Glycosylated cyclophellitol-derived activity-based probes and inhibitors for cellulases.
Glycosylated cyclophellitol-derived activity-based probes and inhibitors for cellulases.
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DOI：
10.1039/d0cb00045k
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发表时间：
2020-08-01
影响因子：
4.1
通讯作者：
Davies GJ
中科院分区：
文献类型：
--
作者：
de Boer C;McGregor NGS;Peterse E;Schröder SP;Florea BI;Jiang J;Reijngoud J;Ram AFJ;van Wezel GP;van der Marel GA;Codée JDC;Overkleeft HS;Davies GJ
纤维素酶和相关的β-1，4-葡聚糖酶是木质纤维素降解酶混合物的基本组分。β-1，4-葡聚糖酶活性的检测通常依赖于监测纯化的木质纤维素衍生的底物或合成的显色底物的分解，从而限制了可检测的活性，并使将活性追溯到复杂酶混合物内的特定组分变得复杂。作为快速检测和鉴定β-1，4-葡聚糖酶的工具，人们合成了一系列模拟β-1，4-葡聚糖低聚糖的糖基化环菲醇抑制剂。这些化合物是众所周知的GH 7内切-β-1，4-葡聚糖酶HiCel 7 B的高效抑制剂。一个精心设计的基于活性的探针有助于直接检测和鉴定复杂的真菌分泌组中的β-1，4-葡聚糖酶，而没有任何可检测的交叉反应性与β-d-葡糖苷酶。这些探针和抑制剂增加了宝贵的新的能力，不断增长的工具箱的cyclophellitol衍生的探针的生物质降解酶的活性为基础的分析。新的环岩藻醇衍生的基于活性的探针能够灵敏地检测和鉴定纤维素酶。
Cellulases and related β-1,4-glucanases are essential components of lignocellulose-degrading enzyme mixtures. The detection of β-1,4-glucanase activity typically relies on monitoring the breakdown of purified lignocellulose-derived substrates or synthetic chromogenic substrates, limiting the activities which can be detected and complicating the tracing of activity back to specific components within complex enzyme mixtures. As a tool for the rapid detection and identification of β-1,4-glucanases, a series of glycosylated cyclophellitol inhibitors mimicking β-1,4-glucan oligosaccharides have been synthesised. These compounds are highly efficient inhibitors of HiCel7B, a well-known GH7 endo-β-1,4-glucanase. An elaborated activity-based probe facilitated the direct detection and identification of β-1,4-glucanases within a complex fungal secretome without any detectable cross-reactivity with β-d-glucosidases. These probes and inhibitors add valuable new capacity to the growing toolbox of cyclophellitol-derived probes for the activity-based profiling of biomass-degrading enzymes. New cyclophellitol-derived activity-based probes enable the sensitive detection and identification of cellulases.