非核糖体肽合成酶中切除的大环化硫酯酶结构域的体外生化特性分析
In Vitro Biochemical Characterization of Excised Macrocyclizing Thioesterase Domains from Non-ribosomal Peptide Synthetases.
作者信息
Brazeau-Henrie Jordan T, Paquette André R, Boddy Christopher N
机构信息
Department of Chemistry and Biomolecular Sciences, University of Ottawa, Ottawa, ON, Canada.
出版信息
Methods Mol Biol. 2023;2670:101-125. doi: 10.1007/978-1-0716-3214-7_5.
Characterization of thioesterases (TEs) is an important step in understanding natural product biosynthesis. Studying non-ribosomal peptide synthetase (NRPS) TEs presents a unique set of challenges with specific cloning and expression issues as well as the challenging synthesis of the thioester peptides substrate required for characterization of the TE. In this method, we describe the cloning and expression of NRPS TEs, the synthesis of thioester peptides, and the in vitro biochemical characterization of the enzyme.
硫酯酶（TEs）的表征是理解天然产物生物合成的重要一步。研究非核糖体肽合成酶（NRPS）的TEs面临着一系列独特的挑战，包括特定的克隆和表达问题，以及表征TE所需的硫酯肽底物的合成难题。在本方法中，我们描述了NRPS TEs的克隆和表达、硫酯肽的合成以及该酶的体外生化表征。
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