An ultrasensitive, continuous assay for xylanase using the fluorogenic substrate 6,8-difluoro-4-methylumbelliferyl β-D-xylobloside
An ultrasensitive, continuous assay for xylanase using the fluorogenic substrate 6,8-difluoro-4-methylumbelliferyl β-D-xylobloside
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DOI：
10.1016/j.ab.2006.11.034
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发表时间：
2007-03-01
影响因子：
2.9
通讯作者：
Johnson, Lain
中科院分区：
文献类型：
--
作者：
Ge, Yue;Antoulinakis, Evan G.;Johnson, Lain
我们描述了一种新的荧光底物--6，8-二氟-4-甲基伞形花环基β-D-木糖苷(DiFMUX(2))用于木聚糖酶活性的荧光分析。底物中6，8-二氟-4-甲基伞形酮(DiFMU)的产生直接与木聚糖酶活性相对应。高效液相色谱分析表明，木聚糖酶将DiFMUX2直接降解为DiFMU和木二糖。该方法具有测量木聚糖酶活性或高通量筛选木聚糖酶抑制剂所需的速度、灵敏度和方便性，适用于各种来源的木聚糖酶的动力学分析。(C)2006 Elsevier Inc.保留所有权利。
We describe a fluorescence-based assay for the analysis of xylanase activity using a novel fluorogenic substrate, 6,8-difluoro-4-methylumbelliferyl beta-D-Xylobioside (DiFMUX(2)). Generation of fluorescent 6,8-difluoro-4-methylumbelliferone (DiFMU) from the substrate corresponded directly to xylanase activity. High-performance liquid chromatography analysis of the digestion products showed that xylanase hydrolyzed DiFMUX2 directly to DiFMU and xylobiose. The assay provides the speed, sensitivity, and convenience required for measuring xylanase activity or for screening xylanase inhibitors in a high-throughput format and is suitable for the kinetic assay of xylanases from a variety of sources. (c) 2006 Elsevier Inc. All rights reserved.