Tangled up in fibers: How a lytic polysaccharide monooxygenase binds its chitin substrate

Abstract

Lytic polysaccharide monooxygenases (LPMOs) are redox-enzymes that bind to and oxidize insoluble carbohydrate substrates, such as chitin or cellulose. This class of enzymes has attracted considerable attention due to their ability to convert biomaterials of high abundance into oligosaccharides that can be useful for producing biofuels and bioplastics. However, processes at the interface between solution and insoluble substrates represent a major challenge to biochemical and structural characterization. This study used the four-domain LPMO from Vibrio cholerae, N-acetyl glucosamine binding protein A (GbpA), to elucidate how it docks onto its insoluble substrate with its two terminal domains. First, we developed a protocol that allowed GbpA and chitin to form a stable complex in suspension, overcoming incompatibilities of the two binding partners with respect to pH. After determining the neutron scattering contrast match point for chitin, we characterized the structure of GbpA in complex with chitin by Small-Angle Neutron Scattering (SANS). We found that GbpA binds rapidly to chitin, where it spreads out on the chitin fibers unevenly. These findings are supported by electron microscopy. Placing our findings into a biological context, we discussed the potential advantages of GbpA secretion and how chitin binding may prepare the ground for microcolony formation of the bacteria.

Calcium binding site in AA10 LPMO fromVibrio choleraesuggests modulating effects during environment survival and infection

“…GbpA is known to bind chitin through domains 1 and 4 13 , whereas binding to mucins appears to only involve the LPMO domain 1, 13 however, relatively little detail is known about these interactions at the molecular level. 30 An additional challenge is assessing what ionic compositions V. cholerae might be exposed to in its local microenvironment during environmental survival and intestinal colonization is a complex task.…”