Updated safety evaluation of the food enzyme endo‐1,4‐β‐xylanase from the genetically modified Aspergillus niger strain XYL

TL;DR

This paper evaluates the safety of a food enzyme produced by a genetically modified fungus and concludes it is safe for use in food manufacturing.

Contribution

New genotoxicity data and a 90-day toxicity study confirm the safety of the food enzyme under intended use conditions.

Findings

Genotoxicity tests showed no safety concerns for the food enzyme.

The no observed adverse effect level was 4095 mg/kg bw/day for males and 4457 mg/kg bw/day for females.

The enzyme's amino acid sequence does not match known allergens, though a low risk of allergic reactions cannot be excluded.

Abstract

The food enzyme endo‐1,4‐β‐xylanase (4‐β‐d‐xylan xylanohydrolase, EC 3.2.1.8) is produced with the genetically modified Aspergillus niger strain XYL by DSM Food specialties. An evaluation of this food enzyme was made previously, in which EFSA could not conclude on its safety due to data gaps in a genotoxicity test. Subsequently, the applicant provided new data. The genetic modifications do not give rise to safety concerns. The food enzyme is free from viable cells of the production organism and its DNA. The food enzyme is intended to be used in four food manufacturing processes. Dietary exposure was estimated to be up to 0.281 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 4095 mg TOS/kg bw per day for males and of 4457 mg TOS/kg bw per day for females, respectively, the highest doses tested. When compared with the estimated dietary exposure, it results in a margin of exposure of at least 14,573. A search for the homology of the amino acid sequence of the endo‐1,4‐β‐xylanase to known allergens was made and no match was found. The Panel considered that a risk of allergic reactions upon dietary exposure to the food enzyme cannot be excluded, but the likelihood is low. Based on the new data and the data provided previously, the Panel concluded that this food enzyme does not give rise to safety concerns, under the intended conditions of use.

Tables

TABLE 1. Composition of the food enzyme.

Parameters	Unit	Batches
		1	2	3	4 a	5 b
Endo 1,4‐β‐xylanase activity	EDX/g c	57,550	47,965	44,050	193,300 d	49,400
Protein	%	19.4	17.7	17.2	76.2	18.3
Ash	%	0.7	0.4	0.5	2.0	0.6
Water	%	73.2	76.4	78.4	3.4	76.0
Total organic solids (TOS) e	%	26.1	23.2	21.1	94.6	23.4
Activity/TOS ratio	EDX/mg TOS	220	207	209	204	211

TABLE 2. Intended uses and recommended use levels of the food enzyme as provided by the applicant. 15

Food manufacturing process a	Raw material (RM)	Recommended use level (mg TOS/kg RM)
		Current evaluation b	Previous evaluation b , c
Processing of cereals and other grains
Production of baked products	Flour	0.14–3.40	0.16–1.17
Production of cereal‐based products other than baked	Flour	0.28–2.83
Processing of plant‐ and fungal‐derived products
Production of tea and other herbal and fruit infusions	Tea leaves and other plants (e.g. fruits, ginger, ginseng, etc.)	100–520
Production of plant extracts	Plant materials	100–520

TABLE 3. Summary of the 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)	0.003–0.066 (12)	0.021–0.097 (15)	0.026–0.085 (19)	0.012–0.044 (21)	0.008–0.043 (22)	0.006–0.052 (23)
Min–max 95th percentile (number of surveys)	0.010–0.195 (11)	0.057–0.281 (14)	0.063–0.220 (19)	0.029–0.105 (20)	0.018–0.125 (22)	0.016–0.106 (22)

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	+/−

Full text

INTRODUCTION

1

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 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.

Background and Terms of Reference as provided by the requestor

1.1

Background as provided by the European Commission

1.1.1

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 August 2022, a new application has been introduced by the applicant “DSM Food Specialties B.V.” for the authorisation of the food enzyme Xylanase from a genetically modified strain of Aspergillus niger (strain XYL).

Terms of Reference

1.1.2

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: Xylanase from a genetically modified strain of Aspergillus niger (strain XYL), in accordance with Regulation (EC) No 1331/2008 establishing a common authorisation procedure for food additives, food enzymes and food flavourings.

Interpretation of the Terms of Reference

1.2

The applicant previously submitted an application for xylanase from the genetically modified Aspergillus niger strain XYL, for which EFSA could not exclude safety concern of the food enzyme (EFSA CEF Panel, 2017).

The present opinion is a follow‐up of the previous assessment and assesses only new studies provided in the technical dossier. Whenever previous evaluation remains unchanged, reference to the previous EFSA opinion is made.

DATA AND METHODOLOGIES

2

Data

2.1

The applicant has submitted a dossier in support of the application for authorisation of the food enzyme endo‐1,4‐β‐xylanase from a genetically modified A. niger (strain XYL).

Additional information was requested from the applicant during the assessment process on 6 May 2024 and 18 October 2024 and received on 28 May 2024, and 17 December 2024 (see ‘Documentation provided to EFSA’).

Methodologies

2.2

The assessment was conducted in line with the principles described in the EFSA ‘Guidance on transparency in the scientific aspects of risk assessment’ (EFSA, 2009) 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 of the application.

Public consultation

2.3

According to Article 32c(2) of Regulation (EC) No 178/20023 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 15 June to 5 August 2024.4 No comments were received.

ASSESSMENT

3

An assessment of this food enzyme was made previously, in which EFSA could not conclude on its safety due to data gaps in a genotoxicity test (EFSA CEF Panel, 2017).

The applicant now provided new data and extended the intended uses to four food manufacturing processes: processing of cereals and other grains for the production of (1) baked products and (2) cereal‐based products other than baked; and processing of plant‐ and fungal‐derived products for the production of (3) tea and other herbal and fruit infusions and (4) plant extracts.IUBMB nomenclatureEndo 1,4‐β‐xylanaseSystematic name4‐β‐d‐xylan xylanohydrolaseSynonymsEndo‐(1–4)‐β‐xylan 4‐xylanohydrolase; xylanase; β‐1,4‐xylanase; β‐xylanaseIUBMB NoEC 3.2.1.8CAS No9025‐57‐4EINECS No232‐800‐2

Endo 1,4‐β‐xylanases catalyse the random hydrolysis of 1,4‐β‐d‐xylosidic linkages in xylans (including arabinoxylans) resulting in the generation of (1–4)‐β‐d‐xylan oligosaccharides of different lengths.

Source of the food enzyme

3.1

The enzyme is produced with the genetically modified filamentous fungus Aspergillus niger strain XYL (DS 26538), which is deposited at the Culture collection of the Westerdijk Fungal Biodiversity Institute (CBS, the Netherlands) with the deposition number ■■■■■.5 The production strain A. niger strain XYL was genetically modified to produce endo‐1,4‐β‐xylanase from A. tubingensis (formerly classified as A. niger). The strain was characterised in a previous scientific opinion (EFSA CEP Panel, 2017). The applicant has now provided a whole genome sequence (WGS) analysis of the production strain. Based on this, it was identified as A. niger, showing an average nucleotide identity of > 99% with the reference genome A. niger CBS 513.88.6 WGS analysis also revealed the presence of gene clusters for the biosynthesis of several secondary metabolites, suggesting that the production strain might have the capacity to produce these substances, none of them known to be toxic.7

WGS analysis of the production strain confirmed the ■■■■■ integration of ■■■■■ the expression cassette and the absence of vector backbone sequence including the antimicrobial resistance marker gene. No antimicrobial resistance genes were found by analysing the WGS against two maintained databases with thresholds of 80% identity and 70% coverage. Therefore, the Panel confirmed its previous conclusions regarding the safety of the genetic modifications.

Production of the food enzyme

3.2

See Sections 3.1.2 and 3.1.5 of the previous evaluation (EFSA CEF Panel, 2017). The applicant reported some minor changes in the manufacturing process that do not alter the chemical composition of the food enzyme.

Characteristics of the food enzyme

3.3

Properties of the food enzyme

3.3.1

See Sections 3.1.2 and 3.1.3 of the previous evaluation (EFSA CEF Panel, 2017).

Chemical parameters

3.3.2

Data on the chemical parameters of the food enzyme were provided for three batches used for commercialisation (batches 1–3) and for a batch produced for the repeated dose 90‐day oral toxicity study (batch 4) in the previous evaluation (EFSA CEF Panel, 2017). Data on the chemical parameters of the food enzyme were also provided for a new batch (batch 5), which was produced for the Ames test and for the in vitro mammalian cell micronucleus test 8 The mean total organic solids (TOS) of the three batches for commercialisation used in the previous evaluation (EFSA CEF Panel, 2017) was 23.5% and the mean enzyme activity/TOS ratio was 212 EDX/mg TOS.

Purity

3.3.3

Data on purity of the food enzyme for the three batches used for commercialisation (batches 1–3) and for the toxicological batch produced for the repeated dose 90‐day oral toxicity study (batch 4) were provided in the previous evaluation and found acceptable (EFSA CEF Panel, 2017).

Data on purity of the food enzyme were also provided for a new batch (batch 5) produced for the Ames test and for the in vitro mammalian cell micronucleus test. The lead content of batch 5 was below 5 mg/kg which complies with the specification for lead as laid down in the general specifications for enzymes used in food processing (FAO/WHO, 2006).

The batch 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). No antimicrobial activity was detected.

Strains of Aspergillus, in common with most filamentous fungi, have the capacity to produce a range of secondary metabolites (Frisvad et al., 2018). The presence of fumonisins B1, B2 and ochratoxin A, was examined and found below the limit of detection of the applied methods.

The Panel considered that the information provided on the purity of the newly evaluated batch is sufficient.

Viable cells and DNA of the production strain

3.3.4

The current evaluation supersedes section 3.3.4 of the previous evaluation (EFSA CEF Panel, 2017).

The absence of viable cells of the production strain was demonstrated in three independent batches of the food enzyme preparation analysed in triplicate. One gram of product was spread on selective agar plates and incubated at 30°C for 6 days. No colonies were observed. A positive control was included.10

The absence of recombinant DNA in the food enzyme was demonstrated by polymerase chain reaction (PCR) analysis of three batches in triplicate. No DNA was detected with primers that would amplify a 999‐bp fragment that is specific for the expression cassette, with a limit of detection of 10 ng spiked DNA/g food enzyme.11

Toxicological data

3.4

Batch 4 (Table 1) was used for the previously evaluated repeated dose 90‐day oral toxicity study in rats.

A new food enzyme batch was tested in the newly provided Ames test and an in vitro mammalian cell micronucleus test (Table 1, batch 5). This batch has a similar protein pattern and composition as the batches used for commercialisation, and thus is considered suitable as a test item.

Genotoxicity

3.4.1

The current evaluation supersedes Section 3.4.1 of the previous evaluation (EFSA CEF Panel, 2017).

Bacterial reverse mutation test

3.4.1.1

A bacterial reverse mutation test (Ames test) was performed according to the Organisation for Economic Co‐operation and Development (OECD) Test Guideline 471 (OECD, 1997) and following Good Laboratory Practice (GLP).12

Four strains of Salmonella Typhimurium (TA98, TA100, TA1535 and TA1537) and E. coli WP2uvrA were used with or without metabolic activation (S9‐mix), applying the standard plate incorporation method (dose‐range finding test and the first experiment) and pre‐incubation method (the second experiment).

The dose‐range finding test was carried out in triplicate, using eight concentrations of the food enzyme of 1.7, 5.4, 17, 52, 164, 512, 1600 and 5000 μg TOS/plate in S. Typhimurium TA100 and E. coli WP2uvrA in the absence and presence of S9‐mix. No precipitation and cytotoxicity were observed at any concentration of the test substance. No biologically relevant decrease in the number of revertants was observed.

The first experiment was carried out in triplicate, using five concentrations of the food enzyme of 52, 164, 512, 1600 and 5000 μg TOS/plate in S. Typhimurium TA1535, TA1537 and TA98 in the absence and presence of S9‐mix. No precipitation and cytotoxicity were observed at any concentration of the test substance. Upon treatment with the food enzyme, an increase in the number of revertant colonies was observed at concentrations of 52, 164 and 5000 μg TOS/plate and 52, 512 and 5000 μg TOS/plate in S. Typhimurium TA1537 without and with S9‐mix, respectively, however, without concentration response and within the historical control range. Therefore, this increase was considered not to be biologically relevant.

The second experiment was carried out in triplicate, using five concentrations of the food enzyme of 52, 164, 512, 1600 and 5000 μg TOS/plate in S. Typhimurium (TA98, TA100, TA1535 and TA1537) and E. coli WP2uvrA with or without S9‐mix. Slight precipitation was observed at the end of the incubation period at the concentration of 5000 μg TOS/plate in S. Typhimurium TA1535, TA1537 and E. coli WP2uvrA without S9‐mix. Toxic effects, evident as a reduction of the bacterial background lawn, occurred at 5000 μg TOS/plate in all strains without S9‐mix. Upon treatment with the food enzyme, an increase in the number of revertant colonies was observed at a concentration of 1600 μg TOS/plate in S. Typhimurium TA1535 without S9‐mix and at concentrations of 52 and 5000 μg TOS/plate in S. Typhimurium TA1537 with S‐9 mix, however, without concentration response and within the historical control range. Therefore, this increase was considered not to be biologically relevant.

The Panel concluded that the food enzyme endo‐1,4‐β‐xylanase did not induce gene mutations under the test conditions applied in this study.

In vitro mammalian cell micronucleus test

3.4.1.2

The in vitro mammalian cell micronucleus test was carried out according to the OECD Test Guideline 487 (OECD, 2016) and following GLP.13 Two separate 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).

In a range‐finding test, no cytotoxicity above 50% was seen at any concentration tested up to 2000 μg TOS/mL in the short‐term treatment with and without S9‐mix and in the long‐term treatment without S9‐mix.

In the first experiment, cells were exposed to the food enzyme and scored for the frequency of bi‐nucleated cells with micronuclei (MNBN) at concentrations of 500, 1000 and 2000 μg TOS/mL in a short‐term treatment (3 h exposure and 24‐h recovery period) either with or without S9‐mix. No cytotoxicity was observed. The frequency of MNBN was statistically significantly different to the negative controls at concentration of 1000 μg TOS/mL tested in the short‐term treatment without and with S9‐mix, however, without concentration response and within the 95% of the historical control range.

In the second experiment, cells were exposed to the food enzyme and scored for MNBN at concentrations of 500, 1000 and 2000 μg TOS/mL in a long‐term treatment (24 h exposure without recovery period) without S9‐mix. No cytotoxicity was seen. The frequency of MNBN was not statistically significantly different to the negative controls at all concentrations tested.

The Panel concluded that the food enzyme endo‐1,4‐β‐xylanase did not induce an increase in the frequency of MNBNs under the test conditions applied in this study.

Repeated dose 90‐day oral toxicity study in rodents

3.4.2

See Section 3.4.2 of the previous evaluation (EFSA CEF Panel, 2017). The Panel identified a no observed adverse effect level (NOAEL) at the highest dose tested of 4095 mg TOS/kg bw per day for males and 4457 mg TOS/kg bw per day for females (EFSA CEF Panel, 2017).

Allergenicity

3.4.3

The current evaluation supersedes Section 3.4.3 of the previous evaluation (EFSA CEF Panel, 2017).

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 endo 1,4‐β‐xylanase produced with the A. niger strain XYL 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 Allergen Online database.14

No reports on oral and respiratory sensitisation or elicitation reactions of the endo 1,4‐β‐xylanase under assessment have been published.

Respiratory allergy, e.g. baker's asthma, following occupational exposure to xylanase has been described in some epidemiological studies (Elms et al., 2003; Martel et al., 2010) and case reports (Baur et al., 1998; Lipińska‐Ojrzanowska et al., 2016; Merget et al., 2001). Several studies have shown that individuals respiratorily sensitised to a food enzyme are usually able to ingest the corresponding enzyme without acquiring clinical symptoms of food allergy (Armentia et al., 2009; Cullinan et al., 1997; Poulsen, 2004). Adverse reactions upon dietary exposure to xylanases in individuals sensitised through the respiratory route have not been reported.

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 endo‐1,4‐β‐xylanase under assessment.

The production strain belongs to the Aspergillus genus, which is known to cause respiratory allergy (Shen and 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.

Yeast extract, a known source of allergens, is present in the culture medium. During the fermentation process, this product 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 could be present in the food enzyme. Taking into account the level of dietary exposure (see Section 3.5.2), 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.

Dietary exposure

3.5

Intended use of the food enzyme

3.5.1

The current evaluation supersedes Sections 3.1.7 and 3.1.8 of the previous evaluation (EFSA CEF Panel, 2017).

The food enzyme is intended to be used in four 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. 15

For the production of baked products, the food enzyme is added to flour during the preparation of the dough or batter.16 It hydrolyses (arabino)xylans, which interact with gluten and bind water, reducing the dough viscosity and shortening the processing time. The decrease in viscosity also facilitates the handling of the dough and results in more uniform products. The food enzyme–TOS remain in the baked products.

For the production of cereal‐based products other than baked, the food enzyme is added to cereals and other grains during the preparation of the dough or batter.17 The endo‐1,4‐β‐xylanase hydrolyses (arabino)xylans, decreasing the water binding capacity of the dough and reducing viscosity. The food enzyme–TOS remain in the cereal‐based products.

In the production of tea and other herbal and fruit infusions, the food enzyme is added to the mash of tea leaves to degrade (arabino)xylans in the cell walls and consequently improve the extraction yield.18 The food enzyme–TOS remain in the final products.

In the production of plant extracts, obtained both with aqueous or organic solvent extraction,19 the food enzyme is added to the mash of plant materials to degrade (arabino)xylans in the cell walls and consequently improve the extractions yield.20 When used during the extraction with organic solvents, this endo‐1,4‐β‐xylanase reduces also the amount of solvent needed to obtain the extract.21 The food enzyme–TOS remain in the plant extracts.

Based on the thermostability data evaluated previously (EFSA CEF Panel, 2017), the Panel considered that this food enzyme is inactivated in all the food manufacturing processes listed in Table 2, with the exception of production of cereal‐based products other than baked, in which it may remain in its active form, depending on the drying conditions.

Dietary exposure estimation

3.5.2

The current evaluation supersedes sections 3.2.1, 3.2.2 and 3.2.3 of the previous evaluation (EFSA CEF Panel, 2017).

Chronic exposure to the food enzyme–TOS was calculated using the FEIM webtool22 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). 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 0.281 mg TOS/kg bw per day in toddlers at the 95th percentile.

Uncertainty analysis

3.5.3

The current evaluation supersedes section 3.2.4 of the previous evaluation (EFSA CEF Panel, 2017).

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.

The conservative approach applied to estimate the 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.

Margin of exposure

3.6

A comparison of the NOAEL (4095 mg TOS/kg bw per day in males) identified from the 90‐day rat study with the derived exposure estimates of 0.003–0.097 mg TOS/kg bw per day at the mean and 0.010–0.281 mg TOS/kg bw per day at the 95th percentile resulted in a margin of exposure of at least 14,573.

CONCLUSIONS

4

Based on the evaluation of the new data provided, the data previously submitted and the derived margin of exposure, the Panel concluded that the food enzyme endo 1,4‐β‐xylanase produced with the genetically modified Aspergillus niger strain XYL 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.

DOCUMENTATION AS PROVIDED TO EFSA

5

Application for the authorisation of endo‐1,4‐β‐xylanase from a genetically modified A. niger strain XYL as a new food enzyme. July 2023. Submitted by DSM Food Specialties B.V.

Additional data. May 2024. Submitted by DSM Food Specialties B.V.

Additional data. December 2024. Submitted by DSM Food Specialties B.V.ABBREVIATIONSbwbody weightCASChemical Abstracts ServiceCEFEFSA Panel on Food Contact Materials, Enzymes, Flavourings and Processing AidsCEPEFSA Panel on Food Contact Materials, Enzymes and Processing AidsEINECSEuropean Inventory of Existing Commercial Chemical SubstancesFAOFood and Agricultural Organization of the United NationsGLPGood Laboratory PracticeGMOgenetically modified organismIUBMBInternational Union of Biochemistry and Molecular BiologyMNBNbi‐nucleated cells with micronucleiNOAELno observed adverse effect levelOECDOrganisation for Economic Co‐operation and DevelopmentPCRpolymerase chain reactionTOStotal organic solidsWGSwhole genome sequencingWHOWorld Health Organization

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2023‐00522

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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 scientific output published implements EFSA's decision on the confidentiality requests submitted on specific items. As certain items have been awarded confidential status by EFSA they are consequently withheld from public disclosure by redaction.

Supporting information

Dietary exposure estimates to the food enzyme–TOS in details