Protocol for detecting threonine deaminase activity in fission yeast cell lysates

Threonine deaminase catalyzes the first step of isoleucine biosynthesis from threonine. In this protocol, we describe the process of measuring the enzymatic activity of threonine deaminase in the fission yeast cell lysate, which is catalyzed by Tda1. First, we describe the process of preparing cell lysates from fission yeast cell cultures. Subsequently, we explain how to measure the threonine deaminase activity using threonine or serine as a substrate.For complete details on the use and execution of this protocol, please refer to Sasaki et al. (2022).1

Automated summary

This protocol describes the process of synthesizing isoleucine from threonine by measuring the enzymatic activity of threonine deaminase in fission yeast cell lysate. The enzymatic activity is catalyzed by Tda1. The protocol involves preparing cell lysates from fission yeast cell cultures and measuring threonine deaminase activity using threonine or serine as a substrate.