Robust one-pot multi-enzyme polysaccharide remodeling strategy for the synthesis of uniform chondroitin fragments and derivatives.

Abstract The non-sulfated chondroitin backbone (CH) is the synthetic precursor of chondroitin sulfate, a linear polysaccharide with dramatic biological functions. Owing to the intrinsic characteristics of the polysaccharide biosynthetic pathway, it is still a challenge to obtain structural-defined glycans via microbial fermentation or enzymatic synthesis, which hindering the illustration of CH polysaccharide functions. Herein, we report a robust one-pot multi-enzyme polysaccharide remodeling strategy to synthesize uniform CH fragments and their derivatives. CH tetrasaccharide, which was obtained from the digestion of heterogeneous CH fragments, was used as the starting material to trigger the assembly of uniform CH fragments in a one-pot multi-enzyme system. This strategy, which combined heteropolymer digestion, sugar nucleotide in situ generation, and sugar chain synchronized polymerization, provides a robust toolbox for structural-defined polysaccharides synthesis.