Mutation of the eunicellane synthase Bnd4 alters its product profile and expands its prenylation ability.

Mutation of the eunicellane synthase Bnd4 alters its product profile and expands its prenylation ability.
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DOI:
10.1039/d2ob01931k
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发表时间:
2022-11-23
影响因子:
3.2
通讯作者:
--
中科院分区:
化学3区
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--
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Bnd4 催化细菌二萜类双二萜酸生物合成的第一个关键步骤,并且是第一个从自然界中鉴定的单胞烷合酶。我们通过一系列突变研究研究了 Bnd4 活性位点芳香残基的催化作用。这些实验表明,F162 和 Y197 处需要大的疏水性或芳香族侧链来形成单胞菌烷,并且 W316 处的选定突变将 Bnd4 转化为西松烷​​合酶。此外,Bnd4Y197A 变体将 Bnd4 的天然异戊二烯化能力从接受 C5 和 C10 异戊烯基供体扩展到 C15。这项研究支持了 Bnd4 形成 eunicellane 的机制,并鼓励进一步将萜烯合酶改造为实用且高效的异戊二烯基转移酶。
Bnd4 catalyzes the first committed step in the biosynthesis of the bacterial diterpenoid benditerpenoic acid and was the first eunicellane synthase identified from nature. We investigated the catalytic roles of the aromatic residues in the active site of Bnd4 through a series of mutation studies. These experiments revealed that large hydrophobic or aromatic side chains are required at F162 and Y197 for eunicellane formation and that selected mutations at W316 converted Bnd4 into a cembrane synthase. In addition, the Bnd4Y197A variant expanded the native prenylation ability of Bnd4 from accepting C5 and C10 prenyl donors to C15. This study supports the mechanism of eunicellane formation by Bnd4 and encourages further engineering of terpene synthases into practical and efficient prenyltransferases.
DOI: 10.1021/acs.biochem.0c00876
发表时间: 2020-12-22
期刊: Biochemistry
影响因子: 2.9
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