化学
多糖
纤维素
荧光光谱法
色谱法
荧光团
氧化剂
溶解循环
荧光
低聚糖
单加氧酶
生物化学
酶
有机化学
生物
物理
量子力学
病毒学
细胞色素P450
病毒
作者
Thu V. Vuong,Bing Liu,Mats Sandgren,Emma R. Master
出处
期刊:Biomacromolecules
[American Chemical Society]
日期:2017-01-26
卷期号:18 (2): 610-616
被引量:36
标识
DOI:10.1021/acs.biomac.6b01790
摘要
Most existing methods for screening the activity of lytic polysaccharide mono-oxygenases (LPMOs) on polysaccharides are based on the detection of soluble oxidized sugars. This approach might underestimate the total performance of LPMOs since oxidation events that do not lead to oligosaccharide release are not detected. Using PcLPMO9D as a model enzyme, a microplate-based method has been developed to detect C1-oxidizing LPMO activity by covalently linking a water-soluble fluorophore to oxidized positions within the cellulose fiber. This fluorescence method was validated using X-ray photoelectron spectroscopy and then combined with high-performance anion-exchange chromatography to track total PcLPMO9D activity.
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