Structural and functional insights into Uly1040, an ulvan lyase from polysaccharide lyase family 40

Ulvan is a major structural polysaccharide in marine green algae, and its enzymatic degradation releases bioactive oligosaccharides with promising biotechnological potential. Ulvan lyases are key to this process, yet most characterized enzymes belong to only a few polysaccharide lyase families, leaving the PL40 family largely unexplored. Here, we identify and characterize Uly1040, a novel PL40 ulvan lyase from Alteromonas macleodii, revealing an unprecedented two-domain architecture and a distinct His/Tyr catalytic mechanism. Structural and biochemical analyses show that Mn2+, His487, and Asp358 cooperatively activate His485 as the catalytic base, while Tyr305 acts as the catalytic acid-representing a mechanistic innovation in ulvan cleavage. Bioinformatic and phylogenetic analyses indicate that the PL40 lyases are widespread in marine environment, and this catalytic strategy is conserved among PL40 enzymes. This work uncovers a previously unknown enzymatic paradigm for ulvan degradation, deepening our understanding of marine polysaccharide utilization and microbial carbon cycling.