Structural and Biochemical Analysis Reveals Catalytic Mechanism of Fucoidan Lyase from Flavobacterium sp. SA-0082

Fucoidans represent a type of polyanionic fucose-containing sulfated polysaccharides (FCSPs) that are cleaved by fucoidan-degrading enzymes, producing low-molecular-weight fucoidans with multiple biological activities suitable for pharmacological use. Most of the reported fucoidan-degrading enzymes are glycoside hydrolases, which have been well studied for their structures and catalytic mechanisms. Little is known, however, about the rarer fucoidan lyases, primarily due to the lack of structural information. FdlA from Flavobacterium sp. SA-0082 is an endo-type fucoidan-degrading enzyme that cleaves the sulfated fuco-glucuronomannan (SFGM) through a lytic mechanism. Here, we report nine crystal structures of the catalytic N-terminal domain of FdlA (FdlA-NTD), in both its wild type (WT) and mutant forms, at resolutions ranging from 1.30 to 2.25 angstrom. We show that the FdlA-NTD adopts a right-handed parallel beta-helix fold, and possesses a substrate binding site composed of a long groove and a unique alkaline pocket. Our structural, biochemical, and enzymological analyses strongly suggest that FdlA-NTD utilizes catalytic residues different from other beta-helix polysaccharide lyases, potentially representing a novel polysaccharide lyase family.

Automated summary

Fucoidans are polyanionic fucose-containing sulfated polysaccharides that can be cleaved by fucoidan-degrading enzymes to produce low-molecular-weight fucoidans suitable for pharmaceutical use. FdlA is an endo-type fucoidan-degrading enzyme that cleaves sulfated fuco-glucuronomannan through a lytic mechanism. The crystal structures of FdlA-NTD reveal a right-handed parallel beta-helix fold with unique substrate binding site and potentially representing a novel polysaccharide lyase family with different catalytic residues.