Yunqi Zhu, Fan Xu, Li Yang, Wenli Zhang, Y.-L. Chen, Wanmeng Mu
As essential prebiotics in breast milk, human milk oligosaccharides (HMOs) are critical for infant health including guiding the development of the gut microbiota, fine-tuning immune responses, and strengthening the intestinal barrier. These functions support the potential of HMOs to confer health benefits in adults as well. 2′-Fucosyllactose (2′-FL), a major trisaccharide in human milk, is a well-tolerated infant nutrient that is approved as a novel food in many countries. To date, 2′-FL can be efficiently synthesized by various cell factories via de novo and salvage pathways. Key strategies for increasing the 2′-FL titer include enhancing the GDP- l -fucose supply, introducing highly active α1,2-fucosyltransferases, and using diverse carbon sources to expand production flexibility. This review explores the health benefits of 2′-FL, along with key advances in the engineering of microbial cell factories and α1,2-fucosyltransferases for its production, concluding with a forward-looking perspective on priority research areas for enhancing its application.