7F3P

Crystal structure of a nadp-dependent alcohol dehydrogenase mutant in apo form

7F3P の概要

• エントリーDOI: 10.2210/pdb7f3p/pdb
• 分子名称: NADP-dependent isopropanol dehydrogenase, ZINC ION, NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, ... (4 entities in total)
• 機能のキーワード: alpha/beta dehydrogenase, oxidoreductase
• 由来する生物種: Thermoanaerobacter brockii (Thermoanaerobium brockii)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 155059.90

構造登録者

Han, X.,Bi, Y.,Wei, H.L.,Gao, J.,Li, Q.,Qu, G.,Sun, Z.T.,Liu, W.D. (登録日: 2021-06-16, 公開日: 2021-10-27, 最終更新日: 2023-11-29)

主引用文献

Qu, G.,Bi, Y.,Liu, B.,Li, J.,Han, X.,Liu, W.,Jiang, Y.,Qin, Z.,Sun, Z.
Unlocking the Stereoselectivity and Substrate Acceptance of Enzymes: Proline-Induced Loop Engineering Test.
Angew.Chem.Int.Ed.Engl., 61:e202110793-e202110793, 2022

PubMed Abstract: Protein stability and evolvability influence each other. Although protein dynamics play essential roles in various catalytically important properties, their high flexibility and diversity makes it difficult to incorporate such properties into rational engineering. Therefore, how to unlock the potential evolvability in a user-friendly rational design process remains a challenge. In this endeavor, we describe a method for engineering an enantioselective alcohol dehydrogenase. It enables synthetically important substrate acceptance for 4-chlorophenyl pyridine-2-yl ketone, and perfect stereocontrol of both (S)- and (R)-configured products. Thermodynamic analysis unveiled the subtle interaction between enzyme stability and evolvability, while computational studies provided insights into the origin of selectivity and substrate recognition. Preparative-scale synthesis of the (S)-product (73 % yield; >99 % ee) was performed on a gram-scale. This proof-of-principle study demonstrates that interfaced proline residues can be rationally engineered to unlock evolvability and thus provide access to new biocatalysts with highly improved catalytic performance.

PubMed: 34658118
DOI: 10.1002/anie.202110793

実験手法

X-RAY DIFFRACTION (2.6 Å)