Discovery and Biocatalytic Application of a PLP-Dependent Amino Acid γ-Substitution Enzyme that Catalyzes C-C Bond Formation,Journal of the American Chemical Society

当前位置： X-MOL 学术 › J. Am. Chem. Soc. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Discovery and Biocatalytic Application of a PLP-Dependent Amino Acid γ-Substitution Enzyme that Catalyzes C-C Bond Formation
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-05-20 , DOI: 10.1021/jacs.0c03535
Mengbin Chen ₁ , Chun-Ting Liu _{1,

2} , Yi Tang _{1,

3}

Affiliation

Pyridoxal phosphate (PLP)-dependent enzymes can catalyze transformations of L-amino acids at α, β and γ positions. These enzymes are frequently involved in the biosynthesis of nonproteinogenic amino acids as building blocks of natural products, and are attractive biocatalysts. Here, we report the discovery of a two-step enzymatic synthesis of (2S, 6S)-6-methyl pipecolate 1, from the biosynthetic pathway of citrinadin. The key enzyme CndF is PLP-dependent and catalyzes synthesis of (S)-2-amino-6-oxoheptanoate 3 that is in equilibrium with the cyclic Schiff base. The second enzyme CndE is a stereoselective imine reductase that gives 1. Biochemical characterization of CndF showed this enzyme performs γ-elimination of O-acetyl-L-homoserine to generate the vinylglycine ketimine, which is subjected to nucleophilic attack by acetoacetate to form the new Cγ-Cδ bond in 3 and complete the γ-substitution reaction. CndF displays promiscuity towards different β-keto carboxylate and esters. Using an Aspergillus strain expressing CndF and CndE, feeding various alkyl-β-keto esters led to the biosynthesis of 6-substituted L-pipecolates. The discovery of CndF expands the repertoire of reactions that can be catalyzed by PLP-dependent enzymes.

中文翻译：

一种催化 CC 键形成的 PLP 依赖性氨基酸 γ-取代酶的发现和生物催化应用

磷酸吡哆醛 (PLP) 依赖性酶可以催化 L-氨基酸在 α、β 和 γ 位点的转化。这些酶经常参与作为天然产物构建块的非蛋白氨基酸的生物合成，并且是有吸引力的生物催化剂。在这里，我们报告了 (2S, 6S)-6-methyl pipecolate 1 的两步酶法合成的发现，来自 citrinadin 的生物合成途径。关键酶 CndF 依赖于 PLP，可催化 (S)-2-amino-6-oxoheptanoate 3 的合成，该化合物与环状 Schiff 碱处于平衡状态。第二种酶 CndE 是一种立体选择性亚胺还原酶，可产生 1。CndF 的生化表征表明该酶执行 O-乙酰基-L-高丝氨酸的 γ-消除以生成乙烯基甘氨酸酮亚胺，其受到乙酰乙酸酯的亲核攻击以在3中形成新的Cγ-Cδ键并完成γ-取代反应。CndF 对不同的 β-酮羧酸酯和酯表现出混杂性。使用表达 CndF 和 CndE 的曲霉菌株，喂食各种烷基-β-酮酯导致 6-取代的 L-哌可酯的生物合成。CndF 的发现扩展了可由 PLP 依赖性酶催化的反应库。

更新日期：2020-05-20

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文本刊介绍/投稿指南