humans.

According to the EFSA ‘Scientific Guidance for the submission of dossiers on Food Enzymes’, for food enzymes derived from plants and animals that are consumed by the European population, two criteria must be met: (i) No hazard is introduced through the manufacturing process, and (ii) when it can be demonstrated that the dietary exposure to the food enzyme–TOS is within the same magnitude as the dietary intake of the fraction of the plant or animal material comparable to the food enzyme–TOS (EFSA CEP Panel, 2021a).

The Panel considered that these requirements are fulfilled, because:

1. the safety of lysozyme has already been evaluated by JECFA (1992) and it is authorised as a food additive in the EU;

2. the manufacturing process of the food enzyme does not introduce substances that could raise safety concerns;

3. the compositional and purity data provided were considered sufficient;

4. eggs, the source of lysozyme, are extensively consumed throughout the EU and elsewhere in the world;

5. the dietary exposure to the food enzyme–TOS under the intended conditions of use was within the same magnitude compared to the intake from eggs (see Section 3.5).

3.4.1. Allergenicity17

The allergenicity assessment considered only the food enzyme and not any carrier or other excipient that may be used in the final formulation.

Egg allergy is one of the most frequent allergies encountered in the European population. Lysozyme as a component of the egg white is recognised to contribute to the allergic response in sensitised individuals (Bianchi, 1982; Hoffman, 1983; JECFA, 1992). In 2011, EFSA concluded that wines treated with lysozyme may trigger adverse allergic reactions in susceptible individuals (EFSA, 2011).

Two human studies with egg‐allergic individuals undergoing skin prick testing with lysozyme‐treated wines demonstrated significant residual amounts of lysozyme in treated wines, but provided no information about the clinical reactivity of egg‐allergic individuals to wines treated with lysozyme when consumed (Kirschner et al., 2009; Weber et al., 2009).

Other studies, including a Double Blind Placebo‐Controlled Food Challenge study upon oral challenge with lysozyme in allergenic children (Fremont et al., 1997), demonstrated allergic reactions to lysozyme‐treated cheese (Malmheden Yman, 2004), but these results are inconclusive with respect to the likelihood of clinical allergic reactions to oral consumption of lysozyme in egg‐allergic subjects due to the inadequate clinical characterisation of the study population.

The Panel concluded that, under the intended conditions of use, the residual amounts of lysozyme in treated beers, cheese and cheese products as well as wine and wine vinegar may trigger adverse allergenic reactions in susceptible individuals.

3.5. Dietary exposure

3.5.1. Intended use of the food enzyme

The food enzyme is intended to be used in three food processes at the recommended use levels summarised in Table 2.18

Table 2.

Intended uses and recommended use levels of the food enzyme as provided by the applicant ¹⁸

Food manufacturing process (a)	Raw material (RM)	Recommended dosage of the food enzyme (mg TOS/kg RM) (b)
Brewing processes	Cereals	140–290
Milk processing for cheese production	Milk	15–45
Wine and wine vinegar production (c)	Grapes	50–480

^(a)The description has been harmonised by EFSA according to the ‘EC working document describing the food processes in which food enzymes are intended to be used’ – not yet published at the time of adoption of this opinion.

^(b)The numbers in bold were used for calculations.

^(c)The applicant confirmed the use of this lysozyme in the production of wine vinegars. ¹⁰

Lysozyme is predominately effective against Gram‐positive bacteria, including lactic acid bacteria. In the brewing process, lactic acid bacteria and some clostridia cause off‐flavours. The lysozyme can be added at various stages at which undesirable bacteria have the opportunity to grow. Consequently, lysozyme may be used to treat the raw material, such as barley, hops, the brewer's yeast before addition to the fermenter or the wort prior to or during the fermentation.19

Lysozyme is used in cheese production predominately to prevent a phenomenon referred to as ‘late blowing’. This is caused mainly by the growth of Clostridium tyrobutyricum, a contaminant of the milk used for cheese production. C. tyrobutyricum ferments the lactate present in cheese, resulting the production of CO₂. The accumulation of such gases during the later stages of curing, particularly in pressed cheeses, may cause the cheese to ‘blow’.20

In wine making, lysozyme is used to control the growth of Gram‐positive spoilage bacteria, to control aspects of malolactic fermentations and to stabilise wines after alcoholic fermentation or when malolactic fermentation is completed. Its use potentially reduces the need for high levels of sulfur dioxide and may help to minimise the concentration of biogenic amines in the final product.21

Based on the temperature profile of the food enzyme (see Section 3.3.1), the food enzyme may remain active in cheese, beer and wine, depending on the processing conditions.

3.5.2. Dietary exposure estimation

Following the EFSA Guidance Document on food enzymes (EFSA CEP Panel, 2021a), a comparison was made between the chronic exposures:

• dietary exposure to the food enzyme–TOS resulting from the intended use by the applicant (herein referred as ‘FE‐TOS’); and

• dietary exposure to a fraction in hen's egg comparable to the food enzyme–TOS resulting from the consumption of eggs or egg‐derived foods (herein referred to as source material TOS equivalent, ‘SMT–Equivalent’).

Chronic exposure to the food enzyme–TOS was calculated by combining the maximum recommended use level with individual consumption data (EFSA CEP Panel, 2021a). The estimation involved selection of relevant food categories and application of technical conversion factors (EFSA CEP Panel, 2021b). Exposure from all FoodEx categories was subsequently summed up, averaged over the total survey period (days) and normalised for body weight. This was done for all individuals across all surveys, resulting in distributions of individual average exposure. Based on these distributions, the mean and 95th percentile exposures were calculated per survey for the total population and per age class. Surveys with only 1 day per subject were excluded and high‐level exposure/intake was calculated for only those population groups in which the sample size was sufficiently large to allow calculation of the 95th percentile (EFSA, 2011).

Table 3 provides an overview of the derived exposure estimates across all surveys. Detailed mean and 95th percentile exposure to the food enzyme–TOS per age class, country and survey, as well as contribution from each FoodEx category to the total dietary exposure are reported in Appendix A – Tables 1 and 2. For the present assessment, food consumption data were available from 48 dietary surveys (covering infants, toddlers, children, adolescents, adults and the elderly), carried out in 26 European countries (Appendix B). The highest dietary exposure was estimated to be about 4.9 mg TOS/kg body weight (bw) per day in the elderly people at the 95th percentile.

Table 3.

Summary of estimated dietary exposure to food enzyme–TOS in six population groups

Population group	Estimated exposure (mg TOS/kg body weight per day)
	Infants	Toddlers	Children	Adolescents	Adults	The elderly
Age range	3–11 months	12–35 months	3–9 years	10–17 years	18–64 years	≥ 65 years
Min–max mean (number of surveys)	0.045–0.284 (12)	0.132–0.510 (15)	0.115–0.257 (19)	0.063–0.170 (21)	0.222–1.039 (22)	0.142–1.462 (23)
Min–max 95th percentile (number of surveys)	0.142–0.971 (11)	0.282–1.492 (14)	0.241–0.706 (19)	0.135–0.580 (20)	0.940–3.632 (22)	0.674–4.900 (22)

The chronic dietary exposure to the SMT–Equivalent was calculated by first estimating the intake of hen's egg from all dietary sources (applying recipe and conversion fractions reported in Appendix C). The intake of egg was converted to egg white, using a factor of 60% to account for the amount of egg white in an egg.22 This intake was further converted into a fraction comparable to the food enzyme–TOS via application of a yield factor (■■■■■) provided by the applicant to take into account the yield of the food enzyme‐TOS from egg white (Section 3.2).

Table 4 provides an overview of the estimated exposure to the SMT‐Equivalent. Mean and 95th percentile exposure to the SMT‐Equivalent per age class, country and survey are reported in Appendix A – Table 3. The contribution of the SMT‐Equivalent from each FoodEx category to the total dietary exposure is indicated in Appendix A – Table 4. The highest dietary exposure was estimated to be about 17.609 mg TOS/kg bw per day in toddlers at the 95th percentile.

Table 4.

Summary of estimated dietary exposure to the SMT‐Equivalent, resulting from the consumption of egg white in six population groups

Population group	Estimated exposure (mg/kg body weight per day)
	Infants	Toddlers	Children	Adolescents	Adults	The elderly
Age range	3–11 months	12–35 months	3–9 years	10–17 years	18–64 years	≥ 65 years
Min–max mean (number of surveys)	0.059–1.484 (12)	0.805–5.404 (15)	1.080–3.547 (19)	0.580–2.109 (21)	0.452–1.238 (22)	0.300–1.535 (23)
Min–max 95th percentile (number of surveys)	0–8.913 (11)	4.228–17.609 (14)	2.921–11.133 (19)	2.136–6.829 (20)	1.795–4.868 (22)	1.645–3.240 (22)

The intake of the FE‐SMT (Table 4) is comparable to the dietary exposure to the food enzyme‐TOS (Table 3) for all the population groups.

3.5.3. Uncertainty analysis

In accordance with the guidance provided in the EFSA opinion related to uncertainties in dietary exposure assessment (EFSA, 2006), the following sources of uncertainties have been considered and are summarised in Table 5.

Table 5.

Qualitative evaluation of the influence of uncertainties on the dietary exposure estimate

Sources of uncertainty	Direction of impact
	Exposure to FE–TOS	Exposure to SMT–Equivalent
Model input data
Consumption data: different methodologies/representativeness/underreporting/misreporting/no portion size standard	+/−	+/−
Use of data from food consumption survey of a few days to estimate long‐term (chronic) exposure for high percentiles (95th percentile)	+	+
Possible national differences in categorisation and classification of food	+/−	+/−
Model assumptions and factors
Exposure to FE–TOS was calculated based on the maximum recommended use level	+	NA
Selection of broad FoodEx categories for the exposure assessment	+	+
Despite the exclusion of wine vinegar as an intended use, EFSA included not only wine, but also wine vinegar in the calculation.	+	NA
Use of recipe fractions in disaggregation FoodEx categories	+/−	+/−
Use of technical factors in the exposure model	+/−	+/−
Selection of FoodEx categories likely to contain egg for the intake assessment of SMT‐equivalent, based on the national food descriptors	NA	+/−
The applied yield factor was the mean value	NA	+/−

NA: not applicable.

+: uncertainty with potential to cause overestimation of exposure.

–: uncertainty with potential to cause underestimation of exposure.

Both estimates are derived using the same food consumption data and exposure model and, hence, share a number of uncertainties, which do not have an effect on the comparison of the two estimates.

3.6. Margin of exposure

Since no toxicological assessment was considered necessary by the Panel, the margin of exposure was not calculated.

4. Conclusion

Based on the data provided, the origin of the food enzyme and an exposure to the food enzyme comparable to the intake from eggs, the Panel concluded that the food enzyme lysozyme does not give rise to safety concerns under the intended conditions of use, except for the known adverse allergic reactions that occur in susceptible individuals.

5. Documentation as provided to EFSA

Application for authorization of lysozyme from hens' egg. December 2021. Submitted by DSM Food Specialties BV.

Additional information. April 2022. Submitted by DSM Food Specialties BV.

Abbreviations

bw

body weight

CAS

Chemical Abstracts Service

CEF

EFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing Aids

CEP

EFSA Panel on Food Contact Materials, Enzymes and Processing Aids

EINECS

European Inventory of Existing Commercial Chemical Substances

FAO

Food and Agricultural Organization of the United Nations

IUBMB

International Union of Biochemistry and Molecular Biology

JECFA

Joint FAO/WHO Expert Committee on Food Additives

kDa

kiloDalton

LoD

limit of detection

TOS

total organic solids

WHO

World Health Organization

Appendix A – Dietary exposure estimates to the food enzyme–TOS and the SMT–Equivalent in details

Information provided in this appendix is shown in an Excel file (downloadable https://efsa.onlinelibrary.wiley.com/doi/10.2903/j.efsa.2023.7916#support‐information‐section).

The file contains four sheets, corresponding to four tables:

Table 1: Mean and 95th percentile exposure to the FE‐TOS per age class, country and survey.

Table 2: Contribution of FoodEx categories to the FE‐TOS dietary exposure.

Table 3: Mean and 95th percentile exposure to the SMT‐Equivalent per age class, country and survey.

Table 4: Contribution of FoodEx categories to the SMT‐Equivalent dietary exposure.

Appendix B – Population groups considered for the exposure assessment

Population	Age range	Countries with food consumption surveys covering more than 1 day
Infants	From 12 weeks on up to and including 11 months of age	Bulgaria, Cyprus, Denmark, Estonia, Finland, France, Germany, Italy, Latvia, Portugal, Slovenia, Spain
Toddlers	From 12 months up to and including 35 months of age	Belgium, Bulgaria, Cyprus, Denmark, Estonia, Finland, France, Germany, Hungary, Italy, Latvia, Netherlands, Portugal, Republic of North Macedonia*, Serbia*, Slovenia, Spain
Children	From 36 months up to and including 9 years of age	Austria, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Italy, Latvia, Netherlands, Portugal, Republic of North Macedonia, Serbia*, Spain, Sweden
Adolescents	From 10 years up to and including 17 years of age	Austria, Belgium, Bosnia and Herzegovina*, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Italy, Latvia, Montenegro*, Netherlands, Portugal, Romania, Serbia*, Slovenia, Spain, Sweden
Adults	From 18 years up to and including 64 years of age	Austria, Belgium, Bosnia and Herzegovina*, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Ireland, Italy, Latvia, Montenegro*, Netherlands, Portugal, Romania, Serbia*, Slovenia, Spain, Sweden
The elderly (a)	From 65 years of age and older	Austria, Belgium, Cyprus, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Ireland, Italy, Latvia, Montenegro*, Netherlands, Portugal, Romania, Serbia*, Slovenia, Spain, Sweden

^(a)The terms ‘children’ and ‘the elderly’ correspond, respectively, to ‘other children’ and the merge of ‘elderly’ and ‘very elderly’ in the Guidance of EFSA on the ‘Use of the EFSA Comprehensive European Food Consumption Database in Exposure Assessment’ (EFSA, 2011).

^*Consumption data from these pre‐accession countries are not reported in Table 3 of this opinion; however, they are included in Appendix B for testing purpose.

Appendix C – FoodEx categories used to derive intake estimates of SMT‐Equivalent and the respective conversion factors

FoodEx code	FoodEx name	Hierarchical level	Fraction of egg in FoodEx category (a)	Proportion of egg white in egg (b)	Percentage of FoodEx containing egg
A.01.05.003	Noodle, wheat flour, with eggs	3	0.14	0.6	1
A.01.05.005	Pasta, wheat flour, with eggs	3	0.2	0.6	1
A.01.05.007	Pasta, wheat flour, filled	3	0.12	0.6	0.5
A.01.05.014	Pasta, wheat wholemeal, with eggs	3	0.19	0.6	1
A.01.07	Fine bakery wares (unspecified)	4	0.1	0.6	1
A.01.07.001	Pastries and cakes (unspecified)	4	0.1	0.6	1
A.01.07.001.001	Beignets	4	0.051	0.6	1
A.01.07.001.002	Buns	4	0.1	0.6	0.5
A.01.07.001.003	Cake from batter	4	0.261	0.6	1
A.01.07.001.004	Cheese cream cake	4	0.1769	0.6	1
A.01.07.001.005	Cheese cream sponge cake	4	0.3176	0.6	1
A.01.07.001.006	Chocolate cake	4	0.0987	0.6	1
A.01.07.001.007	Chocolate cake with fruits	4	0.05922	0.6	1
A.01.07.001.008	Cream cake	4	0.151	0.6	1
A.01.07.001.009	Cream cheese cake	4	0.13	0.6	1
A.01.07.001.010	Cream custard cake	4	0.01305	0.6	1
A.01.07.001.011	Cream custard sponge cake	4	0.01305	0.6	1
A.01.07.001.012	Croissant	4	0.1319	0.6	1
A.01.07.001.013	Croissant, filled with chocolate	4	0.098925	0.6	1
A.01.07.001.014	Croissant, filled with cream	4	0.100244	0.6	1
A.01.07.001.015	Croissant, filled with jam	4	0.098925	0.6	1
A.01.07.001.016	Croquembouche	4	0.120492	0.6	1
A.01.07.001.017	Doughnuts	4	0.0271	0.6	1
A.01.07.001.018	Éclair	4	0.177391	0.6	1
A.01.07.001.019	Flan	4	0.18	0.6	1
A.01.07.001.020	Fruit cake	4	0.152685	0.6	1
A.01.07.001.022	Cheese pie	4	0.04	0.6	1
A.01.07.001.024	Gingerbread	4	0.066	0.6	1
A.01.07.001.025	Gougere	4	0.251025	0.6	1
A.01.07.001.026	Kringles	4	0.042	0.6	1
A.01.07.001.027	Nut cream cake	4	0.233621	0.6	1
A.01.07.001.028	Pancakes	4	0.12	0.6	1
A.01.07.001.029	Profiterole	4	0.123839	0.6	1
A.01.07.001.030	Pyramid cake	4	0.1831	0.6	1
A.01.07.001.032	Scone	4	0.063	0.6	1
A.01.07.001.033	Sponge dough	4	0.261	0.6	1
A.01.07.001.034	Sponge cake	4	0.261	0.6	1
A.01.07.001.035	Sponge cake roll	4	0.17487	0.6	1
A.01.07.001.036	Muffins	4	0.2667	0.6	1
A.01.07.001.037	Waffles	4	0.1442	0.6	1
A.01.07.001.039	Cream‐cheese strudel	4	0.012	0.6	1
A.01.07.001.040	Cheese pastry goods from puff pastry	4	0.1319	0.6	1
A.01.07.001.041	Croissant from puff pastry	4	0.1319	0.6	1
A.01.07.001.042	Brioche	4	0.14	0.6	1
A.01.07.001.043	Macaroons	4	0.8	1	1
A.01.07.001.044	Lebkuchen	4	0.066	0.6	1
A.01.07.001.045	Dumpling	4	0.076	0.6	1
A.01.07.001.046	Cake marbled, with chocolate	4	0.189588	0.6	1
A.01.07.001.047	Marzipan pie	4	0.179281	0.6	1
A.01.07.002.006	Butter biscuits	4	0.016	0.6	1
A.05.08.003	Other fruit spreads	4	0.17	0.6	0.5
A.06.09.001.002	Bratwurst	4	0.059	1	1
A.06.09.001.004	Weisswurst	4	0.052	0.6	1
A.06.09.001.005	Bockwurst	4	0.052	0.6	1
A.06.11.001	Meat paste	4	0.075	0.6	1
A.06.11.005	Terrine	4	0.0458	0.6	1
A.07.02.002	Fishcakes	4	0.067	0.6	1
A.09	Egg products (unspecified)	4	1	0.6	1
A.09.01	Eggs, fresh (unspecified)	4	1	0.6	1
A.09.01.001	Whole egg, chicken	3	1	0.6	1
A.09.01.003	Egg white, chicken	3	1	1	1
A.09.01.004	Turkey eggs	3	1	0.6	1
A.09.01.005	Duck eggs	3	1	0.6	1
A.09.01.006	Goose eggs	3	1	0.6	1
A.09.01.007	Quail eggs	3	1	0.6	1
A.09.01.008	Ostrich eggs	3	1	0.6	1
A.09.01.009	Seagull eggs	3	1	0.6	1
A.09.02	Eggs, powder (unspecified)	4	1	0.6	1
A.09.02.001	Dried egg, whole	3	1.3	0.6	1
A.09.02.003	Dried egg white	3	1.3	1	1
A.10.04.010	Foamed sugar products (marshmallows)	3	0.1037	1	1
A.10.04.014	Nougat	3	0.0592	1	1
A.10.05.004	Alcoholic sweet sauce	3	0.2951	0.6	1
A.14.05.005	Egg liqueur	3	0.15	0.6	1
A.16.08	Savoury sauces (unspecified)	4	0.3692	0.6	1
A.19.01.001.006	Sandwich, cheese, egg and vegetable filling	4	0.033	0.6	1
A.19.01.002.002	Pizza and pizza‐like pies, cheese and vegetables	4	0.062	0.6	0.5
A.19.01.003	Pasta, cooked	4	0.078	0.6	0.5
A.19.01.003.001	Pasta, cooked, with vegetables	4	0.078	0.6	0.5
A.19.01.003.002	Pasta, cooked, with cheese/cream	4	0.078	0.6	0.5
A.19.01.003.003	Pasta, cooked, with vegetables and cheese/cream	4	0.078	0.6	0.5
A.19.01.003.004	Pasta, cooked, meat filling	4	0.078	0.6	0.5
A.19.01.003.005	Pasta, cooked, vegetable filling	4	0.078	0.6	0.5
A.19.01.003.006	Pasta, cooked, meat and vegetable filling	4	0.078	0.6	0.5
A.19.03.004	Potatoes and cheese meal	4	0.072	0.6	0.5
A.19.04.002	Beans and vegetables meal	4	0.065	0.6	0.5
A.19.05.001	Meat burger	3	0.0891	0.6	1
A.19.05.002	Meat balls	3	0.067	0.6	1
A.19.07.006	Vegetable soufflé	3	0.285	0.6	1
A.19.08	Egg‐based meal (unspecified)	4	0.6	0.6	1
A.19.08.001	Omelette, plain	3	0.961	0.6	1
A.19.08.002	Omelette with bacon	3	0.644	0.6	1
A.19.08.003	Omelette with cheese	3	0.853	0.6	1
A.19.08.004	Omelette with vegetables	3	0.644	0.6	1
A.19.08.005	Omelette with mushrooms	3	0.644	0.6	1
A.19.08.006	Fried eggs	3	0.9184	0.6	1
A.19.08.007	Poached eggs	3	1	0.6	1
A.19.10.007	Milk product/egg soup	3	0.1	0.6	1
A.19.11.010	Prepared mixed egg/meat/fish/vegetable salad	3	0.1	0.6	1
A.20.01.007	Fish‐based snacks	3	0.095	1	1
A.20.02.001	Ice cream, milk‐based	4	0.1006	0.6	1
A.20.02.003	Starchy pudding	3	0.059	0.6	1
A.20.02.004	Custard	4	0.1727	0.6	1

^(a)Source: Input data for the raw primary commodity model (available at https://doi.org/10.5281/zenodo.2537955).

^(b)Source: https://www.britannica.com/topic/egg-white.

Supporting information

Dietary exposure estimates to the food enzyme–TOS and the SMT–Equivalent in details

Suggested citation: EFSA CEP Panel (EFSA Panel on Food Contact Materials, Enzymes and Processing Aids) , Lambré C, Barat Baviera JM, Bolognesi C, Chesson A, Cocconcelli PS, Crebelli R, Gott DM, Grob K, Lampi E, Mengelers M, Mortensen A, Rivière G, Steffensen I‐L, Tlustos C, Van Loveren H, Vernis L, Zorn H, Andryszkiewicz M, Kovalkovicova N, Liu Y, di Piazza G, Ferreira de Sousa R and Chesson A, 2023. Scientific Opinion on the safety evaluation of the food enzyme lysozyme from hens' eggs. EFSA Journal 2023;21(3):7916, 17 pp. 10.2903/j.efsa.2023.7916