Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of L‐arginine produced with a genetically modified strain of Escherichia coli (CCTCC M 20231961) as a nutritional feed additive for all animal species and categories. The EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the production strain E. coli CCTCC M 20231961 does not raise safety concerns. No viable cells or DNA of the production strain are detected in the final product. Therefore, the FEEDAP Panel concludes that the additive does not pose any safety concern regarding the production strain. The use of L‐arginine produced by fermentation with E. coli CCTCC M 20231961 in feed is safe for the target species when supplemented in appropriate amounts to the diet according to the nutritional needs of the target species. The FEEDAP Panel has concerns on the use of L‐arginine in water for drinking. The use of L‐arginine produced by fermentation with E. coli CCTCC M 20231961 in animal nutrition is considered safe for the consumers and for the environment. The FEEDAP Panel cannot conclude on the potential of the additive to be irritant to skin and/or eyes and to be a potential dermal sensitiser. The feed additive consisting of L‐arginine produced by fermentation with E. coli CCTCC M 20231961 is regarded as an efficacious source of the amino acid L‐arginine for all non‐ruminant species. For the supplemental L‐arginine to be as efficacious in ruminants as in non‐ruminant species, it requires protection against degradation in the rumen.