Assessment of the feed additive consisting of l‐valine produced by fermentation with Corynebacterium glutamicum KCCM 80058 for all animal species for the renewal of its authorisation (CJ Europe GmbH)

TL;DR

This paper assesses the safety and authorization renewal of l-valine produced by a genetically modified bacterium for use in animal feed.

Contribution

The study confirms the continued safety of l-valine for animals, consumers, and the environment without requiring re-evaluation of its efficacy.

Findings

The use of l-valine in animal nutrition remains safe for target species, consumers, and the environment.

l-Valine produced by C. glutamicum KCCM 80058 is not an irritant to skin or eyes nor a skin sensitiser.

No modification proposal in the application affects the additive's efficacy, so re-assessment is unnecessary.

Abstract

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the assessment of the application for the renewal of the authorisation of l‐valine produced by fermentation with a genetically modified strain of Corynebacterium glutamicum KCCM 80058 as a nutritional feed additive for all animal species. The additive is authorised for use in all animal species (3c370). The applicant has provided evidence that the additive currently in the market complies with the existing conditions of authorisation. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the use of the feed additive in animal nutrition remains safe for the target species, consumers and the environment. As regards the safety for the user, l‐valine produced by C. glutamicum KCCM 80058 is not an irritant to skin or eyes nor a skin sensitiser. The present application for renewal of the authorisation does not include any modification proposal that would have an impact on the efficacy of the additive and therefore there is no need for re‐assessing the efficacy.

Full text

INTRODUCTION

1

Background and Terms of Reference

1.1

Regulation (EC) No 1831/20031 establishes the rules governing the Community authorisation of additives for use in animal nutrition. In particular, Article 14(1) of that Regulation lays down that an application for renewal shall be sent to the Commission at the latest one year before the expiry date of the authorisation.

The European Commission received a request from CJ Europe GmbH2 for the renewal of the authorisation of the additive consisting of L‐valine produced by fermentation with Corynebacterium glutamicum KCCM 80058, when used as a feed additive for all animal species (category: nutritional additives; functional group: amino acids, their salts and analogues).

According to Article 7(1) of Regulation (EC) No 1831/2003, the Commission forwarded the application to the European Food Safety Authority (EFSA) as an application under Article 14(1) (renewal of the authorisation). The dossier was received on 17 August 2023 and the general information and supporting documentation are available at https://open.efsa.europa.eu/questions/EFSA‐Q‐2023‐00551. The particulars and documents in support of the application were considered valid by EFSA as of 24 November 2023.

According to Article 8 of Regulation (EC) No 1831/2003, EFSA, after verifying the particulars and documents submitted by the applicant, shall undertake an assessment in order to determine whether the feed additive complies with the conditions laid down in Article 5. EFSA shall deliver an opinion on the safety for the target animals, consumer, user and the environment, and on the efficacy of the feed additive consisting of L‐valine produced by fermentation with a genetically modified strain of C. glutamicum (KCCM 80058), when used under the proposed conditions of use (see Section 3.1.3).

Additional information

1.2

The additive l‐valine produced by fermentation with C. glutamicum KCCM 80058 is currently authorised for use in feed for al animal species (3c370).3 EFSA issued an opinion on the safety and efficacy of this product when used in feed or water for drinking for all animal species (EFSA FEEDAP Panel).

DATA AND METHODOLOGIES

2

Data

2.1

The present assessment is based on data submitted by the applicant in the form of a technical dossier4 in support of the authorisation request for the use of l‐valine produced by fermentation with C. glutamicum KCCM 80058 as a feed additive.

In accordance with Article 38 of the Regulation (EC) No 178/20025 and taking into account the protection of confidential information and of personal data in accordance with Articles 39 to 39e of the same Regulation, and of the Decision of EFSA's Executive Director laying down practical arrangements concerning transparency and confidentiality,6 a non‐confidential version of the dossier has been published on Open.EFSA.

According to Article 32c(2) of Regulation (EC) No 178/2002 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 10 May to 31 May 2024, for which no comments were received.

The confidential version of the technical dossier was subject to a target consultation of the interested Member States from 24 November 2023 to 24 February 2024; the comments received were considered for the assessment.

The FEEDAP Panel used the data provided by the applicant together with data from other sources, such as previous risk assessments by EFSA or other expert bodies, peer‐reviewed scientific papers, other scientific reports and experts' knowledge, to deliver the present output.

The European Union Reference Laboratory (EURL) considered that the conclusions and recommendations reached in the previous assessment regarding the methods used for the control of the l‐valine produced by fermentation with C. glutamicum KCCM 80058 in animal feed are valid and applicable for the current application.7

Methodologies

2.2

The approach followed by the FEEDAP Panel to assess the safety and the efficacy of l‐valine produced by fermentation with C. glutamicum KCCM 80058 is in line with the principles laid down in Regulation (EC) No 429/20088 and the relevant guidance documents: Guidance on the renewal of the authorisation of feed additives (EFSA FEEDAP Panel, 2021).

ASSESSMENT

3

The product consisting of l‐valine produced by fermentation with a genetically modified strain of C. glutamicum (KCCM 80058) is currently authorised as a nutritional additive (functional group: amino acids, their salts and analogues) for all animal species. This assessment regards the renewal of the authorisation.

Characterisation

3.1

Characterisation of the production strain

3.1.1

l‐Valine is produced by a genetically modified strain of C. glutamicum, which is deposited at the Korean Culture Collection of Microorganisms (KCCM) with the deposition number of KCCM 80058.9 The strain was characterised in full in the first assessment by the FEEDAP Panel (EFSA FEEDAP Panel, 2013). The applicant has provided new data on the characterisation of the strain that are described below.

The taxonomic identification of the production strain, KCCM 80058, was confirmed ■■■■■ of the whole genome sequence (WGS) of the production strain, ■■■■■.10

The susceptibility of the production strain to relevant antibiotics was tested against the list of antimicrobials described for ‘Corynebacterium and other Gram‐positive’ in the Guidance on the characterisation of microorganisms used as feed additives or as production organisms (EFSA FEEDAP Panel, 2018).11 All measured minimum inhibitory concentration (MIC) values were equal or fell below the cut off values set in this Guidance and, therefore, the strain is considered susceptible to the relevant antibiotics.

The WGS data of the production strain were searched for the presence of antimicrobial resistance (AMR) genes ■■■■■12 ■■■■■13 ■■■■■ it can be concluded that overall, no genes of concern were identified.

The WGS data of the production strain were screened for the presence of genes coding for virulence factors ■■■■■. No genes of concern were identified.14

Information related to the genetically modified microorganism

15

3.1.1.1

Characterisation of the recipient or parental microorganism

■■■■■■■■■■

Description of the genetic modification

■■■■■17

Characterisation of the additive

3.1.2

l‐Valine (International Union of Pure and Applied Chemistry (IUPAC)) name: (2S)‐2‐amino‐3‐methylbutanoic acid; synonyms: α‐amino isovaleric acid, 2‐amino‐3‐methylbutyric acid, a compound identified by Chemical Abstracts Service (CAS) No 72‐18‐4 and European Inventory of Existing Commercial Chemical Substances (EINECS) No 200‐773‐6, has a molecular weight of 117.15 g/mol; the molecular formula is C_5_H_11_NO_2_.

The additive is currently authorised with a minimum content of l‐valine ≥ 98.0% on dry matter (DM) basis. The applicant declared that no modifications to the manufacturing process were performed since the previous application.18

Analytical data to confirm the specifications were provided for eight batches of the additive, showing the following average values: 98.7% DM (range 98.6%–98.8%). Loss on drying was on average 0.07% (range 0.02%–0.11%).19 The amount of unidentified material is up to 1.32%. Three additional batches of the additive were also analysed with another analytical method showing values of l‐valine ranging 98.9%–99.1% and moisture < 0.2%.20

Three batches of the additive were analysed for impurities.21 Cadmium, lead, mercury and arsenic concentrations were below the limit of quantification (LOQ) of the analytical methods.22

Polychlorinated dibenzo‐p‐dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin‐like polychlorinated biphenyls (DL‐PCBs) were analysed in three batches. All values were below the corresponding LOQ. The calculated upper bound (UB) concentration was 0.057 ng WHO (2005)‐TEQ/kg for the sum of PCDD/Fs, and 0.116 ng Van den Berg et al. (2006)‐TEQ/kg for the sum of PCDD/Fs and DL‐PCBs. The UB for the sum of non DL‐PCBs was 0.53 μg/kg (all values are expressed based on 88% DM).23

The analysis of mycotoxins, including aflatoxins (B1, G1, B2, G2), fumonisins (B1, B2, B3), ochratoxin A, deoxynivalenol, zearalenone, and T‐2 and HT‐2 toxins showed values below the LOQ.24

Microbiological contamination was analysed by determination of yeasts, moulds, Enterobacteriaceae, Escherichia coli and Salmonella spp. The results showed values of < 10 CFU/g for yeasts and moulds, no detection in 10 g for Enterobacteriaceae and E. coli, and no detection in 25 g for Salmonella spp.

The FEEDAP Panel considers that the amounts of the detected impurities do not raise safety concerns.

The presence of viable cells of the production strain was investigated in three batches of the final product, ■■■■■25 ■■■■■ No viable cells were detected.

The presence of DNA of the production strain in the final product was investigated in three batches of the final product ■■■■■26 ■■■■■ No DNA of the production strain was detected.

No new data have been provided regarding the physico‐chemical properties or stability of the additive. Since no changes were introduced in the manufacturing process, the data described in the previous opinion still apply (EFSA FEEDAP Panel, 2013).

Conditions of use

3.1.3

The additive is currently authorised for use in feed for all animal species without minimum or maximum content. Under ‘Other provisions’ it is stated:

1. Declarations to be made on the labelling of the additive:

• –moisture content.

2. Where voluntary declaration of the additive is made on the labelling of feed materials and compound feed, the following shall be included:

• –name and identification number of the additive,
• –added amount of the additive.

The applicant did not request any change in the current conditions of authorisation.

Safety

3.2

The applicant declares that no adverse effects including accidents for target animals, consumers, users and/or the environment have been reported since the approval of the additive.27

Safety of the production organism

3.2.1

The production strain was identified as a C. glutamicum that qualifies for the qualified presumption of safety (QPS) approach to safety assessment when used for production purposes (EFSA BIOHAZ Panel, 2023b), does not carry acquired antimicrobial resistance genes, the genetic modifications performed are considered safe, and it is susceptible to antimicrobials of clinical human and veterinary relevance. No viable cells or DNA of the production strain were found in the final product. Therefore, the final product does not give raise to any safety concern regarding the production strain.

Safety for the target species, consumers and the environment

3.2.2

In its previous opinion, the FEEDAP Panel concluded that ‘l‐valine produced by C. glutamicum (KCCM 80058) is safe for all animal species. However, excess doses would create amino acid imbalances with negative consequences for animal performance. Correct dosing in formulating diets requires knowledge of the amino acid content in feed materials and of the requirements of animals. These data are all freely accessible. The FEEDAP Panel has concerns over the safety of l‐valine for target species when administered via water for drinking since any additional supply of an essential amino acid would disturb the balanced amino acid pattern of complete diets’, ‘The use of l‐valine produced by C. glutamicum (KCCM 80058) in animal nutrition is of no safety concern for the consumer’ and ‘The use of l‐valine in animal nutrition would not lead to any localised increase in its concentration in the environment. Therefore, the use of the product under application as a feed additive does not represent a risk to the environment’.

The FEEDAP Panel is not aware of any information that would modify its previous conclusions. Therefore, the Panel concludes that the additive l‐valine produced with C. glutamicum (KCCM 80058) remains safe for the target species, consumers and the environment under the approved conditions of the authorisation.

Safety for the user

3.2.3

In the previous assessment (EFSA FEEDAP Panel, 2013), the FEEDAP Panel concluded that ‘l‐valine produced by C. glutamicum (KCCM 80058) has been shown not to be an irritant or a dermal sensitiser. The results of an acute inhalation study performed at a high concentration show no evidence of significant acute toxicity by this route. Exposure of users by inhalation cannot be excluded’.

No new data have been submitted for the renewal of the authorisation regarding the effects of the additive under assessment. Therefore, the FEEDAP Panel reiterates the previous conclusions.

Efficacy

3.3

The present application for renewal of the authorisation does not include a proposal for amending or supplementing the conditions of the original authorisation that would have an impact on the efficacy of the additive. Therefore, there is no need for assessing the efficacy of the additive in the context of the renewal of the authorisation.

Post‐market monitoring

3.4

The FEEDAP Panel considers that there is no need for specific requirements for a post‐market monitoring plan other than those established in the Feed Hygiene Regulation28 and Good Manufacturing Practice.

CONCLUSIONS

4

The applicant has provided evidence that the additive currently in the market complies with the existing conditions of authorisation.

The Panel considers that the use of l‐valine produced with C. glutamicum KCCM 80058 remains safe for the target animals, consumers and the environment under the authorised conditions of use. As regards the safety for the user, l‐valine produced with C. glutamicum KCCM 80058 is not an irritant to skin or eyes nor a skin sensitiser.

There is no need to assess the efficacy of the additive in the context of the renewal of the authorisation.

ABBREVIATIONSAMRantimicrobial resistanceANIaverage nucleotide identityCASChemical Abstracts ServiceCFUcolony forming unitDMdry matterEINECSEuropean Inventory of Existing Commercial Chemical SubstancesFEEDAPEFSA Panel on Additives and Products or Substances used in Animal FeedKCCMKorean Culture Collection of MicroorganismsLODlimit of detectionLOQlimit of quantificationMICminimum inhibitory concentrationQPSqualified presumption of safety■■■■■■■■■■WGSwhole genome sequence

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐2023‐00551

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PANEL MEMBERS

Roberto Edoardo Villa, Giovanna Azimonti, Eleftherios Bonos, Henrik Christensen, Mojca Durjava, Birgit Dusemund, Ronette Gehring, Boet Glandorf, Maryline Kouba, Marta López‐Alonso, Francesca Marcon, Carlo Nebbia, Alena Pechová, Miguel Prieto‐Maradona, Ilen Röhe, and Katerina Theodoridou.

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.