Fucosylated human milk oligosaccharides (FHMOs), mainly including 2′-fucosyllactose (2′-FL), 3-fucosyllactose (3-FL), and difucosyllactose (DFL), are the most abundant neutral human milk oligosaccharides in breast milk. FHMOs have multiple physiological functions, such as preventing pathogen adhesion, regulating the immune system, promoting probiotic growth, and promoting neurological development. The biosynthesis methods, including enzymatic and precision fermentation, have the advantages of easy operation, environmental friendliness, and mild reaction conditions, and have become a research focus and hotspot for the synthesis of FHMOs. Herein, the latest research progress in the biosynthesis of FHMOs by enzymatic and precision fermentation was systematically summarized. The contents focused on the microbial chassis cells used for FHMOs synthesis, FHMOs synthesis pathways and bypass inhibition, as well as the exploration and modification of fucosyltransferases. The microbial chassis cells used for FHMOs synthesis were introduced. Escherichia coli was the most commonly used chassis cell, followed by microorganisms such as Bacillus sp. and yeasts, and emerging chassis cells were also widely developed, such as Nicotiana benthamiana. The FHMOs synthesis pathway and bypass inhibition were analyzed. The overall idea was to enhance the gene expression level of the FHMOs synthesis pathway and weaken the consumption of carbon resources by bypass metabolism, while balancing the coordinated expression of genes in the synthesis pathway and carbon central metabolism pathway. Subsequently, the exploration and modification of fucosyltransferases were discussed. Various bioinformatics methods were used to discover efficient and highly specific fucosyltransferases, which were effective methods to enhance the biosynthesis of FHMOs. Rational/semi rational molecular modification was used to further improve the performance of fucosyltransferases, which could maximize the production and product singularity of FHMOs. Finally, the future trends of FHMOs biosynthesis were proposed.