Safety of the extension of use of 3‐fucosyllactose (3‐FL) as a novel food pursuant to Regulation (EU) 2015/2283

TL;DR

This paper evaluates the safety of increasing the use of 3-fucosyllactose (3-FL) in various food products, concluding it is safe under the proposed conditions.

Contribution

The paper provides a safety assessment for extending the use of 3-FL in infant and medical foods, confirming its safety based on intake estimates.

Findings

The estimated daily intake of 3-FL from the proposed extension is lower than natural intake from breast milk in infants.

The combined exposure from authorized and proposed uses remains below the highest natural intake from breast milk.

The Panel concludes that the proposed extension of use does not affect the safety of 3-FL.

Abstract

Following a request from the European Commission, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on the safety of the extension of use of 3‐fucosyllactose (3‐FL) as a novel food (NF) pursuant to Regulation (EU) 2015/2283. The NF, produced with a genetically modified strain (Escherichia coli BL21 (DE3) JBT‐3FL) of E. coli BL21 (DE3), is already authorised as ingredient in several food categories, including infant formula (IF) and follow‐on formula (FOF). The applicant proposed to increase the maximum use levels of the NF in IF, FOF, food for special medical purposes (FSMP) and food supplements (FS). EFSA estimated the anticipated daily intake of the NF from the proposed extension of use in the relevant food categories. Additionally, a new intake estimate including the already authorised conditions of use in other food categories was carried out. The estimated daily intake of the NF from high consumption of IF alone at the proposed maximum use level in infants < 16 weeks of age is lower than the estimated natural highest mean daily intake of 3‐FL from human milk in breastfed infants. The estimated highest 95th percentile daily intake in infants from the combined exposure from the maximum authorised and proposed uses of 3‐FL, is somewhat higher than the estimated intake from the already authorised uses and below the estimated highest natural mean daily intake of 3‐FL from human milk in breastfed infants. The same applies to the use of the NF in FSMP and FS. Therefore, the Panel considers that the proposed extension of use does not affect the safety of the NF and concludes that the NF, 3‐FL, is safe under the proposed conditions of use.

Tables

TABLE 1. Estimated mean daily intakes (mg/kg bw per day) of 3‐FL from average (800 mL) and high (1200 mL) daily intakes of human milk for infants (from 0 to 6 months) of 6.7 kg bw, based on the mean of mean concentrations (0.91 g/L) and the maximum mean concentration (5.00 g/L) of 3‐FL in mature human milk (EFSA, 2024).

	Mean daily intake (mg/kg bw) from 800 mL/day of human milk		Mean daily intake (mg/kg bw) from 1200 mL/day of human milk
	Mean of mean concentrations	Maximum mean concentration	Mean of mean concentrations	Maximum mean concentration
3‐FL	109	597	163	896

TABLE 2. Intake estimate (mg/kg bw per day) resulting from the use of the NF as an ingredient in the intended food categories at the maximum proposed use levels.

Population group	Age (years)	Mean intake (mg/kg bw per day)		P95 intake (mg/kg bw per day)
		Lowest a	Highest a	Lowest b	Highest b
Infants	< 1	43	182	157	387
Young children c	1 to < 3	3	56	24	150
Other children	3 to < 10	0	3	0	19
Adolescents	10 to < 18	0	1	0	3
Adults d	≥ 18	0	0	0	1

TABLE 3. Use of the novel food (NF) in food supplement (FS) and resulting intake expressed as mg/kg bw per day.

Population group	Age (years)	Body weight a (kg)	Use level (g/d)	Intake (mg/kg bw per day) b
Other children	3 to < 10	23.1	4.0	173
Young adolescents	10 to < 14	43.4	4.0	92
Old adolescents	14 to < 18	61.3	4.0	65
Adults	≥ 18	70.0	4.0	57

TABLE 4. Combined uses of 3‐FL from both authorised and proposed food categories at the maximum use level and resulting intake expressed as mg/kg per day.

Population group	Age (years)	Mean intake (mg/kg bw per day)		P95 intake (mg/kg bw per day)
		Lowest a	Highest a	Lowest b	Highest b
Infants	< 1	72	223	182	433
Young children c	1 to < 3	56	131	145	278
Other children	3 to < 10	22	70	49	129
Adolescents	10 to < 18	5	27	18	58
Adults d	≥ 18	11	14	27	34

Full text

INTRODUCTION

1

Background and Terms of Reference as provided by the requestor

1.1

On 25 April 2024, the applicant Chr. Hansen A/S submitted a request to the European Commission (EC) in accordance with Article 10 of Regulation (EU) 2015/2283,1 to authorise changes in the conditions of use of the novel food (NF) 3‐fucosyllactose (3‐FL) produced with a genetically modified strain (Escherichia coli BL21 (DE3) JBT‐3FL) of E. coli BL21 (DE3), which was authorised by Commission Implementing Regulation (EU) 2023/52.2

The application requests to authorise the increase in the maximum authorised use levels of 3‐FL produced with a genetically modified strain (E. coli BL21 (DE3) JBT‐3FL) of E. coli BL21 (DE3) in infant formulae (IF), follow‐on formulae (FOF) and foods for special medical purposes (FSMP) for infants and young children as defined in Regulation (EU) 609/20133 from the current levels of 0.9 and 1.2 g/L, respectively, to 1.75 g/L for all three food categories.

The application also requests the increase in the maximum authorised use levels of this NF in food supplements (FS) as defined in Directive 2002/46/EC4 intended for the general population excluding infants and young children, from the currently authorised levels of 3.0 to 4.0 g/day.

In accordance with Article 10(3) of Regulation (EU) 2015/2283, the EC asks the European Food Safety Authority (EFSA) to provide a scientific opinion on the safety of the proposed changes in the conditions of use of the NF 3‐FL produced with a genetically modified strain (E. coli BL21 (DE3) JBT‐3FL) of E. coli BL21 (DE3).

Additional information

1.2

3‐FL is included in the Union list of authorised NFs (Commission Implementing Regulation (EU) 2017/24705 of 20 December 2017) when produced by fermentation with genetically modified strains of E. coli K‐12 MG1655 (Commission Implementing Regulation (EU) 2021/20296), E. coli K‐12 DH1 (Commission Implementing Regulation (EU) 2023/22107) and E. coli BL21 (DE3) (Commission Implementing Regulation (EU) 2023/52^2^).

DATA AND METHODOLOGIES

2

Data

2.1

The safety assessment of this NF is based on data supplied in the application.

Administrative and scientific requirements for NF applications referred to in Article 10 of Regulation (EU) 2015/2283 are listed in Commission Implementing Regulation (EU) 2017/2469.8

A common and structured format on the presentation of NF applications is described in the EFSA guidance on the preparation and presentation of a NF application (EFSA NDA Panel, 2021a). As indicated in this guidance, it is the duty of the applicant to provide all the available (proprietary, confidential and published) scientific data (including both data in favour and not in favour) that are pertinent to the safety of the NF.

The applicant has submitted a confidential and a non‐confidential version of a dossier following the ‘EFSA guidance on the preparation and presentation of a NF application’ (EFSA NDA Panel, 2021a) and the ‘Administrative guidance for the preparation of applications on novel foods pursuant to Article 10 of Regulation (EU) 2015/2283’ (EFSA, 2021).

In accordance with Art. 38 of Regulation (EC) No 178/20029 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,10 the non‐confidential version of the dossier has been published on Open.EFSA.11

According to Art. 32c(2) of Regulation (EC) No 178/2002^9^ 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 (PC‐1284) on the non‐confidential version of the technical dossier from 21 January to 11 February 2025 for which no comments were received.

This NF application does not include a request for the protection of proprietary data.

Methodologies

2.2

The assessment follows the methodology set out in the EFSA Guidance on NF applications (EFSA NDA Panel, 2021a) and the principles described in the relevant existing guidance documents from the EFSA Scientific Committee. The legal provisions for the assessment of novel foods are laid down in Article 11 of Regulation (EU) 2015/2283^1^ and in Article 7 of Commission Implementing Regulation (EU) 2017/2469.^8^

The legal provisions for the assessment of food intended for infants and young children and FSMP, are laid down in Regulation (EU) 609/2013^3^ and Commission Delegated Regulation (EU) 2016/12812 (FSMP), and in Commission Delegated Regulation (EU) 2016/12713 (as regards the specific compositional and information requirements for IF and FOF and as regards requirements on information relating to infant and young child feeding).

This assessment concerns only the risks that might be associated with consumption of the NF under the proposed conditions of use and is not an assessment of the efficacy of the NF with regard to any claimed benefit. This assessment also is not an assessment on whether the NF is suitable as stipulated by Regulation (EU) No 609/2013^3^.

ASSESSMENT

3

Introduction

3.1

The NF which is the subject of the application is 3‐FL, a human‐identical milk oligosaccharide (HiMO). The NF is already authorised (Regulation (EU) 2017/2470^5^) and no changes in the production process were introduced as per the previous safety assessment (EFSA NDA Panel, 2022a; Regulation (EU) 2023/52^2^). The applicant requests an extension of use of the NF by proposing an increase of the maximum use levels of the NF as an ingredient in IF and FOF from the authorised 0.9 and 1.2 g/L, respectively, to 1.75 g/L for both, and in FSMP from 0.9 to 1.75 g/L or kg. The target populations for the proposed extension of use are infants and young children. In addition, the applicant also requests an increased maximum use level in FS from 3 to 4 g/day for the general population, excluding infants and young children.

According to Regulation (EU) 2015/2283^1^, the NF falls under the following categories:

• ‘food with a new or intentionally modified molecular structure, where that structure was not used as, or in, a food within the Union before 15 May 1997’; and
• ‘food consisting of, isolated from or produced from microorganisms, fungi or algae’.

Identity of the NF

3.2

The NF is 3‐FL produced with a genetically modified strain (E. coli BL21 (DE3) JBT‐3FL) of E. coli BL21 (DE3).

The applicant stated that there is no change to the identity of 3‐FL as currently approved in the Union list^5^.

Production process

3.3

The applicant stated that the manufacturing conditions for the NF have not changed.

History of use of the NF and/or of its source

3.4

History of use of the NF

3.4.1

The NF is authorised as an ingredient in IF and FOF, and in milk‐based drinks and similar products intended for young children, processed cereal‐based foods for infants and young children (see Appendix A).

Specifically, in IF and FOF the NF is authorised up to 0.9 and 1.2 g/L, respectively, in the final product ready for use, marketed as such or reconstituted as instructed by the manufacturer. In FSMP it is authorised for infants and young children up to 0.9 g/L or kg (infants from 0 until 6 months) and 1.2 g/L or kg (infants of 6–12 months and young children).

Finally, the NF is also authorised in FS up to 3 g/day in the general population, excluding infants and young children.

Intake of 3‐FL from human milk

3.4.2

In previous EFSA opinions (EFSA NDA Panel, 2021b, 2022a, 2023), the daily intake of 3‐FL from the consumption of human milk was estimated for a 6.7 kg body weight (bw) infant (EFSA Scientific Committee, 2012), considering the average and high daily intakes of human milk (800 and 1200 mL, respectively) for infants from 0 to 6 months (EFSA NDA Panel, 2013). Average (1.24 g/L) and maximum (1.44 g/L) concentrations of 3‐FL in human milk reported by Thurl et al. (2017) were used in intake estimations (EFSA NDA Panel, 2021b). More recently, in consideration of the large and recent data set included in the review by Soyyılmaz et al. (2021), the Panel decided to use the values reported there for the intake assessments of HiMOs (e.g. EFSA NDA Panel, 2022a, 2023). For 3‐FL, those values correspond to the mean of mean concentrations (0.92 g/L) and the maximum mean concentration (2.57 g/L) across studies and were considered as representative of the concentration range found in mature human milk. Finally, new data reported in a recent scientific and technical assistance report (EFSA, 2024), further to an extensive literature search, suggested the use of higher concentration data in human milk (respectively, 0.91 and 5.00 g/L). The estimated natural intakes of 3‐FL reported in Table 1 are based on the newly reported data.

Proposed uses and use levels and anticipated intake

3.5

Target population

3.5.1

The target populations proposed by the applicant for the extension of use of the NF in IF, FOF and FSMP are infants and young children. For the use of the NF in FS, the target population is the general population with exclusion of infants and young children.

Proposed uses and use levels

3.5.2

The applicant proposes an increase in the authorised maximum levels of the NF in IF and FOF from the authorised 0.9 and 1.2 g/L, respectively, to 1.75 g/L for both and in FSMP from 0.9 to 1.75 g/L or kg. For the category FSMP, the Panel notes that the use should be in accordance with the particular nutritional requirements of the persons for whom the products are intended according to Regulation (EU) No 609/2013^3^.

Finally, the applicant proposes an increase in the authorised maximum levels of the NF in FS from 3 to 4 g/day for the general population with exclusion of infants and young children.

Anticipated intake of the NF

3.5.3

Anticipated intake of the NF from the consumption of the NF in IF for infants up to 16 weeks of age

IF is expected to be the only food consumed by infants aged 0–16 weeks who are not breastfed. A high consumption of IF has been estimated to be 260 mL/kg bw per day for infants aged 0–16 weeks (EFSA Scientific Committee, 2017). Based on the proposed maximum use level of the NF in IF (1.75 g/L), the highest daily intake of the NF from IF alone is estimated to be 455 mg/kg bw per day.

The Panel notes that the anticipated daily intake of the NF from high consumption of IF alone at the proposed maximum use level is lower than the estimated highest natural mean daily intake of 3‐FL from human milk of 896 mg/kg bw (see Table 1) and similar to the previously estimated natural intake of 460 mg/kg bw based on Soyyılmaz et al. (2021) (EFSA NDA Panel, 2022a).

Anticipated intake of the NF from the proposed uses and use levels

EFSA performed an intake assessment of the anticipated daily intake of the NF based on the applicant's proposed extension of use in IF and FOF (1.75 g/L), also including the already authorised conditions of use of the NF as ingredient in other food categories (as for EFSA NDA Panel, 2022a; Regulation (EU) 2023/52^2^ ‐ see Appendix A) and using the EFSA Dietary Exposure (DietEx) tool, which is based on individual data from the EFSA Comprehensive European Food Consumption Database (EFSA, 2011). The lowest and highest mean and 95th percentile anticipated daily intakes of the NF (on a kg bw basis), among the EU dietary surveys, are presented in Table 2.

The estimated daily intake of the NF for each population group from each EU dietary survey is available in the excel file annexed to this scientific opinion (under supporting information).

The Panel notes that the anticipated daily intake of the NF from the proposed uses and use levels does not exceed the estimated highest natural mean daily intake of 3‐FL from human milk of 896 mg/kg bw per day (Table 1).

Anticipated intake of the NF from the use in FS

The applicant has proposed a maximum daily intake of the NF of 4.0 g in FS for individuals above 3 years of age (Table 3).

The Panel notes that the maximum daily intake of the NF from its use in FS (i.e. from 57 to 173 mg/kg bw per day) for any population category (Table 3) does not exceed the estimated highest natural mean daily intake of 3‐FL from human milk of 896 mg/kg bw (Table 1). According to the applicant, FS containing the NF are not intended to be used if other foods with added 3‐FL are also consumed on the same day.

Combined intake from the NF and other sources

3.5.4

The Panel notes that 3‐FL produced with genetically modified strains of E. coli K‐12 MG1655 and E. coli K‐12 DH1 is already authorised for use in food categories other than those proposed for the NF under assessment (e.g. use in beverages, flavoured and unflavoured fermented milk‐based products, cereal bars).^5^

The combined daily intake of 3‐FL from the authorised and proposed uses, for each population group from each EU dietary survey, is available in the Excel file annexed to this scientific opinion (under supporting information) (Table 4).

The Panel notes that the estimated highest 95th percentile daily intake in infants from the combined exposure (i.e. 433 mg/kg bw) from the maximum authorised and proposed uses, is somewhat higher than the estimated intake from the already authorised uses (i.e. 366 mg/kg bw per day; EFSA NDA Panel, 2022a), and below the estimated highest natural mean daily intake of 3‐FL from human milk in infants (i.e. 896 mg/kg bw; Table 1).

Nutritional information

3.6

The NF consists of a non‐digestible oligosaccharide, 3‐FL. The highest P95 daily intake of the NF from the proposed extensions of use (including the authorised conditions of use in other food categories) is similar to that from the already authorised conditions of use of the NF as an ingredient.

The Panel considers that the consumption of the NF at the proposed use levels is not nutritionally disadvantageous.

Human data

3.7

No human intervention studies conducted with 3‐FL only have been provided by the applicant. However, the applicant made reference to two papers. In the first publication (Parschat et al., 2021) a multicentre, randomised, controlled, parallel‐group clinical study was conducted in infants with IF containing a mixture of HiMOs (composed by 3‐FL (13.0%), 2′‐fucosyllactose (2’‐FL) (52.0%), lacto‐N‐tetraose (LNT) (26.0%), 6′‐sialyllactose (6'‐SL) sodium salt (5.0%) and 3′‐sialyllactose (3’‐SL) sodium salt (4.0%)) at a concentration of 5.75 g/L, approximately corresponding to 0.75 g/L of 3‐FL. The safety and tolerability profile of the HiMO mixture–IF appeared similar to the IF used as control (containing the same quantities of proteins, lipids, vitamins and other nutrients) (EFSA NDA Panel, 2022a, 2022b, 2022c, 2023).

The second article (Lasekan et al., 2022) refers to another randomised, controlled, multicentre, double‐blind, parallel feeding growth and tolerance study with a mixture of 5 HiMOs. 3‐FL accounted for 0.8 g/L in the IF from the mixture. Other added HiMOs were: LNT, 3′‐SL, 6’‐SL and 2’‐FL, the latter being the most represented HiMO (3.0 g/L). All HiMOs were added according to average of the ranges and proportions of these five HMOs recorded in human milk. Three groups of healthy term infants were enrolled. One group was fed with an experimental formula containing the five HiMOs, the second group with a control milk‐based infant formula lacking HiMO and the third group was exclusively breastfed. According to the authors, the study demonstrated that the formula containing the HiMO mixture ‘supported normal growth, gastrointestinal tolerance and safe use in healthy term infants.’

The Panel considers the information provided by the applicant as supportive for the safety assessment of 3‐FL.

DISCUSSION

4

The NF which is the subject of the application is the already authorised HiMO 3‐FL. As specified in the Union list,^5^ the NF is produced by fermentation with a genetically modified strain (E. coli BL21 (DE3) JBT‐3FL) of E. coli BL21 (DE3). An increase in the maximum use levels of the NF in IF and FOF from the authorised 0.9 and 1.2 g/L, respectively, to 1.75 g/L, is proposed, while no changes in other authorised conditions of use of the NF are intended with exception of its use in FSMP (from 0.9 to 1.75 g/L or kg). IF is intended for use ‘by infants during the first months of life and satisfying by itself the nutritional requirements of such infants until the introduction of appropriate complementary feeding’ (Regulation (EU) No 609/2013^3^). Complementary foods are then necessary to meet infant's needs for energy and nutrients from the age of 4–6 months (EFSA NDA Panel, 2019; WHO, 2001, 2006). The Panel notes that the anticipated daily intake of the NF from high consumption of IF alone at the proposed maximum use level in infants up to 16 weeks of age (455 mg/kg bw) is lower than the newly estimated natural highest mean daily intake of 3‐FL from human milk in breastfed infants (896 mg/kg bw). The same applies for the other proposed use levels in other population groups and for the increased use level in FS from 3 to 4 g/day (in the general population with the exclusion of infants and young children (< 3 years)). In addition, FS containing the NF are not intended to be used if other foods (including human milk) containing 3‐FL are also consumed on the same day. Finally, the Panel also notes that the same use levels in IF and FOF are already authorised for 3‐FL produced by fermentation by genetically modified strains of E. coli K‐12 MG1655 and E. coli K‐12 DH1.

The Panel considers that the proposed increase of use levels of the NF in IF, FOF, FSMP and FS only marginally affects the highest P95 daily intake estimate from the authorised conditions of use of the NF as an ingredient and results in intakes that are lower than the estimated natural intake by breastfed infants on a body weight basis. Therefore, the Panel considers that this does not affect the safety of the NF.

CONCLUSIONS

5

The Panel concludes that the NF, 3‐FL, is safe under the proposed conditions of use.

STEPS TAKEN BY EFSA

6

1. On 21 June 2024 EFSA received a letter from the European Commission with the request for a scientific opinion on the safety of the use of of 3‐FL Ref.Ares(2024)4510000.
2. On 21 June 2024, a valid application on changes in the conditions of use of 3‐FL, which was submitted by Chr. Hansen A/S, was made available to EFSA by the European Commission e‐submission portal (NF 2024/25055) and the scientific evaluation procedure was initiated.
3. During its meeting on 24 March 2025, the NDA Panel, having evaluated the data, adopted a scientific opinion on the safety of the changes in the conditions of use of 3‐FL as a NF pursuant to Regulation (EU) 2015/2283.

ABBREVIATIONS2’‐FL2′‐fucosyllactose3‐FL3‐fucosyllactose3’‐SL3′‐sialyllactose6’‐SL6′‐sialyllactosebwbody weightDietExEFSA Dietary Exposure toolFOFfollow‐on formulaFSfood supplementFSMPfood for special medical purposesHiMOhuman‐identical milk oligosaccharideHMOhuman milk oligosaccharideIFinfant formulaLNTlacto‐N‐tetraoseNDA panelEFSA Panel on Nutrition, Novel Foods and Food AllergensNFnovel foodP9595th percentileWHOWorld Health Organization

REQUESTOR

European Commission

QUESTION NUMBER

EFSA‐Q‐ 2024‐00282

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PANEL MEMBERS

Dominique Turck, Torsten Bohn, Montaña Cámara, Jacqueline Castenmiller, Stefaan De Henauw, Karen Ildico Hirsch‐Ernst, Ángeles Jos, Alexandre Maciuk, Inge Mangelsdorf, Breige McNulty, Androniki Naska, Kristina Pentieva, Alfonso Siani, and Frank Thies.

Supporting information

Proposed uses and use levels of the NF

Combined uses from the NF and other sources