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.