Srividya N, Lange I, Lange B M
Institute of Biological Chemistry, M. J. Murdock Metabolomics Laboratory, Washington State University, Pullman, WA, United States.
Institute of Biological Chemistry, M. J. Murdock Metabolomics Laboratory, Washington State University, Pullman, WA, United States.
Methods Enzymol. 2016;576:147-65. doi: 10.1016/bs.mie.2016.03.013. Epub 2016 Mar 28.
Monoterpene synthases are highly versatile enzymes that catalyze the first committed step in the pathways toward terpenoids, the structurally most diverse class of plant natural products. Recent advancements in our understanding of the reaction mechanism have enabled engineering approaches to develop mutant monoterpene synthases that produce specific monoterpenes. In this chapter, we are describing protocols to introduce targeted mutations, express mutant enzyme catalysts in heterologous hosts, and assess their catalytic properties. Mutant monoterpene synthases have the potential to contribute significantly to synthetic biology efforts aimed at producing larger amounts of commercially attractive monoterpenes.
单萜合酶是高度通用的酶，可催化通往萜类化合物途径中的首个关键步骤，萜类化合物是植物天然产物中结构最多样化的一类。我们对反应机制理解的最新进展使得能够采用工程方法来开发产生特定单萜的突变单萜合酶。在本章中，我们将描述引入靶向突变、在异源宿主中表达突变酶催化剂以及评估其催化特性的方案。突变单萜合酶有潜力为旨在大量生产具有商业吸引力的单萜的合成生物学努力做出重大贡献。
