FAD-dependent enzyme-catalysed intermolecular [4+2] cycloaddition in natural product biosynthesis

被引：124

作者：

Gao, Lei ^{[1
,2
,3
,4
,5
]}

Su, Cong ^{[1
,2
,3
]}

Du, Xiaoxia ^{[4
,5
]}

Wang, Ruishan ^{[6
]}

Chen, Shuming ^{[7
]}

Zhou, Yu ^{[8
]}

Liu, Chengwei ^{[9
]}

Liu, Xiaojing ^{[4
,5
]}

Tian, Runze ^{[4
,5
]}

Zhang, Liyun ^{[4
,5
]}

Xie, Kebo ^{[1
,2
,3
]}

Chen, She ^{[8
]}

Guo, Qianqian ^{[4
,5
]}

Guo, Lanping ^{[6
]}

Hano, Yoshio ^{[10
]}

Shimazaki, Manabu ^{[10
]}

Minami, Atsushi ^{[9
]}

Oikawa, Hideaki ^{[9
]}

Huang, Niu ^{[8
]}

Houk, K. N. ^{[7
]}

Huang, Luqi ^{[6
]}

Dai, Jungui ^{[1
,2
,3
]}

Lei, Xiaoguang ^{[4
,5
]}

机构：

[1] State Key Lab Bioact Subst & Funct Nat Med, Beijing, Peoples R China

[2] Chinese Acad Med Sci, CAMS Key Lab Enzyme & Biocatalysis Nat Drugs, Inst Mat Med, Beijing, Peoples R China

[3] Peking Union Med Coll, Beijing, Peoples R China

[4] Peking Univ, Beijing Natl Lab Mol Sci, Key Lab Bioorgan Chem & Mol Engn, Minist Educ,Dept Chem Biol,Coll Chem & Mol Engn,S, Beijing, Peoples R China

[5] Peking Univ, Peking Tsinghua Ctr Life Sci, Beijing, Peoples R China

[6] China Acad Chinese Med Sci, Natl Resources Ctr Chinese Mat Med, State Key Lab Breeding Base Dao Di Herbs, Beijing, Peoples R China

[7] Univ Calif Los Angeles, Dept Chem & Biochem, 405 Hilgard Ave, Los Angeles, CA 90024 USA

[8] Natl Inst Biol Sci NIBS, Beijing, Peoples R China

[9] Hokkaido Univ, Grad Sch Sci, Div Chem, Sapporo, Hokkaido, Japan

[10] Teikyo Heisei Univ, Fac Pharmaceut Sci, Tokyo, Japan

来源：

NATURE CHEMISTRY | 2020年 / 12卷 / 07期

基金：

美国国家科学基金会; 中国国家自然科学基金;

关键词：

DIELS-ALDER REACTION; PERICYCLIC-REACTIONS; KUWANON-J; ADDUCTS; SYNTHASE; IDENTIFICATION; CHALCOMORACIN; CYCLIZATION; BROSIMONES; CHEMISTRY;

D O I：

10.1038/s41557-020-0467-7

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

The Diels-Alder reaction is one of the most powerful and widely used methods in synthetic chemistry for the stereospecific construction of carbon-carbon bonds. Despite the importance of Diels-Alder reactions in the biosynthesis of numerous secondary metabolites, no naturally occurring stand-alone Diels-Alderase has been demonstrated to catalyse intermolecular Diels-Alder transformations. Here we report a flavin adenine dinucleotide-dependent enzyme, Morusalba Diels-Alderase (MaDA), from Morus cell cultures, that catalyses an intermolecular [4+2] cycloaddition to produce the natural isoprenylated flavonoid chalcomoracin with a high efficiency and enantioselectivity. Density functional theory calculations and preliminary measurements of the kinetic isotope effects establish a concerted but asynchronous pericyclic pathway. Structure-guided mutagenesis and docking studies demonstrate the interactions of MaDA with the diene and dienophile to catalyse the [4+2] cycloaddition. MaDA exhibits a substrate promiscuity towards both dienes and dienophiles, which enables the expedient syntheses of structurally diverse natural products. We also report a biosynthetic intermediate probe (BIP)-based target identification strategy used to discover MaDA.

引用

页码：620 / +

页数：26

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