8SU2

F96H epi-Isozizaene Synthase: complex with 3 Mg2+ and risedronate

8SU2 の概要

• エントリーDOI: 10.2210/pdb8su2/pdb
• 分子名称: Epi-isozizaene synthase, SULFATE ION, GLYCEROL, ... (6 entities in total)
• 機能のキーワード: terpene, terpenoid, isoprenoid, terpene cyclase, terpene synthase, lyase
• 由来する生物種: Streptomyces coelicolor A3(2)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 44251.17

構造登録者

Eaton, S.A.,Christianson, D.W. (登録日: 2023-05-11, 公開日: 2023-07-26, 最終更新日: 2023-08-16)

主引用文献

Eaton, S.A.,Christianson, D.W.
Reprogramming the Cyclization Cascade of epi -Isozizaene Synthase to Generate Alternative Terpene Products.
Biochemistry, 62:2301-2313, 2023

PubMed Abstract: The class I sesquiterpene cyclase -isozizaene synthase from (EIZS) catalyzes the transformation of linear farnesyl diphosphate (FPP) into the tricyclic hydrocarbon -isozizaene in the biosynthesis of albaflavenone antibiotics. The active site cavity of EIZS is largely framed by four aromatic residues - F95, F96, F198, and W203 - that form a product-shaped contour, serving as a template to chaperone conformations of the flexible substrate and multiple carbocation intermediates leading to -isozizaene. Remolding the active site contour by mutagenesis can redirect the cyclization cascade away from -isozizaene biosynthesis to generate alternative sesquiterpene products. Here, we present the biochemical and structural characterization of four EIZS mutants in which aromatic residues have been substituted with polar residues (F95S, F96H, F198S, and F198T) to generate alternative cyclization products. Most notably, F95S EIZS generates a mixture of monocyclic sesquiterpene precursors of bisabolane, a D2 diesel fuel substitute. X-ray crystal structures of the characterized mutants reveal subtle changes in the active site contour showing how each aromatic residue influences the chemistry of a different carbocation intermediate in the cyclization cascade. We advance that EIZS may serve as a robust platform for the development of designer cyclases for the generation of high-value sesquiterpene products ranging from pharmaceuticals to biofuels in synthetic biology approaches.

PubMed: 37449555
DOI: 10.1021/acs.biochem.3c00247

実験手法

X-RAY DIFFRACTION (1.33 Å)