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Efficient separation of oxidized cello-oligosaccharides generated by cellulose degrading lytic polysaccharide monooxygenases

Westereng, Bjørge ; Agger, Jane Wittrup ; Horn, Svein J. ; Vaaje-Kolstad, Gustav ; Aachmann, Finn L. ; Stenstrøm, Yngve H. ; Eijsink, Vincent G.H.

Journal of Chromatography A, 2013-01, Vol.1271 (1), p.144-152 [Periódico revisado por pares]

Amsterdam: Elsevier B.V

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Título:
Efficient separation of oxidized cello-oligosaccharides generated by cellulose degrading lytic polysaccharide monooxygenases
Autor: Westereng, Bjørge ; Agger, Jane Wittrup ; Horn, Svein J. ; Vaaje-Kolstad, Gustav ; Aachmann, Finn L. ; Stenstrøm, Yngve H. ; Eijsink, Vincent G.H.
Assuntos: Alternative fuels. Production and utilization ; Analytical chemistry ; anion exchange chromatography ; Applied sciences ; Biofuel ; Biofuels ; biomass ; carbon ; cellobiose ; cellulose ; Cellulose - chemistry ; Cellulose - metabolism ; Cellulose aldonic acids ; Chemistry ; Chromatographic methods and physical methods associated with chromatography ; Chromatography, High Pressure Liquid - methods ; Chromatography, Ion Exchange - methods ; detectors ; Energy ; Exact sciences and technology ; Fuels ; glycosides ; high performance liquid chromatography ; High-performance anion-exchange chromatography (HPAEC) ; hydrolases ; hydrophilic interaction chromatography ; Hydrophilic interaction chromatography (HILIC) ; Hydrophobic and Hydrophilic Interactions ; light scattering ; Lytic polysaccharide monooxygenase (LPMO) ; mass spectrometry ; Miscellaneous ; Mixed Function Oxygenases - chemistry ; Mixed Function Oxygenases - metabolism ; nuclear magnetic resonance spectroscopy ; Oligosaccharides - chemistry ; Oligosaccharides - isolation & purification ; Oligosaccharides - metabolism ; Other chromatographic methods ; Oxidation-Reduction ; polymerization ; Porous graphitized carbon liquid chromatography (PGC-LC)
É parte de: Journal of Chromatography A, 2013-01, Vol.1271 (1), p.144-152
Notas: http://dx.doi.org/10.1016/j.chroma.2012.11.048
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Descrição: ► We show efficient separation of cello-oligosaccharides and their aldonic acids. ► Using HPAEC, run times can be as short as 10min. ► HILIC and PGC-LC methods are suitable for LC–MS set-ups. ► Cello-oligosaccharide aldonic acids were purified by preparative PGC-LC. ► These methods are useful when studying lytic polysaccharide monooxygenases. We present an evaluation of HPLC-based analytical tools for the simultaneous analysis of native and oxidized cello-oligosaccharides, which are products of enzymatic cellulose degradation. Whereas cello-oligosaccharides arise from cellulose depolymerization by glycoside hydrolases, oxidized cello-oligosaccharides are produced by cellobiose dehydrogenase and the recently identified copper dependent lytic polysaccharide monooxygenases (LPMOs) currently classified as CBM33 and GH61. The latter enzymes are wide-spread and expected to play crucial roles in further development of efficient enzyme technology for biomass conversion. Three HPLC approaches with well documented performance in the field of oligosaccharide analysis have been investigated: high-performance anion-exchange chromatography (HPAEC), hydrophilic interaction chromatography (HILIC) and porous graphitized carbon liquid chromatography (PGC-LC). HPAEC with pulsed amperometric detection (PAD) was superior for analysis of oxidized oligosaccharides, combining the best separation with superior sensitivity for oligosaccharide species with a degree of polymerization (DP) ranging from 1 to 10. Furthermore, the HPAEC method can be optimized for operation in a high-throughput manner (run time 10min). Both PGC-LC and HILIC allow reasonable run times (41 and 25min, respectively), with acceptable separation, but suffer from poor sensitivity compared to HPAEC-PAD. On the other hand, PGC-LC and HILIC benefit from being fully compatible with online mass spectrometry. Using an LC–MS setup, these methods will deliver much better sensitivity than what can be obtained with conventional detectors such as ultraviolet-, charged aerosol-, or evaporative light scattering and may reach sensitivities similar to or even better than what is obtained in HPAEC-PAD. Pure oxidized cello-oligosaccharide standards, ranging from DP2 to DP5, were obtained by semi-preparative PGC and characterized by MS and NMR analysis.
Editor: Amsterdam: Elsevier B.V
Idioma: Inglês

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Efficient separation of oxidized cello-oligosaccharides generated by cellulose degrading lytic polysaccharide monooxygenases

Westereng, Bjørge ; Agger, Jane Wittrup ; Horn, Svein J. ; Vaaje-Kolstad, Gustav ; Aachmann, Finn L. ; Stenstrøm, Yngve H. ; Eijsink, Vincent G.H.

Amsterdam: Elsevier B.V

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