Updated safety evaluation of the food enzyme glucan 1,4‐α‐maltohydrolase from the genetically modified Bacillus subtilis strain BABSC

TL;DR

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

Contribution

The study provides new data confirming the safety of a genetically modified Bacillus subtilis strain for food enzyme production.

Findings

The genetically modified Bacillus subtilis strain qualifies for the QPS approach.

No toxicological studies are needed for the enzyme under its intended use.

The enzyme is considered safe for food applications based on updated evaluations.

Abstract

The food enzyme glucan 1,4‐α‐maltohydrolase (4‐α‐d‐glucan α‐maltohydrolase; EC 3.2.1.133) is produced with the genetically modified Bacillus subtilis strain BABSC by Advance Enzyme Technologies, Ltd. In a previous opinion, the Panel on Food Contact Materials, Enzymes and Processing Aids of the European Food Safety Authority could not conclude on the safety of this food enzyme due to insufficient data to consider the production strain to qualify for the qualified presumption of safety (QPS) approach, and the absence of toxicological studies. The new data provided by the applicant showed that the production strain qualifies for the QPS approach and therefore, no toxicological studies are considered necessary. Based on the new data provided and the evaluation of the data previously submitted, the Panel concluded that this food enzyme does not give rise to safety concerns under the intended conditions of use.

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

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

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

• it does not pose a safety concern to the health of the consumer at the level of use proposed;
• there is a reasonable technological need;
• its use does not mislead the consumer.

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

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.

An application has been introduced by the applicant “Advanced Enzyme Technologies Limited” for the authorisation of the food enzyme Maltogenic amylase from a genetically modified Bacillus subtilis (strain BABSC).

The food enzyme was assessed by EFSA, which issued an inconclusive opinion on 5 December 2023. The new application contains supplementary information to address the data gaps identified in the scientific opinion (EFSA‐Q‐2017‐00546).

In accordance with Article 12(1) of Regulation (EC) No 234/2011 implementing Regulation (EC) No 1331/2008, the Commission has verified that the application falls within the scope of the food enzyme Regulation and contains all the elements required.

Terms of Reference

1.1.2

The Commission requests the European Food Safety Authority (EFSA) to verify the requirements of Article 32b of Regulation (EC) No 178/2002, the suitability of the data for risk assessment in accordance with Article 12(2) of Regulation (EU) No 234/2011, and to carry out the safety assessment and the assessment of possible confidentiality requests on the new data only for the following food enzyme: Maltogenic amylase from a genetically modified Bacillus subtilis (strain BABSC) 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.

Interpretation of the Terms of Reference

1.2

In December 2023, EFSA could not conclude on the safety evaluation of the food enzyme glucan 1,4‐α‐maltohydrolase from the genetically modified B. subtilis strain BABSC (EFSA CEP Panel, 2024). The present opinion is a follow‐up of the previous evaluation and assesses only supplementary information provided in the new dossier. Other aspects of the safety of the food enzyme were addressed in the previous Scientific Opinion.

DATA AND METHODOLOGIES

2

Data

2.1

The applicant has submitted a revised dossier in support of the application for authorisation of the food enzyme glucan 1,4‐α‐maltohydrolase from the genetically modified B. subtilis strain BABSC.

Additional information was requested during the risk assessment phase. See https://open.efsa.europa.eu/questions/EFSA‐Q‐2024‐00448.

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) has 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 6 to 27 December 2004, for which no comments were received.4

ASSESSMENT

3

IUBMB nomenclatureGlucan 1,4‐α‐maltohydrolaseSystematic name4‐α‐d‐glucan α‐maltohydrolaseSynonymsMaltogenase, Maltogenic α‐amylase, 1,4‐α‐d‐glucan α‐maltohydrolaseIUBMB NoEC 3.2.1.133CAS No160611‐47‐2EINECS No630‐523‐5

Glucan 1,4‐α‐maltohydrolases catalyse the hydrolysis of 1‐4‐α‐d‐glucosidic linkages in starch polysaccharides, releasing maltose units from the non‐reducing chain ends. The enzyme under assessment is intended to be used in baking processes and starch processing for the production of glucose syrups and other starch hydrolysates.

Source of the food enzyme

3.1

Previous data on the identification and characterisation of the production strain were insufficient to consider it to qualify for the qualified presumption of safety (QPS) approach. Based on this and in the absence of toxicological studies, the Panel was unable to conclude on the safety of the food enzyme glucan 1,4‐α‐maltohydrolase produced with the genetically modified B. subtilis strain BABSC under the intended conditions of use (EFSA CEP Panel, 2024).

The applicant has now provided new data for identification of the production strain. The whole genome sequence (WGS) of the production strain5 showed an average nucleotide identity (ANI) ■■■■■.6 Based on this, the production strain BABSC was identified as B. subtilis.

Furthermore, the applicant investigated the possible cytotoxicity of the production strain on VERO cells, using propidium iodide staining as a measure of cell disruption. The results showed that the production strain B. subtilis BABSC was not cytotoxic.7

In view of the above and since the genetic modifications did not raise safety concern (EFSA CEP Panel, 2024), all the qualifications have been met and the production strain is considered to qualify for the QPS approach.

Toxicological data

3.2

As the production strain qualifies for the QPS approach of safety assessment and no issue of concern arising from the production process of the food enzyme was identified (EFSA CEP Panel, 2024), the Panel considered that no toxicological studies are necessary.

CONCLUSIONS

4

Based on the new data, which shows the QPS status of the production strain, and the evaluation of the data previously submitted, the Panel concludes that the food enzyme glucan 1,4‐α‐maltohydrolase, produced with the genetically modified B. subtilis strain BABSC, does not give rise to safety concerns under the intended conditions of use.

DOCUMENTATION AS PROVIDED TO EFSA

5

Dossier and additional information can be accessed at: https://open.efsa.europa.eu/dossier/FEN‐2024‐26925.

ABBREVIATIONSANIaverage nucleotide identityCASChemical Abstracts ServiceCEPEFSA Panel on Food Contact Materials, Enzymes and Processing AidsEINECSEuropean Inventory of Existing Commercial Chemical SubstancesIUBMBInternational Union of Biochemistry and Molecular BiologyQPSqualified presumption of safetyWGSwhole genome sequence

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2024‐00448

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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, Holger Zorn.

LEGAL NOTICE

Relevant information or parts of this scientific output have been blackened in accordance with the confidentiality requests formulated by the applicant pending a decision thereon by EFSA. The full output has been shared with the European Commission, EU Member States (if applicable) and the applicant. The blackening may be subject to review once the decision on the confidentiality requests is adopted by EFSA and in case it rejects some of the confidentiality requests.