Safety evaluation of the food enzyme pectin lyase from the genetically modified Aspergillus niger strain CCTCC M 2023341

Abstract

The food enzyme pectin lyase (1,4‐6‐O‐methyl‐α‐d‐galacturonan lyase, EC 4.2.2.10) is produced with the genetically modified Aspergillus niger strain CCTCC M 2023341 by Suntaq International Limited. The genetic modifications do not give rise to safety concerns. The food enzyme was considered free from viable cells of the production organism and its DNA. It is intended to be used in two food manufacturing processes. The dietary exposure was estimated to be up to 1.926 mg Total Organic Solids (TOS)/kg body weight (bw) per day in European populations. Genotoxicity tests did not indicate a safety concern. The systemic toxicity was assessed by means of a repeated dose 90‐day oral toxicity study in rats. The Panel identified a no observed adverse effect level of 2000 mg TOS/kg bw per day, the highest dose tested, which when compared with the estimated dietary exposure, resulted in a margin of exposure of at least 1038. A search for the homology of the amino acid sequence of the pectin lyase to known allergens was made and no match was found. The Panel considered that a risk of allergic reactions upon dietary exposure cannot be excluded, but that the likelihood is low. Based on the data provided, the Panel concluded that this food enzyme does not give rise to safety concerns, under the intended conditions of use.

1. INTRODUCTION

Article 3 of the Regulation (EC) No 1332/2008 ¹ 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 1331/2008 ² 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 EU 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.

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.

On 18 July 2024, a new application was introduced by the applicant “Suntaq International Limited” for the authorization of the food enzyme Pectin lyase produced from genetically modified Aspergillus niger (strain CCTCC M 2023341).

1.1.2. Terms of Reference

The European Commission requests the European Food Safety Authority to carry out the safety assessment and the assessment of possible confidentiality requests of the following food enzyme: Pectin lyase from genetically modified Aspergillus niger (strain CCTCC M 2023341) in accordance with Regulation (EC) No 1331/2008 establishing a common authorization procedure for food additives, food enzymes, and food flavorings.

2. DATA AND METHODOLOGIES

2.1. Data

The applicant has submitted a dossier in support of the application for authorisation of the food enzyme pectin lyase from A. niger CCTCC M 2023341.

Additional information was requested during the risk assessment phase. See https://open.efsa.europa.eu/questions/EFSA‐Q‐2024‐00525?search=2024‐00525.

Following the reception of additional data, EFSA requested a clarification teleconference on 15 June 2026, after which the applicant provided additional data on 16 June 2026.

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, 2009a) and following the relevant guidance documents of the EFSA Scientific Committee.

The ‘Scientific Guidance for the submission of dossiers on food enzymes’ (EFSA CEP Panel, 2021) and the ‘Food manufacturing processes and technical data used in the exposure assessment of food enzymes’ (EFSA CEP Panel, 2023) have been followed for the evaluation.

2.3. Public consultation

According to Article 32c(2) of Regulation (EC) No 178/2002 ³ and to the Decision of EFSA's Executive Director laying down the practical arrangements on pre‐submission phase and public consultations, EFSA carried out a public consultation on the non‐confidential version of the technical dossier from 23 April to 19 May 2025. ⁴ No comments were received.

3. ASSESSMENT

IUBMB nomenclature	Pectin lyase
Systematic name	(1‐4)‐6‐O‐methyl‐α‐d‐galacturonan lyase
Synonyms	Pectin trans‐eliminase; polymethylgalacturonic transeliminase; pectin methyltranseliminase
IUBMB no.	4.2.2.10
CAS no.	9033‐35‐6
EINECS no.	232‐894‐5

Pectin lyases catalyse the β‐eliminative cleavage of 1,4‐α‐d‐galactosiduronic linkages of pectin, resulting in the generation of oligosaccharides with 4‐deoxy‐6‐O‐methyl‐α‐d‐galacto‐4‐enuronosyl groups at their non‐reducing ends.

The food enzyme under assessment is intended to be used in two food manufacturing processes as defined in the EFSA guidance (EFSA CEP Panel, 2023): processing of fruits and vegetables for the production of (1) juices and (2) fruit and vegetable products other than juices.

3.1. Source of the food enzyme

The pectin lyase is produced with the genetically modified filamentous fungus Aspergillus. niger strain CCTCC M 2023341, which is deposited at the China Centre For Type Culture Collection (CCTCC, China) with deposition number M 2023341. ⁵ The production strain was identified as A. niger by whole genome sequence (WGS) analysis, which showed an average nucleotide identity (ANI) of 99.4% with ■■■■■ ⁶

3.1.1. Characteristics of the parental and recipient microorganisms

The parental strain is ■■■■■ ^{■■■■■} ■■■■■.

3.1.2. Characteristics of introduced sequences

The sequence encoding the pectin lyase ■■■■■.

■■■■■. ^{8 , 9}

3.1.3. Description of the genetic modification

The aim of the genetic modification was to enable the production strain to ■■■■■ pectin lyase enzyme.

The applicant stated that the presence and location of the enzyme encoding gene in the genome of the production strain were confirmed by ■■■■■. ¹⁰

3.1.4. Safety aspects of the genetic modification

The technical dossier contains all necessary information on the recipient microorganism, the donor organism and the genetic modification process.

The production strain A. niger CCTCC M 2023341 differs from the recipient strain in its capacity to ■■■■■ pectin lyase and its ■■■■■.

No issues of concern arising from the genetic modifications were identified by the Panel.

3.2. Production of the food enzyme

The food enzyme is manufactured according to the Food Hygiene Regulation (EC) No 852/2004, ¹¹ with food safety procedures based on Hazard Analysis and Critical Control Points and in accordance with Good Manufacturing Practice.

The production strain is grown as a pure culture using a typical industrial medium in a submerged, fed‐batch fermentation system with conventional process controls in place. After completion of the fermentation, the solid biomass is removed from the fermentation broth by filtration. The filtrate containing the enzyme is then further purified and concentrated, including an ultrafiltration step in which enzyme protein is retained, while most of the low molecular mass material passes the filtration membrane and is discarded. ¹² The applicant provided information on the identity of the substances used to control the fermentation and in the subsequent downstream processing of the food enzyme. ¹³

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

The pectin lyase is composed of ■■■■■ amino acids ¹⁴ and the molecular mass of the mature protein, calculated from the amino acid sequence, is ■■■■■ kDa. ¹⁵ The food enzyme was analysed by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. ¹⁶ A consistent protein pattern was observed across all batches. The gel showed a protein band migrating around the ■■■■■ kDa marker, consistent with the expected molecular mass of the enzyme. Another protein band was observed between ■■■■■ and ■■■■■ kDa, identified by the applicant as a polygalacturonase.

Polygalacturonase is reported as side activity. ¹⁷ No other enzyme activities were reported.

The determination of pectin lyase activity is based on the eliminative cleavage of pectin (reaction conditions: pH 3.5, 40°C, 10 min). The degree of unsaturation of the released pectin oligomers is measured spectrophotometrically at 235 nm.

The enzyme activity is expressed in Pectin lyase units (U)/g. One unit is defined as the amount of food enzyme that increases the absorbance of the reaction solution by 0.1 per min. ¹⁸

The food enzyme has a temperature optimum around ■■■■■ (■■■■■) and a pH optimum around ■■■■■ (■■■■■). Thermostability was tested by pre‐incubation of the food enzyme for ■■■■■. The enzyme activity decreased above ■■■■■ showing no residual activity at ■■■■■. ¹⁹

3.3.2. Chemical parameters

Data were provided for three batches of the food enzyme intended for commercialisation and one batch produced for the toxicological tests (Table 1). ²⁰ The mean Total Organic Solids (TOS) of the three batches intended for commercialisation was 22.3% and the mean enzyme activity/TOS ratio was 165.0 U/mg TOS.

TABLE 1.

Composition of the food enzyme.

Parameters	Unit	Batches
		1	2	3	4 a
Pectin lyase activity	U/g b	35,100	37,710	38,097	30,250
Protein	%	15.8	17.1	17.1	13.8
Ash	%	1.8	2.1	1.8	3.6
Water	%	76.6	75.4	75.4	6.6
Total Organic Solids (TOS) c	%	21.6	22.5	22.8	88.8
Pectin lyase activity/TOS ratio	U/mg TOS	162.5	167.6	167.1	33.7

^a Batch used for the toxicological studies.

^b U: Pectin lyase activity unit (see Section 3.3.1).

^c TOS calculated as 100% – % water – % ash.

3.3.3. Purity

The lead content in all batches was below 0.05 mg/kg ²¹ 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, cadmium and mercury concentrations were measured in the four batches and were below the limits of quantification (LoQ) of the employed methods. For arsenic the highest concentration determined in the three commercial batches was 0.0733 mg/kg. ^{22 , 23} The Panel considered this concentration as not of concern.

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). ^{24 , 25} No antimicrobial activity was detected in any of the tested batches. ²⁶

Strains of Aspergillus species in common with most filamentous fungi have the capacity to produce a range of secondary metabolites (Frisvad et al., 2018). The presence of aflatoxins B1, B2, G1 and G2, deoxynivalenol, fumonisins B1 and B2, ochratoxin A, sterigmatocystin, HT‐2 toxin, T‐2 toxin and zearalenone was examined in all food enzyme batches and all were below the LoQ of the applied methods. ^{27 , 28 , 29} Adverse effects caused by the possible presence of other secondary metabolites are addressed by the toxicological examination of the food enzyme.

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

3.3.4. Viable cells and DNA of the production strain

The absence of viable cells of the production strain in the food enzyme was demonstrated in three independent batches analysed in triplicate. ■■■■■. No colonies were detected. ³⁰ A positive control was used.

The absence of recombinant DNA in the food enzyme was demonstrated by polymerase chain reaction analysis of three batches in triplicate. No DNA was detected with primers that would amplify ■■■■■, with a limit of detection of 10 ng spiked DNA/g food enzyme. ^{31 , 32}

3.4. Toxicological data

A battery of toxicological tests including a bacterial reverse mutation test (Ames test), an in vitro mammalian cell micronucleus test and a repeated dose 90‐day oral toxicity study in rats has been provided.

The batch 4 (Table 1) used in these studies has a lower enzyme activity/TOS ratio compared to the batches intended for commercialisation and thus it is considered suitable as a test item.

3.4.1. Genotoxicity

3.4.1.1. Bacterial reverse mutation test

A bacterial reverse mutation test (Ames test) was performed according to the Organisation for Economic Co‐operation and Development (OECD) Test Guideline 471 (OECD, 2020) and following Good Laboratory Practice (GLP). ³³ Five strains of Salmonella Typhimurium (TA98, TA100, TA102, TA1535 and TA1537) were used, with or without metabolic activation (S9‐mix), applying the pre‐incubation and the standard plate incorporation methods. A dose‐finding test in duplicate and two main experiments in triplicate were conducted.

Based on the results from the dose‐finding test, the two main experiments were carried out, using five concentrations of the food enzyme of 58, 158, 500, 1580 and 5000 μg TOS/plate.

No cytotoxicity was observed at any concentration of the test item. Upon treatment with the food enzyme there was no biologically relevant increase in the number of revertant colonies above the control values, in any strain tested, with or without S9‐mix.

The Panel concluded that the food enzyme did not induce gene mutations under the test conditions applied in this study.

3.4.1.2. In vitro mammalian cell micronucleus test

The in vitro mammalian cell micronucleus test was carried out according to the OECD Test Guideline 487 (OECD, 2023) and following GLP. ³⁴ A dose‐finding test and two main experiments were performed with duplicate cultures of human peripheral whole blood lymphocytes. The cell cultures were treated with the food enzyme with or without metabolic activation (S9‐mix).

Based on the results of the dose‐finding test, in the first main experiment, cells were exposed to the food enzyme and scored for the frequency of bi‐nucleated cells with micronuclei (MNBN) at concentrations of 1250, 2500 and 5000 μg TOS/mL in a short‐term treatment (4‐h exposure and 19‐h recovery period) either with or without S9‐mix.

In the second main experiment, cells were exposed to the food enzyme and scored for the frequency of MNBN at concentrations of 1250, 2500 and 5000 μg TOS/mL in a long‐term treatment (24‐h exposure and 19‐h recovery period) without S9‐mix.

No cytotoxicity was seen either in the short‐term treatment with and without S9‐mix, or in the long‐term treatment. The frequency of MNBN was not statistically significantly different to the negative controls at any concentration tested.

The Panel concluded that the food enzyme did not induce an increase in the frequency of MNBN under the test conditions applied in this study.

3.4.2. Repeated dose 90‐day oral toxicity study in rodents

The repeated dose 90‐day oral toxicity study was performed under GLP and according to the OECD Test Guideline 408 (OECD, 2018) ³⁵ with the following deviation: plasmatic urea was not measured. The Panel considered that this deviation is minor and does not impact the evaluation of the study.

Groups of 10 male and 10 female Sprague–Dawley rats (Crl:CD(SD)) received the food enzyme by gavage in doses of 500, 1000 or 2000 mg TOS/kg body weight (bw) per day. Controls received the vehicle (Ultrapure water).

No mortality was observed.

The feed consumption was statistically significantly increased in week 9 of administration in low‐dose females (+9%). The Panel considered the change as not toxicologically relevant, as it was only recorded at a single time interval, it was only observed in one sex, there was no dose–response relationship and there was no statistically significant change in the final feed consumption, in the body weight and the body weight gain.

Haematological investigations revealed a statistically significant increase in neutrophil (Neu) and lymphocyte (Lymph) counts in mid‐dose males (+52% both) resulting in an increase in total white blood cells (WBC; +51%). An increase in Neu was also observed in mid‐dose females (+2%). The Panel considered the changes as not toxicologically relevant, as they were only observed in one sex (Lymph, WBC), there was no dose‐response relationship (all parameters), there were no changes in other relevant parameters (other WBC), there were no histopathological changes (no inflammation in any tissue/organ examined) and the changes were within the historical control values.

Clinical chemistry investigations revealed a statistically significant increase in calcium concentration (Ca) in all treated male groups (+3%, +3%, +4%), a decrease in sodium concentration (Na) in all treated female groups (−1%, −2%, −1%) and in mid‐dose males (−2%), an increase in total protein (TP) and globulin (Glo) in high‐dose males (+8% both), an increase in glucose (Glc) and blood urea nitrogen (BUN) concentrations in low‐dose females (+20%, +23%) and an increase in high and low‐density lipoproteins in high‐dose females (+24%, +50%). The Panel considered the changes as not toxicologically relevant, as they were only observed in one sex (all except Na), there was no dose–response relationship (Ca, Na, Glc, BUN), the changes were small (TP and Glo), there were no histopathological changes in the pancreas (Glc) or kidneys (BUN) and all the changes were within the historical control values.

Statistically significant changes in hormone levels included a decrease in tri‐iodothyronine (T3) in mid‐ and high‐dose females (−39%, −42%). The Panel considered this difference as not test item related because there was no evidence of disruption of the hypothalamic–pituitary‐thyroid axis (i.e. T4, TSH, thyroid histopathology).

Statistically significant changes detected in organ weights were an increase in absolute and relative to body weight thyroid and parathyroid weights in high‐dose females (+44%, +51%). The Panel considered the changes as not toxicologically relevant, as they were only observed in one sex (both parameters); the change was not compatible with the decrease in T3 observed in mid‐ and high‐dose females, and there were no histopathological changes in thyroid or pituitary glands.

No toxicologically relevant or other statistically significant differences from controls were reported.

The Panel identified a no observed adverse effect level (NOAEL) of 2000 mg TOS/kg bw per day, the highest dose tested.

3.4.3. Allergenicity

The allergenicity assessment considered only the food enzyme and not additives, carriers or other excipients that may be used in the final formulation.

The potential allergenicity of the pectin lyase produced with the filamentous fungus A. niger strain CCTCC M 2023341 was assessed by comparing its amino acid sequence with those of known allergens as described in the EFSA GMO Scientific Opinion (EFSA, GMO Panel 2010). Using higher than 35% identity in a sliding window of 80 amino acids as the criterion, no match was found in the AllergenOnline database. ³⁶

No reports on oral or respiratory sensitisation or elicitation reactions of the food enzyme under assessment have been published. In addition, no allergic reactions to any pectin lyase have been reported. ³⁷

The Panel considered that the results of the sequence homology search and the available literature do not indicate a risk of allergic reactions upon dietary exposure to the food enzyme under assessment.

The production strain belongs to the Aspergillus genus, which is known to cause respiratory allergy (Kurup et al., 2000; Shen & Han, 1998; Vermani et al., 2015). Allergic reactions upon dietary exposure have been observed, but are rare (Xing et al., 2022). The biomass is removed during the production process; however, allergenic proteins of the production strain can be released into the culture medium from which the food enzyme is obtained.

■■■■■ and ■■■■■, known sources of allergens, are present in the culture medium. During the fermentation process, these products will mostly be degraded and utilised by the production strain.

Taken together, concerning the potential allergic reactions due to the production strain and the raw material in the culture medium, the Panel considered that residual amounts of allergenic proteins are present in the food enzyme. Taking into account the level of dietary exposure (see Section 3.5), this would result in minute amounts in the final foods, from which allergic reactions are usually not expected.

In conclusion, the Panel considered that under the conditions of use, a risk of allergic reactions upon dietary exposure to this food enzyme cannot be excluded, but that the likelihood is low.

3.5. Dietary exposure

3.5.1. Intended use of the food enzyme

The food enzyme is intended to be used in two food manufacturing processes at the recommended use levels summarised in Table 2.

TABLE 2.

Intended uses and recommended use levels of the food enzyme as provided by the applicant. ³⁸

Food manufacturing process	Raw material (RM)	Maximum recommended use level (mg TOS/kg RM) a
Processing of fruits and vegetables
Production of juices	Fruit and vegetables	44.6
Production of fruit and vegetable products other than juices	Fruit and vegetables	44.6

Abbreviation: TOS, Total Organic Solids.

^a The numbers in bold were used for calculation.

In all the intended food manufacturing processes, the food enzyme is used to treat plant materials to catalyse the degradation of pectin, thereby improving the processability and the yields.

In the production of juices, the food enzyme may be added to fruit and vegetables before maceration or to the raw juice during the depectinisation. ³⁹ The food enzyme–TOS remain in the juices.

In the production of fruit and vegetable products other than juices, the food enzyme is added to fruit and vegetables after the crushing. ⁴⁰ The food enzyme–TOS remain in the final foods.

Based on data provided on thermostability (see Section 3.3.1) and the downstream processing within the respective food manufacturing processes, the Panel considered that the food enzyme may remain in its active form in the food manufacturing processes listed in Table 2, depending on the heat treatment conditions.

3.5.2. Dietary exposure estimation

Chronic exposure to the food enzyme–TOS was calculated using the FEIM webtool ⁴¹ by combining the maximum recommended use level with individual consumption data (EFSA CEP Panel, 2021). The estimation involved selection of relevant food categories and application of technical conversion factors (EFSA CEP Panel, 2023).

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 51 dietary surveys (covering infants, toddlers, children, adolescents, adults and the elderly), carried out in 27 European countries (Appendix B). The highest dietary exposure was estimated to be 1.926 mg TOS/kg bw per day in children at the 95th percentile.

TABLE 3.

Summary of the estimated dietary exposure to the 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	4–11 months	12–35 months	3–9 years	10–17 years	18–64 years	≥ 65 years
Min–max mean (number of surveys)	0.043–0.632 (14)	0.173–1.233 (17)	0.066–0.695 (21)	0.042–0.387 (23)	0.023–0.243 (23)	0.008–0.159 (25)
Min–max 95th percentile (number of surveys)	0.172–1.260 (13)	0.634–1.861 (16)	0.209–1.926 (21)	0.143–1.195 (22)	0.085–0.896 (23)	0.035–0.633 (24)

Abbreviation: TOS, Total Organic Solids.

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 4.

TABLE 4.

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

Sources of uncertainties	Direction of impact
Model input data
Consumption data: different methodologies/representativeness/ underreporting/misreporting/no portion size standard	+/−
Use of data from food consumption surveys 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
Selection of broad FoodEx categories for the exposure assessment	+
Exposure to food enzyme–TOS always calculated based on the recommended maximum use level	+
Use of recipe fractions to disaggregate FoodEx categories	+/−
Use of technical factors in the exposure model	+/−

Note: +: uncertainty with potential to cause overestimation of exposure. –: uncertainty with potential to cause underestimation of exposure.

Abbreviation: TOS, Total Organic Solids.

The conservative approach applied to estimate the dietary exposure to the food enzyme–TOS, in particular assumptions made on the occurrence and use levels of this specific food enzyme, is likely to have led to an overestimation of the exposure.

3.6. Margin of exposure

A comparison of the NOAEL (2000 mg TOS/kg bw per day) identified from the 90‐day rat study with the derived exposure estimates of 0.008–1.233 mg TOS/kg bw per day at the mean and 0.035–1.926 mg TOS/kg bw per day at the 95th percentile resulted in a margin of exposure of at least 1038.

4. CONCLUSIONS

Based on the data provided and the derived margin of exposure, the Panel concluded that the food enzyme pectin lyase produced with the genetically modified A. niger strain CCTCC M 2023341 does not give rise to safety concerns under the intended conditions of use.

The Panel considered the food enzyme free from viable cells of the production organism and recombinant DNA.

5. DOCUMENTATION AS PROVIDED TO EFSA

Dossier and additional information can be accessed at https://open.efsa.europa.eu/dossier/FEN‐2024‐28032.

ABBREVIATIONS

AMR

antimicrobial resistance

BUN

blood urea nitrogen

bw

body weight

Ca

calcium

CAS

Chemical Abstracts Service

CCTCC

China Centre for Type Culture Collection

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

FEZ

EFSA Panel on Food Enzymes

Glo

globulin

GLP

Good Laboratory Practice

Glu

glucose

GMM

genetically modified microorganism

GMO

genetically modified organism

IUBMB

International Union of Biochemistry and Molecular Biology

JECFA

Joint FAO/WHO Expert Committee on Food Additives

kDa

kiloDalton

LOQ

limit of quantification

Lymph

lymphocyte

MNBN

bi‐nucleated cells with micronuclei

Na

sodium

Neu

neutrophil

OECD

Organisation for Economic Co‐operation and Development

PCR

polymerase chain reaction

SDS‐PAGE

sodium dodecyl sulfate‐polyacrylamide gel electrophoresis

TOS

total organic solids

TP

total protein

WGS

whole genome sequencing

WHO

World Health Organization

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2024‐00525

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PANEL MEMBERS

José Manuel Barat Baviera, Claudia Bolognesi, Francesco Catania, Gabriele Gadermaier, Ralf Greiner, Baltasar Mayo, Alicja Mortensen, Yrjö Henrik Roos, Marize de Lourdes Marzo Solano, Monika Sramkova, Henk Van Loveren, Laurence Vernis and Holger Zorn.

LEGAL NOTICE

The published scientific output includes information items that have been redacted, in accordance with the confidentiality requests submitted by the applicant, pending a decision thereon by EFSA. Redactions may be subject to review once a decision on such confidentiality requests is taken by EFSA.

Supporting information

APPENDIX A Dietary exposure estimates to the food enzyme–TOS in details

APPENDIX A. Dietary exposure estimates to the food enzyme–TOS in details

Appendix A can be found in the online version of this output (in the ‘Supporting Information’ section). The file contains two sheets, corresponding to two tables.

Table 1: Average and 95th percentile exposure to the food enzyme–TOS per age class, country and survey.

Table 2: Contribution of food categories to the dietary exposure to the food enzyme–TOS per age class, country and survey.

APPENDIX B. Population groups considered for the exposure assessment

Population	Age range	Countries with food consumption surveys covering more than 1 day
Infants	From 16 weeks on up to and including 11 months of age	Bulgaria, Croatia, Cyprus, Denmark, Estonia, Finland, France, Germany, Italy, Latvia, Poland, Portugal, Slovenia, Spain
Toddlers	From 12 months up to and including 35 months of age	Belgium, Bulgaria, Croatia, Cyprus, Denmark, Estonia, Finland, France, Germany, Hungary, Italy, Latvia, Montenegro,* the Netherlands, Poland, Portugal, Republic of North Macedonia,* Serbia,* Slovenia, Spain
Children	From 36 months up to and including 9 years of age	Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Italy, Latvia, Montenegro,* the Netherlands, Poland, 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,* Croatia, Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Italy, Latvia, Montenegro,* the Netherlands, Poland, 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,* the Netherlands, Poland, Portugal, Romania, Serbia,* Slovenia, Spain, Sweden
The elderly a	From 65 years of age and older	Austria, Belgium, Croatia, Cyprus, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Ireland, Italy, Latvia, Montenegro,* the Netherlands, Poland, Portugal, Romania, Serbia,* Slovenia, Spain, Sweden

^*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.

^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).

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