4OQS

Crystal structure of CYP105AS1

4OQS の概要

• エントリーDOI: 10.2210/pdb4oqs/pdb
• 関連するPDBエントリー: 4OQR
• 分子名称: CYP105AS1, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total)
• 機能のキーワード: cytochrome p450, monooxygenase, oxidoreductase
• 由来する生物種: Amycolatopsis orientalis
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 48624.38

構造登録者

Leys, D. (登録日: 2014-02-10, 公開日: 2015-02-18, 最終更新日: 2023-09-20)

主引用文献

McLean, K.J.,Hans, M.,Meijrink, B.,van Scheppingen, W.B.,Vollebregt, A.,Tee, K.L.,van der Laan, J.M.,Leys, D.,Munro, A.W.,van den Berg, M.A.
Single-step fermentative production of the cholesterol-lowering drug pravastatin via reprogramming of Penicillium chrysogenum.
Proc.Natl.Acad.Sci.USA, 112:2847-2852, 2015

PubMed Abstract: The cholesterol-lowering blockbuster drug pravastatin can be produced by stereoselective hydroxylation of the natural product compactin. We report here the metabolic reprogramming of the antibiotics producer Penicillium chrysogenum toward an industrial pravastatin production process. Following the successful introduction of the compactin pathway into the β-lactam-negative P. chrysogenum DS50662, a new cytochrome P450 (P450 or CYP) from Amycolatopsis orientalis (CYP105AS1) was isolated to catalyze the final compactin hydroxylation step. Structural and biochemical characterization of the WT CYP105AS1 reveals that this CYP is an efficient compactin hydroxylase, but that predominant compactin binding modes lead mainly to the ineffective epimer 6-epi-pravastatin. To avoid costly fractionation of the epimer, the enzyme was evolved to invert stereoselectivity, producing the pharmacologically active pravastatin form. Crystal structures of the optimized mutant P450(Prava) bound to compactin demonstrate how the selected combination of mutations enhance compactin binding and enable positioning of the substrate for stereo-specific oxidation. Expression of P450(Prava) fused to a redox partner in compactin-producing P. chrysogenum yielded more than 6 g/L pravastatin at a pilot production scale, providing an effective new route to industrial scale production of an important drug.

PubMed: 25691737
DOI: 10.1073/pnas.1419028112

実験手法

X-RAY DIFFRACTION (2.04 Å)