Safety evaluation of the food enzyme β‐glucosidase from the non‐genetically modified Penicillium guanacastense strain AE‐GLY

TL;DR

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

Contribution

A safety evaluation of β-glucosidase from Penicillium guanacastense for food use, including toxicity and allergenicity assessments.

Findings

Genotoxicity tests showed no safety concerns for the food enzyme.

The no observed adverse effect level was 943 mg TOS/kg bw per day, with a margin of exposure of at least 233.

No sequence similarity to known allergens was found, though a low risk of allergic reactions cannot be excluded.

Abstract

The food enzyme β‐glucosidase (β‐D‐glucoside glucohydrolase; EC 3.2.1.21) is produced with the non‐genetically modified Penicillium guanacastense strain AE‐GLY by Amano Enzyme Inc. The food enzyme is intended to be used in four food manufacturing processes. Dietary exposure to the food enzyme‐total organic solids (TOS) was estimated to be up to 4.054 mg TOS/kg body weight (bw) per day in European populations. Genotoxicity tests did not raise 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 943 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 233. A search for the similarity of the amino acid sequence of the food enzyme to known allergens was made and no match was found. The Panel considered that the risk of allergic reactions by dietary exposure cannot be excluded, but 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.

Tables

TABLE 1. Composition of the food enzyme preparation.

Parameters	Unit	Batches
		1	2	3	4 a
β‐Glucosidase activity	U/g b	2210	2290	2330	1025
Protein	%	41.1	43.1	44.9	44.0
Ash	%	1.3	1.3	0.9	1.0
Water	%	6.4	6.1	5.9	4.7
■■■■■ (diluent)	%	38.1	36.8	34.4	0
Total organic solids (TOS) c	%	54.2	55.8	58.8	94.3
Activity/ TOS ratio	U/mg TOS	4.1	4.1	4.0	1.1

TABLE 2. Intended uses and use levels of the food enzyme as provided by the applicant. 22

Food manufacturing process a	Raw material (RM)	Recommended use level (mg TOS/kg RM) b
Processing of fruit and vegetables
Production of juices	Citrus fruit 23	18.9–94.5
Production of wine and wine vinegar	Grapes	2.3–23
Processing of plant‐ and fungal‐derived products
Production of tea and other herbal and fruit infusions	Tea	0.5–927
Production of plant‐based analogues of milk and milk products	Soy drink	9.3–46
	Cereals 24	4.1–82.2

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)	0.029–0.636 (12)	0.170–2.300 (15)	0.008–1.298 (19)	0.012–0.734 (21)	0.050–0.477 (22)	0.036–0.325 (23)
Min–max 95th percentile (number of surveys)	0.106–2.396 (11)	1.015–3.896 (14)	0.029–4.054 (19)	0.054–2.429 (20)	0.242–1.810 (22)	0.145–1.229 (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
Exposure to food enzyme–TOS was always calculated based on the recommended maximum use level	+
Selection of broad FoodEx categories for the exposure assessment	+
For the production of juices, the calculation considered all fruits, although the applicant specified only citrus fruit.	+
Use of recipe fractions in disaggregation FoodEx categories	+/−
Use of technical factors in the exposure model	+/−

Full text

INTRODUCTION

1

Article 3 of the Regulation (EC) No 1332/2008 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 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 European Union (EU) Community 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.

Five applications have been introduced by the companies ‘Amano Enzyme Inc.’, ‘DSM Food Specialties B.V.’ and ‘Novozymes A/S’ for the authorization of food enzymes Glucoamylase from Rhizopus oryzae (strain AE‐G), Beta‐glucosidase from Penicillium multicolour (strain AE‐GLY), Peroxidase from a genetically modified strain of Aspergillus niger (strain MOX), Beta‐amylase from a genetically modified strain of Bacillus licheniformis (strain NZYM‐JA) and Triacylglycerol lipase from Aspergillus niger (strain AE‐L), respectively.

Following the requirements of Article 12.1 of Regulation (EC) No 234/20111 implementing Regulation (EC) No 1331/2008, the Commission has verified that the five applications fall within the scope of the food enzyme Regulation and contain all the elements required under Chapter II of that Regulation.

Terms of Reference

1.1.2

The European Commission requests the European Food Safety Authority to carry out the safety assessments of the food enzymes Glucoamylase from Rhizopus oryzae strain AE‐G, Beta‐glucosidase from Penicillium multicolor strain AE‐GLY, Peroxidase from a genetically modified strain of Aspergillus niger strain MOX, Beta‐amylase from a genetically modified strain of Bacillus licheniformis strain NZYM‐JA and Triacylglycerol lipase Aspergillus niger strain AE‐L in accordance with Article 17.3 of Regulation (EC) No 1332/2008 on food enzymes.

Interpretation of the Terms of Reference

1.2

The present scientific opinion addresses the European Commission's request to carry out the safety assessment of food enzyme beta‐glucosidase from Penicillium multicolor strain AE‐GLY.

Although the original mandate refers the food enzyme as produced from Penicillium multicolor strain AE‐GLY, the new data package identified the production microorganism as Penicillium guanacastense strain AE‐GLY.

DATA AND METHODOLOGIES

2

Data

2.1

The applicant has submitted a dossier in support of the application for authorisation of the food enzyme beta‐glucosidase from Penicillium multicolor strain AE‐GLY. The dossier was updated on 21 October 2015.

Additional information was requested from the applicant during the assessment process on 29 September 2021 and received on 28 February 2022 (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, 2009a) and following the relevant guidance documents of the EFSA Scientific Committee.

The ‘Guidance on the submission of a dossier on food enzymes for safety evaluation’ (EFSA, 2009b) have been followed for the evaluation of the application. Additional information was requested in accordance with the updated ‘Scientific Guidance for the submission of dossiers on food enzymes’ (EFSA CEP Panel, 2021) and the guidance on the ‘Food manufacturing processes and technical data used in the exposure assessment of food enzymes’ (EFSA CEP Panel, 2023).

ASSESSMENT

3

IUBMB nomenclatureβ‐GlucosidaseSystematic nameβ‐D‐glucoside glucohydrolaseSynonymsβ‐D‐glucosidase, cellobiase, emulsinIUBMB NoEC 3.2.1.21CAS No9001‐22‐3EINECS No232‐589‐7

β‐Glucosidases catalyse the hydrolysis of (1,4)‐β‐D‐glucosidic linkages in β‐D‐glucans, releasing β‐D‐glucose. The food enzyme under assessment is intended to be used in four food manufacturing processes as described in the EFSA Guidance (EFSA CEP Panel, 2023): processing of fruits and vegetables for the production of (1) juices and (2) wine and wine vinegar and processing of plant‐ and fungal‐derived products for the production of (3) tea and other herbal and fruit infusions and (4) plant‐based analogues of milk and milk products.

Source of the food enzyme

3.1

The β‐glucosidase is produced with the non‐genetically modified filamentous fungus Penicillium guanacastense strain AE‐GLY, which is deposited at the National Institute of Technology and Evaluation (NITE) Biological Resource Center (Japan), with the deposit number ■■■■■. The production strain was identified as P. guanacastense by a ■■■■■.

The parental strain was deposited as ■■■■■. The production strain P. guanacastense AE‐GLY was derived from the parental strain ■■■■■.

Production of the food enzyme

3.2

The food enzyme is manufactured according to the Food Hygiene Regulation (EC) No 852/2004,2 with food safety procedures based on Hazard Analysis and Critical Control Points, and in accordance with current good manufacturing practice.

The production strain is grown as a pure culture using a typical industrial medium in a submerged, batch or 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, leaving a filtrate containing the food enzyme. 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.3 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.4

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.

Characteristics of the food enzyme

3.3

Properties of the food enzyme

3.3.1

The β‐glucosidase is a single polypeptide chain of ■■■■■ amino acids.5 The molecular mass of the mature protein, calculated from the amino acid sequence, is ■■■■■ kDa. The food enzyme was analysed by size exclusion chromatography.6 The chromatograms of the three food enzyme batches for commercialisation showed similar patterns. No other enzyme activities were reported.7

The in‐house determination of β‐glucosidase activity is based on the hydrolysis of p‐nitrophenyl‐β‐D‐glucopyranoside (reaction conditions: pH ■■■■■, ■■■■■C, ■■■■■ min) and determined by measuring the release of p‐nitrophenol spectrophotometrically at 412 nm. The β‐glucosidase activity is expressed in Unit/g. One unit is defined as the amount of enzyme required to release 1 μmol of p‐nitrophenol per minute under the conditions of the assay.8

The food enzyme has a temperature optimum between 60°C and 70°C (pH 4.5) and a pH optimum between pH 4.5 and 5.0 (37°C). Thermostability was tested after a pre‐incubation of the food enzyme for 30 min at different temperatures (pH 4.5). Enzyme activity decreased above 60°C, showing no residual activity after pretreatment above 80°C.9

Chemical parameters

3.3.2

Data on the chemical parameters of the food enzyme preparation were provided for three batches used for commercialisation and one batch produced for the toxicological tests (Table 1).10 The mean total organic solids (TOS) of the three food enzyme batches for commercialisation was 56.3% and the mean enzyme activity/TOS ratio was 4.1 U/mg TOS.

Purity

3.3.3

The lead content in the three commercial batches was equal to or below 0.014 mg/kg11 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, concentrations of cadmium and mercury were below the limits of quantification (LoQ) of the employed method. For arsenic, the average concentration determined in the commercial batches was 0.34 mg/kg.12 ^,^ 13 The Panel considered this concentration as of no concern.

The food enzyme preparation 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 No antimicrobial activity was detected in any of the tested batches.15

Strains of Penicillium, in common with most filamentous fungi, have the capacity to produce a range of secondary metabolites. The presence of aflatoxins, deoxynivalenol, HT‐2 toxin, T‐2 toxin, zearalenone, ochratoxin A and sterigmatocystin was examined in the three food enzyme batches. All were below the LoQs of the applied analytical methods.16 Adverse effects caused by the possible presence of other secondary metabolites are addressed by the toxicological examination of the food enzyme TOS.

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

Viable cells of the production strain

3.3.4

The absence of viable cells of the production strain in the food enzyme was demonstrated ■■■■■.

Toxicological data

3.4

A battery of toxicological tests, including a bacterial gene mutation assay (Ames test), an in vitro mammalian chromosomal aberration 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 lower activity/TOS value than the batches intended for commercialisation and was considered suitable as a test item.

Genotoxicity

3.4.1

Bacterial reverse mutation test

3.4.1.1

A bacterial reverse mutation assay (Ames test) was performed according to Organisation for Economic Co‐operation and Development (OECD) Test Guideline 471 (OECD, 1997a) and following good laboratory practice (GLP).17

Four strains of Salmonella Typhimurium (TA98, TA100, TA1535 and TA1537) and Escherichia coli WP2 uvrA were used in the presence or absence of metabolic activation (S9‐mix), applying the pre‐incubation method. Based on the results of a concentration range‐finding test, the main experiments were carried out using six concentrations of the food enzyme (from 156 to 5000 μg dry matter/plate, corresponding to 147, 295, 589, 1179, 2358 and 4716 μg TOS/plate), with triplicate plating. No cytotoxicity was observed at any concentration of the food enzyme under any of the test conditions. Upon treatment with the food enzyme, there was no significant increase in revertant colony numbers above the control values in any strain or at any concentration of the food enzyme tested.

The Panel concluded that the food enzyme β‐glucosidase did not induce gene mutations under the test conditions employed in this study.

In vitro mammalian chromosomal aberration test

3.4.1.2

The in vitro mammalian chromosomal aberration test was carried out with Chinese hamster lung fibroblast cells according to OECD Test Guideline 473 (OECD, 1997b) and following GLP.18

The range‐finding study was performed at eight concentrations ranging from 39.1 to 5000 μg/mL in a short‐term treatment (6 hours +18 hours of recovery period) with and without metabolic activation (S9‐mix) and in 24 and 48 hours of continuous treatments without S9‐mix. No inhibition of cell growth of 50% was observed at any of the test conditions. In the main experiment, chromosomal aberrations were scored in duplicate cultures at concentrations of 625, 1250, 2500 and 5000 μg/mL (corresponding to 589, 1179, 2358 and 4716 μg TOS/mL), both in the short‐term treatment with and without S9‐mix, and in the continuous treatments (24 hours or 48 hours) in the absence of S9‐mix.

The highest cytotoxicity (30%) was observed at 5000 μg/mL in the 48‐hour treatment without S9‐mix. The frequency of structural and numerical chromosomal aberrations in treated cultures was comparable to those seen for the negative controls.

The Panel concluded that food enzyme β‐glucosidase did not induce chromosomal aberrations under the test conditions employed for this study.

Repeated dose 90‐day oral toxicity study in rodents

3.4.2

The repeated dose 90‐day oral toxicity study was performed in accordance with guidelines of the Japanese Ministry of Health and Welfare (1996 and 1999) and following GLP.19 The study is in accordance with OECD Test Guideline 408 (OECD, 1998) with the following deviations: detailed clinical observations and functional observations were not performed, urea was not determined, epididymides were not weighed, and only one area of the brain and one level of the spinal cord were examined in the microscopy. The Panel considered that these deviations are not major and do not impact on the evaluation of the study.

Groups of 12 male and 12 female Sprague–Dawley (Crj:CD(SD)IGS) rats received by gavage the food enzyme in doses of 100, 300 or 1000 mg/kg bw per day corresponding to 94.3, 282.9 or 943 mg TOS/kg bw per day. Controls received the vehicle (water for injection).

One high‐dose female died on day 85 due to malignant lymphoma, a neoplasm occasionally observed in SD rats. The Panel considered this death as being incidental.

The haematological investigation revealed a statistically significant prolonged prothrombin time in low‐dose males (+13%). The Panel considered this change as not toxicologically relevant as it was observed only in one sex and treatment group, and there were no changes in other relevant parameters (activated partial thromboplastin time, fibrinogen).

The clinical chemistry investigation revealed statistically significantly increased activity of alanine aminotransferase (ALT, +25%) and aspartate aminotransferase (AST, +39%) in low‐dose males, decreased total cholesterol (−19%), phospholipid (−19%) and calcium (−3%) in low‐dose males and increased potassium (+6%) in high‐dose males. The Panel considered the changes as not toxicological relevant as they were only observed in one sex (all parameters), there was no dose–response relationship (all parameters except for potassium) and the changes were small (ALT, AST).

A statistically significant decreased absolute brain weight (−4%) was reported in high‐dose males. The Panel considered the change as not toxicological relevant as the change was small, it was only observed in one sex and there were no histopathological changes in the brain.

No other statistically significant or biologically relevant differences to controls were observed.

The Panel identified the no observed adverse effect level (NOAEL) of 943 mg TOS/kg bw per day, the highest dose tested.

Allergenicity

3.4.3

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

The potential allergenicity of the β‐glucosidase produced with the P. guanacastense strain AE‐GLY was assessed by comparing its amino acid sequence with those of known allergens according to the ‘Scientific opinion on the assessment of allergenicity of GM plants and microorganisms and derived food and feed of the Scientific Panel on Genetically Modified Organisms’ (EFSA GMO Panel, 2010). Using higher than 35% identity in a sliding window of 80 amino acids as the criterion, no match was found.20

No information is available on oral and respiratory sensitisation or elicitation reactions of this β‐glucosidase. In addition, no allergic reactions after oral exposure to β‐glucosidases have been reported in the literature.21

Penicillium species are known to cause respiratory allergy (Shen et al., 2000). However, several studies have shown that individuals respiratorily sensitised can ingest corresponding allergens without acquiring clinical symptoms of food allergy (Armentia et al., 2009; Cullinan et al., 1997; Poulsen, 2004).

■■■■■, a known source of allergens, is present in the medium fed to the microorganisms. However, during the fermentation process, this product will be degraded and utilised by the microorganisms for cell growth, cell maintenance and production of enzyme protein. In addition, the microbial biomass and fermentation solids are removed. Taking into account the fermentation process and downstream processing, the Panel considered that potentially allergenic residues from this source were not expected to be present.

The Panel considered that the risk of allergic reactions upon dietary exposure to this food enzyme cannot be excluded, but the likelihood is low.

Dietary exposure

3.5

Intended use of the food enzyme

3.5.1

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

TABLE 2: Intended uses and use levels of the food enzyme as provided by the applicant. 22

The food enzyme is added to citrus fruit to obtain a citrus extract,25 which is then used to reconstitute juices. The food enzyme‐TOS remain in the citrus extract.

In wine processing, the food enzyme is added to grape must during maceration and fermentation26 to release volatile aglycones, improving the flavour of the wine. The food enzyme‐TOS remain in the wine.

In tea processing, the food enzyme is added to crude tea extract.27 The β‐glucosidase hydrolyses glycosides to release volatile aglycones. The food enzyme‐TOS remain in the tea extract.

In the production of plant‐based analogues of milk and milk products, the food enzyme is added to soy drinks.28 The β‐glucosidase hydrolyses isoflavone glucosides. The hydrolysis reduces off‐flavour of soy drinks for direct consumption or when used to make other foods. The food enzyme‐TOS remain in these soy products.

Based on data provided on thermostability (see Section 3.3.1), it is expected that the β‐glucosidase is inactivated in the food manufacturing processes listed in Table 2, except for juice and wine production, in which it may remain active, depending on the processing conditions.

Dietary exposure estimation

3.5.2

Chronic exposure to the food enzyme‐TOS was calculated 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 4.054 mg TOS/kg bw per day in children at the 95th percentile.

Uncertainty analysis

3.5.3

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 the exposure estimate to 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 (943 mg TOS/kg bw per day) from the 90‐day rat study with the derived exposure estimates of 0.008–2.3 mg TOS/kg bw per day at the mean and from 0.029 to 4.054 mg TOS/kg bw per day at the 95th percentile, resulted in a margin of exposure of at least 233.

CONCLUSIONS

4

Based on the data provided and the derived margin of exposure, the Panel concluded that the food enzyme β‐glucosidase produced with the non‐genetically modified Penicillium guanacastense strain AE‐GLY does not give rise to safety concerns under the intended conditions of use.

DOCUMENTATION AS PROVIDED TO EFSA

5

Application for authorisation of beta‐glucosidase from Penicillium multicolor AE‐GLY. January 2021. Submitted by Amano Enzyme Inc.

Additional information. February 2022. Submitted by Amano Enzyme Inc.

Abbreviationsbwbody weightCASChemical Abstracts ServiceCEPEFSA Panel on Food Contact Materials, Enzymes and Processing AidsCFUcolony forming unitsEINECSEuropean Inventory of Existing Commercial Chemical SubstancesFAOFood and Agricultural Organization of the United NationsGLPGood Laboratory PracticeGMMgenetically modified microorganismGMOgenetically modified organismIUBMBInternational Union of Biochemistry and Molecular BiologyJECFAJoint FAO/WHO Expert Committee on Food AdditiveskDakiloDaltonLoQlimit of quantificationMoEmargin of exposureOECDOrganisation for Economic Cooperation and DevelopmentTOStotal organic solidsWHOWorld Health Organization

CONFLICT OF INTEREST

If you wish to access the declaration of interests of any expert contributing to an EFSA scientific assessment, please contact interestmanagement@efsa.europa.eu.

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2015‐00273

COPYRIGHT FOR NON‐EFSA CONTENT

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PANEL MEMBERS

José Manuel Barat Baviera, Claudia Bolognesi, Andrew Chesson, Pier Sandro Cocconcelli, Riccardo Crebelli, David Michael Gott, Konrad Grob, Claude Lambré, Evgenia Lampi, Marcel Mengelers, Alicja Mortensen, Gilles Rivière, Inger‐Lise Steffensen, Christina Tlustos, Henk Van Loveren, Laurence Vernis, Holger Zorn.

NOTE

The full opinion will be published in accordance with Article 12 of Regulation (EC) No 1331/2008 once the decision on confidentiality will be received from the European Commission.

Supporting information

Dietary exposure estimates to the food enzyme–TOS in details