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Mass spectrometric fragmentation patterns discriminate C1- and C4-oxidised cello-oligosaccharides from their non-oxidised and reduced forms.
Carbohydrate Polymers ( IF 11.2 ) Pub Date : 2020-01-26 , DOI: 10.1016/j.carbpol.2020.115917
Peicheng Sun 1 , Matthias Frommhagen 1 , Maloe Kleine Haar 1 , Gijs van Erven 1 , Edwin J Bakx 1 , Willem J H van Berkel 1 , Mirjam A Kabel 1
Affiliation  

Lytic polysaccharide monooxygenases (LPMOs) are powerful enzymes that degrade recalcitrant polysaccharides, such as cellulose. However, the identification of LPMO-generated C1- and/or C4-oxidised oligosaccharides is far from straightforward. In particular, their fragmentation patterns have not been well established when using mass spectrometry. Hence, we studied the fragmentation behaviours of non-, C1- and C4-oxidised cello-oligosaccharides, including their sodium borodeuteride-reduced forms, by using hydrophilic interaction chromatography and negative ion mode collision induced dissociation - mass spectrometry. Non-oxidised cello-oligosaccharides showed predominantly C- and A-type cleavages. In comparison, C4-oxidised ones underwent B-/Y- and X-cleavage close to the oxidised non-reducing end, while closer to the reducing end C-/Z- and A-fragmentation predominated. C1-oxidised cello-oligosaccharides showed extensively A-cleavage. Reduced oligosaccharides showed predominant glycosidic bond cleavage, both B-/Y- and C-/Z-, close to the non-reducing end. Our findings provide signature mass spectrometric fragmentation patterns to unambiguously elucidate the catalytic behaviour and classification of LPMOs.

中文翻译:

质谱碎裂模式将C1和C4氧化的纤维寡糖与它们的非氧化和还原形式区分开来。

溶解性多糖单加氧酶(LPMO)是可降解顽固性多糖(例如纤维素)的强大酶。但是,LPMO生成的C1和/或C4氧化的寡糖的鉴定远非简单易行。特别地,当使用质谱法时,它们的碎片化模式还没有很好地建立。因此,我们通过使用亲水相互作用色谱和负离子模式碰撞诱导解离-质谱法研究了非氧化,C1和C4氧化的纤维寡糖(包括其硼氢化氘还原形式)的断裂行为。未氧化的纤维寡糖主要显示C型和A型裂解。相比之下,C4氧化的分子在氧化的非还原端附近进行了B- / Y-和X裂解,而更靠近还原端的则是C- / Z-和A碎片。C1氧化纤维寡糖显示出广泛的A裂解。还原的寡糖在非还原端附近显示出主要的糖苷键裂解,B- / Y-和C- / Z-。我们的发现提供了标志性的质谱碎裂模式,以明确阐明LPMO的催化行为和分类。
更新日期:2020-01-26
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