Structural insight into a CE15 esterase from the marine bacterial metagenome

• Published in: Scientific Reports
• Main Authors: De Santi, Concetta, Gani, Osman A. B. S. M., Helland, Ronny, Williamson, Adele Kim
• Format: Article in Journal/Newspaper
• Language: English
• Published: Nature Publishing Group 2017
• Subjects: Enzyme mechanisms; X-ray crystallography; VDP::Matematikk og Naturvitenskap: 400::Kjemi: 440; VDP::Mathematics and natural science: 400::Chemistry: 440; Arctic
• Online Access: https://hdl.handle.net/10037/12439; https://doi.org/10.1038/s41598-017-17677-4

Source at http://dx.doi.org/10.1038/s41598-017-17677-4 . The family 15 carbohydrate esterase (CE15) MZ0003, which derives from a marine Arctic metagenome, has a broader substrate scope than other members of this family. Here we report the crystal structure of MZ0003, which reveals that residues comprising the catalytic triad differ from previously-characterized fungal homologs, and resolves three large loop regions that are unique to this bacterial sub-clade. The catalytic triad of the bacterial CE15, which includes Asp 332 as its third member, closely resembles that of family 1 carbohydrate esterases (CE1), despite the overall lower structural similarity with members of this family. Two of the three loop regions form a subdomain that deepens the active site pocket and includes several basic residues that contribute to the high positive charge surrounding the active site. Docking simulations predict specific interactions with the sugar moiety of glucuronic-acid substrates, and with aromatically-substituted derivatives that serve as model compounds for the lignin-carbohydrate complex of plant cell walls. Molecular dynamics simulations indicate considerable flexibility of the sub-domain in the substrate-bound form, suggesting plasticity to accommodate different substrates is possible. The findings from this first reported structure of a bacterial member of the CE15 family provide insight into the basis of its broader substrate specificity.