Heterologous Expression, Purification, and Characterization of Type II Polyketide Synthase Acyl Carrier Proteins.
Heterologous Expression, Purification, and Characterization of Type II Polyketide Synthase Acyl Carrier Proteins.
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DOI：
10.1007/978-1-0716-2273-5_13
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发表时间：
2022
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包含II型聚酮化合物脱氢酶（PKS）的酶是强大的生物催化剂，一旦被充分理解和战略性应用，就可以实现对一系列药学相关分子的成本有效和可持续的获取。实现这一目标的进展取决于获得足够的材料，在体外表征和结构分析的组成部分，这些酶。PKSs的一个中心组分是酰基载体蛋白（ACP），其在II型聚酮化合物的生物合成过程中充当枢纽。在此，我们分享了通过在大肠杆菌中异源表达获得II型PKS ACP的方法。杆菌我们还分享了如何安装反应性和位点特异性光谱探针可以利用来研究II型PKS ACP的构象动力学和相互作用。
The enzymes that comprise type II polyketide synthases (PKSs) are powerful biocatalysts that, once well-understood and strategically applied, could enable cost-effective and sustainable access to a range of pharmaceutically relevant molecules. Progress towards this goal hinges on gaining ample access to materials for in vitro characterizations and structural analyses of the components of these synthases. A central component of PKSs is the acyl carrier protein (ACP), which serves as a hub during the biosynthesis of type II polyketides. Herein, we share methods for accessing type II PKS ACPs via heterologous expression in E. coli. We also share how the installation of reactive and site-specific spectroscopic probes can be leveraged to study the conformational dynamics and interactions of type II PKS ACPs.