Department of Chemistry, Ludwig-Maximilians-Universität München, München, Germany.
Department of Chemistry, Ludwig-Maximilians-Universität München, München, Germany.
Methods Enzymol. 2021;650:57-79. doi: 10.1016/bs.mie.2021.01.045. Epub 2021 Mar 4.
The field of methanol dehydrogenases (MDHs) has experienced revival in the recent decade due to the observation of lanthanide-dependent MDH, in addition to widely known calcium-MDH. With the advent of lanthanide-dependent alcohol dehydrogenases, the need for reliable assays to evaluate and compare activities between different MDHs is obvious: from extremophilic to neutrophilic organisms, or with different lanthanide ions in the active site. Here we outline four assays that have been reported for Ln-MDH, discussing the advantages and disadvantages of the assays and their components. It should be noted, in 1990Day and Anthony produced a comprehensive summary in Methods in Enzymology on the available methods for Ca-MDH assays at the time (Day & Anthony, 1990). This chapter is an updated appraisal of the most important developments in the last 30years.
甲醇脱氢酶(MDH)领域在最近十年中经历了复兴,这是由于观察到了除了广为人知的钙-MDH 之外,还存在镧系依赖性 MDH。随着镧系依赖性醇脱氢酶的出现,显然需要可靠的测定法来评估和比较不同 MDH 之间的活性:从极端嗜热生物体到嗜中性生物体,或者在活性部位具有不同的镧系离子。在这里,我们概述了已报道的用于 Ln-MDH 的四种测定法,讨论了测定法及其组成部分的优缺点。需要注意的是,1990 年 Day 和 Anthony 在《酶学方法》(Methods in Enzymology)中全面总结了当时用于 Ca-MDH 测定的可用方法(Day & Anthony,1990)。本章是对过去 30 年中最重要进展的最新评价。