Horn Svein J, Sørlie Morten, Vårum Kjell M, Väljamäe Priit, Eijsink Vincent G H
Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Aas, Norway.
Methods Enzymol. 2012;510:69-95. doi: 10.1016/B978-0-12-415931-0.00005-7.
Natural cellulolytic enzyme systems as well as leading commercial cellulase cocktails are dominated by enzymes that degrade cellulose chains in a processive manner. Despite the abundance of processivity among natural cellulases, the molecular basis as well as the biotechnological implications of this mechanism are only partly understood. One of the major limitations lies in the fact that it is not straightforward to measure and quantify processivity in what essentially are biphasic experimental systems. Here, we describe and discuss both well-established methods and newer methods for measuring cellulase processivity. In addition, we discuss recent insights from studies on chitinases that may help direct further studies on processivity in cellulases.
天然纤维素分解酶系统以及主要的商业纤维素酶混合物都以能够连续降解纤维素链的酶为主。尽管天然纤维素酶中存在大量的连续性,但这种机制的分子基础以及生物技术意义仍仅得到部分理解。主要限制之一在于,在本质上是双相实验系统中测量和量化连续性并非易事。在此,我们描述并讨论了用于测量纤维素酶连续性的既定方法和新方法。此外,我们还讨论了来自几丁质酶研究的最新见解,这些见解可能有助于指导对纤维素酶连续性的进一步研究。