Physiological effects, biosynthesis, and derivatization of key human milk tetrasaccharides, lacto-N-tetraose, and lacto-N-neotetraose

Human milk oligosaccharides (HMOs) have recently attracted ever-increasing interest because of their versatile physiological functions. In HMOs, two tetrasaccharides, lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT), constitute the essential components, each accounting 6% (w/w) of total HMOs. Also, they serve as core structures for fucosylation and sialylation, generating functional derivatives and elongation generating longer chains of core structures. LNT, LNnT, and their fucosylated and/or sialylated derivatives account for more than 30% (w/w) of total HMOs. For derivatization, LNT and LNnT can be modified into a series of complex fucosylated and/or sialylated HMOs by transferring fucose residues at α1,2-, α1,3-, and α1,3/4-linkage and/or sialic acid residues at α2,3- and α2,6-linkage. Such structural diversity allows these HMOs to possess great commercial value and an application potential in the food and pharmaceutical industries. In this review, we first elaborate the physiological functions of these tetrasaccharides and derivatives. Next, we extensively review recent developments in the biosynthesis of LNT, LNnT, and their derivatives in vitro and in vivo by employing advanced enzymatic reaction systems and metabolic engineering strategies. Finally, future perspectives in the synthesis of these HMOs using enzymatic and metabolic engineering approaches are presented.