4WG2

P411BM3-CIS T438S I263F regioselective C-H amination catalyst

4WG2 の概要

• エントリーDOI: 10.2210/pdb4wg2/pdb
• 関連するPDBエントリー: 4H23
• 分子名称: Bifunctional P-450/NADPH-P450 reductase, SULFATE ION, PROTOPORPHYRIN IX CONTAINING FE, ... (4 entities in total)
• 機能のキーワード: p411bm3-cis, engineering, catalysis, oxidoreductase
• 由来する生物種: Bacillus megaterium
• 細胞内の位置: Cytoplasm : P14779
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 162970.83

構造登録者

Hyster, T.K.,Farwell, C.C.,Buller, A.R.,McIntosh, J.A.,Arnold, F.H. (登録日: 2014-09-17, 公開日: 2014-11-05, 最終更新日: 2023-12-27)

主引用文献

Hyster, T.K.,Farwell, C.C.,Buller, A.R.,McIntosh, J.A.,Arnold, F.H.
Enzyme-controlled nitrogen-atom transfer enables regiodivergent C-h amination.
J.Am.Chem.Soc., 136:15505-15508, 2014

PubMed Abstract: We recently demonstrated that variants of cytochrome P450BM3 (CYP102A1) catalyze the insertion of nitrogen species into benzylic C-H bonds to form new C-N bonds. An outstanding challenge in the field of C-H amination is catalyst-controlled regioselectivity. Here, we report two engineered variants of P450BM3 that provide divergent regioselectivity for C-H amination-one favoring amination of benzylic C-H bonds and the other favoring homo-benzylic C-H bonds. The two variants provide nearly identical kinetic isotope effect values (2.8-3.0), suggesting that C-H abstraction is rate-limiting. The 2.66-Å crystal structure of the most active enzyme suggests that the engineered active site can preorganize the substrate for reactivity. We hypothesize that the enzyme controls regioselectivity through localization of a single C-H bond close to the iron nitrenoid.

PubMed: 25325618
DOI: 10.1021/ja509308v

実験手法

X-RAY DIFFRACTION (2.66 Å)