Controlling Chemo- and Regioselectivity of a Plant P450 in Yeast Cell toward Rare Licorice Triterpenoid Biosynthesis,ACS Catalysis

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Controlling Chemo- and Regioselectivity of a Plant P450 in Yeast Cell toward Rare Licorice Triterpenoid Biosynthesis
ACS Catalysis ( IF 11.3 ) Pub Date : 2020-03-20 , DOI: 10.1021/acscatal.0c00128
Wentao Sun ₁ , Haijie Xue ₁ , Hu Liu ₁ , Bo Lv ₁ , Yang Yu ₁ , Ying Wang ₁ , Meilan Huang ₂ , Chun Li _{1,

3}

Affiliation

The hallmark reaction of P450 monooxygenase involves activation of C–H bond and the production of a hydroxyl. P450s tailoring natural product could further oxidize the hydroxyl to carboxylic acid. However, heterogeneously expressed plant P450s display poor chemo- and regioselectivity in microbes, restricting the efficient biosynthesis of related natural products. CYP72A63 is a P450 enzyme previously used for the biosynthesis of glycyrrhetinic acid, and its poor selectivity resulted in oxidation of 11-oxo-β-amyrin to a mixture of rare licorice triterpenoids (glycyrrhetol, glycyrrhetaldehyde, glycyrrhetinic acid, and 29-OH-11-oxo-β-amyrin). In this study, we have identified key residues, which influence the enzyme–substrate hydrophobic interaction, in controlling the chemo- and regioselectivity of the enzyme and engineered the enzyme toward selectivity oxidation to hydroxyl and carboxylic acid. Moreover, tuning the redox partner of the P450 leads to selective production of glycyrrhetaldehyde, a good starting point for further modification. In this study, controlling the catalytic property of plant P450s prove to be of great use in the synthesis of desired licorice triterpenoids, which can be used in biosynthesis of other terpenoid natural products.

中文翻译：

控制酵母细胞中植物P450对稀有甘草三萜类生物合成的化学和区域选择性

P450单加氧酶的标志性反应涉及C–H键的活化和羟基的产生。定制天然产物的P450可以进一步将羟基氧化为羧酸。但是，异源表达的植物P450在微生物中显示出较差的化学和区域选择性，从而限制了相关天然产物的有效生物合成。CYP72A63是一种P450酶，以前曾用于生物合成甘草次酸，其选择性差，导致11-氧代-β-淀粉糊精氧化成稀有甘草三萜类化合物（甘草醇，甘草醛，甘草次酸和29-OH-11 -氧代-β-amyrin）。在这项研究中，我们确定了影响酶-底物疏水相互作用的关键残基，在控制酶的化学和区域选择性上，并设计了酶使其选择性氧化成羟基和羧酸。此外，调整P450的氧化还原配偶体会导致选择性生产甘草次醛，这是进一步修饰的良好起点。在这项研究中，控制植物P450的催化性能被证明在合成所需的甘草三萜类化合物中具有很大的用途，可用于其他萜类天然产物的生物合成。

更新日期：2020-03-21

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