8QDH

Engineered LmrR carrying a cyclic boronate ester formed between Tris and p-boronophenylalanine at position 89

8QDH の概要

• エントリーDOI: 10.2210/pdb8qdh/pdb
• 分子名称: Transcriptional regulator, PadR-like family, GLYCEROL (3 entities in total)
• 機能のキーワード: artificial enzyme, boron catalysis, unnatural amino acid, 4-boronophenylalanine, transcription
• 由来する生物種: Lactococcus cremoris subsp. cremoris MG1363
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 30822.19

構造登録者

Thunnissen, A.M.W.H.,Rozeboom, H.J.,Longwitz, L.,Leveson-Gower, R.B.,Roelfes, G. (登録日: 2023-08-29, 公開日: 2024-05-01, 最終更新日: 2024-06-05)

主引用文献

Longwitz, L.,Leveson-Gower, R.B.,Rozeboom, H.J.,Thunnissen, A.W.H.,Roelfes, G.
Boron catalysis in a designer enzyme.
Nature, 629:824-829, 2024

PubMed Abstract: Enzymes play an increasingly important role in improving the benignity and efficiency of chemical production, yet the diversity of their applications lags heavily behind chemical catalysts as a result of the relatively narrow range of reaction mechanisms of enzymes. The creation of enzymes containing non-biological functionalities facilitates reaction mechanisms outside nature's canon and paves the way towards fully programmable biocatalysis. Here we present a completely genetically encoded boronic-acid-containing designer enzyme with organocatalytic reactivity not achievable with natural or engineered biocatalysts. This boron enzyme catalyses the kinetic resolution of hydroxyketones by oxime formation, in which crucial interactions with the protein scaffold assist in the catalysis. A directed evolution campaign led to a variant with natural-enzyme-like enantioselectivities for several different substrates. The unique activation mode of the boron enzyme was confirmed using X-ray crystallography, high-resolution mass spectrometry (HRMS) and B NMR spectroscopy. Our study demonstrates that genetic-code expansion can be used to create evolvable enantioselective enzymes that rely on xenobiotic catalytic moieties such as boronic acids and access reaction mechanisms not reachable through catalytic promiscuity of natural or engineered enzymes.

PubMed: 38720081
DOI: 10.1038/s41586-024-07391-3

実験手法

X-RAY DIFFRACTION (1.72 Å)