Oligosaccharide hydrolysis by chitosanase enzymes monitored by real-time electrospray ionization-mass spectrometry
Oligosaccharide hydrolysis by chitosanase enzymes monitored by real-time electrospray ionization-mass spectrometry
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DOI：
10.1016/j.jbiotec.2008.02.004
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发表时间：
2008-04-30
影响因子：
4.1
通讯作者：
Letzel, Thomas
中科院分区：
文献类型：
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作者：
Dennhart, Nicole;Fukamizo, Tamo;Letzel, Thomas
首次采用实时电喷雾质谱（ESI-MS）技术研究了壳聚糖酶催化壳寡糖水解反应。利用东方拟无枝酸菌（Amycolatopsis orientalis）的外切壳聚糖酶（exo-beta-glucosaminidase，exo-beta-glucosaminidase）水解壳寡糖（GlcN，n = 2-6），通过将反应溶剂直接注入质谱仪，可同时监测底物、中间体和产物的反应时程。因此，实时MS的分析方法是一种非常节省时间的方法。此外，质谱检测的高灵敏度允许用非常少量的底物以及因此也用少量的应用酶来确定反应时间过程。实时质谱检测也适用于调查链霉菌N174内切壳聚糖酶野生型和它的两个突变体的反应行为。该技术建立了各种酶系统的体外酶活性的快速和有效的测定。(C)2008 Elsevier B. V.保留所有权利。
The chitosanase-catalyzed hydrolysis of chitosan oligosaccharides was investigated for the first time by real-time electrospray ionization-mass spectrometry (ESI-MS). As chitosan oligosaccharides (GlcN, n = 2-6) were hydrolyzed by exochitosanase (exo-beta-glucosaminidase) from Amycolatopsis orientalis, the reaction time-courses of substrate, intermediate and products could be monitored simultaneously by direct infusion of the reaction solvent into the mass spectrometer. Consequently, the analytical approach of real-time MS is an enormous time-saving method. Furthermore, the high sensitivity of the mass spectrometric detection allows the determination of the reaction time-courses with very low quantities of substrate and therefore also a low amount of applied enzyme. Real-time mass spectrometric detection was also applicable in investigating the reaction behaviour of Streptomyces sp. N174 endochitosanase wild type and of two of its mutants. This technique establishes the fast and efficient determination of in vitro enzymatic activities of various enzyme systems. (C) 2008 Elsevier B.V. All rights reserved.