Safety evaluation of an extension of use of the food enzyme endo‐1,3(4)‐β‐glucanase from the non‐genetically modified Rasamsonia composticola strain 427‐FS

Abstract

The food enzyme endo‐1,3(4)‐β‐glucanase (3‐(1,3; 1,4)‐β‐D‐glucan 3(4)‐glucanohydrolase; EC 3.2.1.6) is produced with the non‐genetically modified Rasamsonia composticola strain 427‐FS by Kerry Ingredients & Flavours Ltd. The safety of this food enzyme was evaluated previously, and it did not give rise to safety concerns when used in six food manufacturing processes. Subsequently, the applicant has requested to extend its use to include two additional processes. In this assessment, EFSA updated the safety evaluation of this food enzyme when used in a total of eight food manufacturing processes. As the food enzyme–Total Organic Solids (TOS) are removed in two processes, the dietary exposure to the food enzyme–TOS was estimated for the remaining six food manufacturing processes. It was calculated to be up to 1.368 mg TOS/kg body weight (bw) per day in European populations. When combined with the no observed adverse effect level previously reported (866 mg TOS/kg bw per day, the highest dose tested), the Panel derived a margin of exposure of at least 633. Based on the revised exposure estimation, the newly derived margin of exposure and the previous evaluation, the Panel concluded that this food enzyme does not give rise to safety concerns under the revised 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 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.

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 places 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.

Endo‐1,3(4)‐β‐glucanase from non‐genetically modified Rasamsonia composticola (formerly Talaromyces emersonii) (strain 427‐FS) is a food enzyme included in the Register of food enzymes ³ to be considered for inclusion in the Union list and thus subject to a risk assessment by the European Food Safety Authority (EFSA).

On 29 November 2024, a new application was introduced by “Kerry Ingredients & Flavours Ltd.” Seeking authorization for an extension of the conditions of use of the food enzyme endo‐1,3(4)‐β‐glucanase from non‐genetically modified Rasamsonia composticola (strain 427‐FS).

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: extension of the conditions of use of the food enzyme endo‐1,3(4)‐ β‐glucanase from non‐genetically modified Rasamsonia composticola (strain 427‐FS) in accordance with Regulation (EC) No 1331/2008 establishing a common authorization procedure for food additives, food enzymes and food flavourings. ⁴

2. DATA AND METHODOLOGIES

2.1. Data

The applicant has submitted a dossier in support of the application for the authorisation of the extension of use of the food enzyme endo‐1,3(4)‐β‐glucanase from a non‐genetically modified Rasamsonia composticola strain 427‐FS.

Additional information was requested during the risk assessment phase. See https://open.efsa.europa.eu/questions/EFSA‐Q‐2025‐00534.

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, 2009) and following the relevant existing 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, 2023a) have been followed for the evaluation.

2.3. Public consultation

According to Article 32c(2) of Regulation (EC) No 178/2002 ⁵ 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 16 December 2025 to 6 January 2026. ⁶ No comments were received.

3. ASSESSMENT

IUBMB nomenclature	Endo‐1,3(4)‐β‐glucanase
Systematic name	3‐(1,3;1,4)‐β‐D‐Glucan 3(4)‐glucanohydrolase
Synonyms	endo‐1,3‐β‐D‐glucanase; laminarinase; β‐1,3‐glucanase
IUBMB No	3.2.1.6
CAS No	62213‐14‐3
EINECS No	263‐462‐4

Endo‐1,3(4)‐β‐glucanases catalyse the hydrolysis of 1,3‐ and 1,4‐β‐glycosidic linkages in mixed‐linked β‐D‐glucans, resulting in the generation of partially hydrolysed β‐D‐glucans.

All aspects concerning the safety of this food enzyme, when used in six food manufacturing processes, were evaluated in December 2022 (EFSA CEP Panel, 2023a).

Following a request to update the intended uses (adding two processes), EFSA revises the exposure assessment in the present evaluation and updates the safety evaluation of this food enzyme when used in eight food manufacturing processes.

3.1. Dietary exposure

The current dietary exposure supersedes section 3.5 of the previous evaluation (EFSA CEP Panel, 2023a).

3.1.1. Revised intended use of the food enzyme

The food enzyme is intended to be used in eight food manufacturing processes at the use levels summarised in Table 1.

TABLE 1.

Updated intended uses and use levels of the food enzyme. ⁷

Food manufacturing process	Raw material (RM)	Recommended use level (mg TOS/kg RM)
		Current evaluation a	Previous evaluation a , b
Processing of cereals and other grains
Production of baked products	Flour	17–50	17–50
Production of cereal‐based products other than baked	Flour	0.1–100	0.1–100
Production of brewed products	Cereals	0.6–50	0.6–50
Production of starch and gluten fractions	Cereals	4–75	4–75
Production of distilled alcohol	Cereals	5–20	5–20
Processing of plant‐ and fungal‐derived products
Production of plant extracts	Plant material	2–7
Production of plant‐based analogues of milk and milk products	Cereals, legumes, nuts, oil seeds	5–50
Processing of yeast and yeast products	Yeast biomass, autolysed yeast, yeast extract, cell walls	10–250	10–250

^a The numbers in bold were used for calculation.

^b The previous evaluation is made for the food enzyme application EFSA‐Q‐2021‐00746.

The two additional uses of the food enzyme are described below.

In production of plant extracts, the food enzyme is added to plant materials (e.g. aloe vera, stevia, fruit peelings, grains, etc.) ⁸ to decrease viscosity and to reduce processing time. The food enzyme–Total Organic Solids (TOS) remain in the final foods.

In production of plant‐based analogues of milk and milk products, the food enzyme is added to a slurry of plant‐based raw materials (e.g. cereals, legumes, oil seeds and nuts) ⁹ to decrease viscosity and to increase the yield. The food enzyme–TOS remain in the final foods.

The enzyme has a temperature optimum around 60°C (pH 5.0) and a pH optimum around 5.5 (50°C). After pre‐incubation of the food enzyme for 15 min at different temperatures, activity was unaffected by temperatures up to 65°C but was lost at 80°C (EFSA CEP Panel, 2023a). Based on these data and the downstream processing within the respective food manufacturing processes, the Panel concluded that the enzyme is inactivated in most of the food manufacturing processes listed in Table 1 in which the food enzyme–TOS remain. However, depending on the heat treatment conditions, it may remain in its active form in baked products, as a kinetic model, developed to predict enzyme inactivation during bread making, suggested that food enzymes may not be fully inactivated during baking (Zhang et al., 2017). Indeed, residual enzymatic activity was detected in white bread following the baking process (Reichenberger et al., 2020, 2025).

3.1.2. Dietary exposure estimation

In accordance with the guidance document (EFSA CEP Panel, 2021), dietary exposure was calculated for the six food manufacturing processes where the food enzyme–TOS remain in the final foods.

Chronic exposure to the food enzyme–TOS was calculated using the FEIM webtool ¹⁰ 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, 2023b).

Table 2 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 1.386 mg TOS/kg bw per day in infants at the 95th percentile.

TABLE 2.

Updated 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)	0.097–0.461 (14)	0.258–0.561 (17)	0.135–0.387 (21)	0.041–0.277 (23)	0.079–0.161 (23)	0.069–0.163 (25)
Min–max 95th percentile (number of surveys)	0.284–1.368 (13)	0.629–1.099 (16)	0.321–0.772 (21)	0.106–0.524 (22)	0.176–0.388 (23)	0.143–0.298 (24)

3.1.3. 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 3.

TABLE 3.

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	+/−
Exclusion of two processes from the exposure estimation: – production of distilled alcohol – production of starch and gluten fractions	−

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

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.

The exclusion of two food manufacturing processes from the exposure estimation was based on >99% of TOS removal. This is not expected to impact the overall estimate derived.

3.2. Margin of exposure

In the previous evaluation, the Panel identified a no observed adverse effect level (NOAEL) of 866 mg TOS/kg body weight (bw) per day, the highest dose tested, resulting in a margin of exposure (MoE) of at least 1070 (EFSA CEP Panel, 2023a).

A comparison of the NOAEL with the newly derived exposure estimates of 0.041–0.561 mg TOS/kg bw per day at the mean and from 0.106–1.368 mg TOS/kg bw per day at the 95th percentile resulted in an MoE of at least 633.

4. CONCLUSION

Based on revised exposure estimation, the newly derived margin of exposure and the previous evaluation, the Panel concluded that the food enzyme endo‐1,3(4)‐β‐glucanase produced with the non‐genetically modified Rasamsonia composticola strain 427‐FS does not give rise to safety concerns under the revised intended conditions of use.

5. DOCUMENTATION AS PROVIDED TO EFSA

Dossier and additional information can be accessed at https://open.efsa.europa.eu/dossier/FEN‐2024‐27194.

ABBREVIATIONS

bw

body weight

CAS

Chemical Abstracts Service

CEP

EFSA Panel on Food Contact Materials, Enzymes and Processing Aids

EC

European Commission

EINECS

European Inventory of Existing Commercial Chemical Substances

EU

European Union

IUBMB

International Union of Biochemistry and Molecular Biology

NOAEL

no observed adverse effect level

RM

raw material

TOS

Total Organic Solids

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2025‐00534

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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, Henk Van Loveren, Laurence Vernis and Holger Zorn.

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*, 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*, 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*, 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*, 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*, Netherlands, Poland, Portugal, Romania, Serbia*, Slovenia, Spain, Sweden

^*Consumption data from these pre‐accession countries are not reported in Table 2 of this opinion; however, they are included in Appendix A 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).

EFSA FEZ Panel (EFSA Panel on Food Enzymes) , Zorn, H. , Barat Baviera, J. M. , Bolognesi, C. , Catania, F. , Gadermaier, G. , Greiner, R. , Mayo, B. , Mortensen, A. , Roos, Y. H. , Solano, M. L. M. , Van Loveren, H. , Vernis, L. , Tokić, V. , Cavanna, D. , & Liu, Y. (2026). Safety evaluation of an extension of use of the food enzyme endo‐1,3(4)‐β‐glucanase from the non‐genetically modified Rasamsonia composticola strain 427‐FS . EFSA Journal, 24(7), e10215. 10.2903/j.efsa.2026.10215