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Publication Abstract from PubMed

Mannose 2-epimerase (ME), a member of the acylglucosamine 2-epimerase (AGE) superfamily that catalyzes epimerization of D-mannose and D-glucose, has recently been characterized to have potential for D-mannose production. However, the substrate-recognition and catalytic mechanism of ME remains unknown. In this study, structures of Runella slithyformis ME (RsME) and its D254A mutant [RsME(D254A)] were determined in their apo forms and as intermediate-analog complexes [RsME-D-glucitol and RsME(D254A)-D-glucitol]. RsME possesses the (alpha/alpha)(6)-barrel of the AGE superfamily members but has a unique pocket-covering long loop (loop(alpha7-alpha8)). The RsME-D-glucitol structure showed that loop(alpha7-alpha8) moves towards D-glucitol and closes the active pocket. Trp251 and Asp254 in loop(alpha7-alpha8) are only conserved in MEs and interact with D-glucitol. Kinetic analyses of the mutants confirmed the importance of these residues for RsME activity. Moreover, the structures of RsME(D254A) and RsME(D254A)-D-glucitol revealed that Asp254 is vital for binding the ligand in a correct conformation and for active-pocket closure. Docking calculations and structural comparison with other 2-epimerases show that the longer loop(alpha7-alpha8) in RsME causes steric hindrance upon binding to disaccharides. A detailed substrate-recognition and catalytic mechanism for monosaccharide-specific epimerization in RsME has been proposed.

Structural insights into the substrate specificity and activity of a novel mannose 2-epimerase from Runella slithyformis.,Wang H, Sun X, Saburi W, Hashiguchi S, Yu J, Ose T, Mori H, Yao M Acta Crystallogr D Struct Biol. 2023 Jul 1;79(Pt 7):585-595. doi: , 10.1107/S205979832300390X. Epub 2023 Jun 14. PMID:37314406^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.