3P67

T26S mutant of pentaerythritol tetranitrate reductase containing a bound acetate molecule

3P67 の概要

• エントリーDOI: 10.2210/pdb3p67/pdb
• 関連するPDBエントリー: 1H50 3P62
• 分子名称: Pentaerythritol tetranitrate reductase, FLAVIN MONONUCLEOTIDE, ACETATE ION, ... (4 entities in total)
• 機能のキーワード: old yellow enzyme family, alpha, beta barrel, oxidoreductase
• 由来する生物種: Enterobacter cloacae
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 41141.72

構造登録者

Toogood, H.S.,Scrutton, N.S. (登録日: 2010-10-11, 公開日: 2010-11-24, 最終更新日: 2023-11-01)

主引用文献

Hulley, M.E.,Toogood, H.S.,Fryszkowska, A.,Mansell, D.,Stephens, G.M.,Gardiner, J.M.,Scrutton, N.S.
Focused Directed Evolution of Pentaerythritol Tetranitrate Reductase by Using Automated Anaerobic Kinetic Screening of Site-Saturated Libraries
Chembiochem, 11:2433-2447, 2010

PubMed Abstract: This work describes the development of an automated robotic platform for the rapid screening of enzyme variants generated from directed evolution studies of pentraerythritol tetranitrate (PETN) reductase, a target for industrial biocatalysis. By using a 96-well format, near pure enzyme was recovered and was suitable for high throughput kinetic assays; this enabled rapid screening for improved and new activities from libraries of enzyme variants. Initial characterisation of several single site-saturation libraries targeted at active site residues of PETN reductase, are described. Two mutants (T26S and W102F) were shown to have switched in substrate enantiopreference against substrates (E)-2-aryl-1-nitropropene and α-methyl-trans-cinnamaldehyde, respectively, with an increase in ee (62 % (R) for W102F). In addition, the detection of mutants with weak activity against α,β-unsaturated carboxylic acid substrates showed progress in the expansion of the substrate range of PETN reductase. These methods can readily be adapted for rapid evolution of enzyme variants with other oxidoreductase enzymes.

PubMed: 21064170
DOI: 10.1002/cbic.201000527

実験手法

X-RAY DIFFRACTION (1.5 Å)