Selective xyloglucan oligosaccharide hydrolysis by a GH31 alpha-xylosidase from Escherichia coli

Author(s)

Buffoni de Campos Carneiro, Lara Aparecida ; Fuzo, Carlos Alessandro ; Meleiro, Luana Parras ; Carli, Sibeli ; Barreto, Matheus Quintana ; Lourenzoni, Marcos Roberto ; Buckeridge, Marcos Silveira ; Ward, Richard John

• Document type: Journal article
• Source: Carbohydrate Polymers; v. 284, p. 12-pg., 2022-05-15.

Abstract

Xyloglucan is ubiquitous in the cell walls of land plants and is also an essential storage polymer in seeds of many species. We studied the hydrolysis of the non-reducing end xylosyl residue of xyloglucan oligosaccharides (XGOs) by the Escherichia coli alpha-xylosidase (YicI). Electrospray Ionization Tandem Mass Spectrometry (ESI-MS/MS) and ion fragmentation analysis together with high performance anion exchange chromatography with pulsed amperometric detection revealed that YicI preferentially removes the xylosyl residue from the glycosyl residue of non-galactosylated oligosaccharides. The YicI shows decreasing activity against the galactosylated oligosaccharides XXXG>XXLG >= XLXG. Studies of the XGOs interaction with active site residues by molecular dynamics simulations suggested that hydrogen bond interactions between the D49 and galactosylated oligosaccharides play an important role in enzyme-XGO interactions. This was confirmed by site-directed mutagenesis, where the D49A mutant affected catalytic efficiency against galactosylated XGOs. Our findings advance xyloglucan disassembly models and highlight the importance of YicI for biotechnology applications. (AU)

• FAPESP's process: 12/24147-8 - Rational design of multifunctional enzymes for depolimerization of lignocelulosic material
• Grantee: Lara Aparecida Buffoni de Campos Carneiro
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/13734-3 - Engineering of an endopolygalacturonase by insertion of different Carbohydrate Binding Modules (CBMs): chimerogenesis and proximity effects
• Grantee: Sibeli de Carli
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/24139-6 - Design and Characterization of Multifunctional Xylanolytic Enzymes
• Grantee: Richard John Ward
• Support Opportunities: Regular Research Grants
• FAPESP's process: 16/17582-0 - Thermostability modulation of a ß-glucosidase stimulated by glucose and xylose using site directed glycosylation
• Grantee: Luana Parras Meleiro Garcia
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/50884-5 - INCT 2014: National Institute of Science and Technology of Bioethanol
• Grantee: Marcos Silveira Buckeridge
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 16/15708-7 - Characterization of novel hydrolysis products from the synergistic hydrolysis of xyloglucan by a beta-galactosidase/alpha-xylosidase mixture
• Grantee: Lara Aparecida Buffoni de Campos Carneiro
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate