Abstract：
Determining enzyme activities involved in photorespiration, either in a crude plant tissue extract or in a preparation of a recombinant enzyme, is time-consuming, especially when large number of samples need to be processed. This chapter presents a phosphoglycolate phosphatase (PGLP) activity assay that is adapted for use in a 96-well microplate format. The microplate format for the assay requires fewer enzymes and reagents and allows rapid and less expensive measurement of PGLP enzyme activity. The small volume of reaction mix in a 96-well microplate format enables the determination of PGLP enzyme activity for screening many plant samples, multiple enzyme activities using the same protein extract, and/or identifying kinetic parameters for a recombinant enzyme. To assist in preparing assay reagents, we also present an R Shiny buffer preparation app for PGLP and other photorespiratory enzyme activities and a Km and Vmax calculation app.
摘要：
确定光呼吸中涉及的酶活性，在粗植物组织提取物或重组酶的制备中，是耗时的，特别是当需要处理大量样品时。本章介绍了磷酸葡萄糖酸磷酸酶（PGLP）活性测定，适用于96孔微孔板格式。用于测定的微孔板形式需要较少的酶和试剂,并且允许快速且较便宜地测量PGLP酶活性。96孔微孔板形式的小体积反应混合物能够测定用于筛选许多植物样品的PGLP酶活性,使用相同的蛋白质提取物的多种酶活性，和/或鉴定重组酶的动力学参数。为了协助制备测定试剂，我们还提出了一个RShiny缓冲液制备应用程序，用于PGLP和其他光呼吸酶活性以及Km和Vmax计算应用程序。
