6SDM

NADH-dependent variant of TBADH

6SDM の概要

• エントリーDOI: 10.2210/pdb6sdm/pdb
• 分子名称: NADP-dependent isopropanol dehydrogenase, ZINC ION (2 entities in total)
• 機能のキーワード: oxidoreductase, cofactor
• 由来する生物種: Thermoanaerobacter brockii
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 151910.53

構造登録者

Selles Vidal, L.,Murray, J.W.,Heap, J.T. (登録日: 2019-07-28, 公開日: 2020-08-26, 最終更新日: 2024-01-24)

主引用文献

Selles Vidal, L.,Murray, J.W.,Heap, J.T.
Versatile selective evolutionary pressure using synthetic defect in universal metabolism.
Nat Commun, 12:6859-6859, 2021

PubMed Abstract: The non-natural needs of industrial applications often require new or improved enzymes. The structures and properties of enzymes are difficult to predict or design de novo. Instead, semi-rational approaches mimicking evolution entail diversification of parent enzymes followed by evaluation of isolated variants. Artificial selection pressures coupling desired enzyme properties to cell growth could overcome this key bottleneck, but are usually narrow in scope. Here we show diverse enzymes using the ubiquitous cofactors nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP) can substitute for defective NAD regeneration, representing a very broadly-applicable artificial selection. Inactivation of Escherichia coli genes required for anaerobic NAD regeneration causes a conditional growth defect. Cells are rescued by foreign enzymes connected to the metabolic network only via NAD or NADP, but only when their substrates are supplied. Using this principle, alcohol dehydrogenase, imine reductase and nitroreductase variants with desired selectivity modifications, and a high-performing isopropanol metabolic pathway, are isolated from libraries of millions of variants in single-round experiments with typical limited information to guide design.

PubMed: 34824282
DOI: 10.1038/s41467-021-27266-9

実験手法

X-RAY DIFFRACTION (2.85 Å)