8AQ0

Crystal structure of L-N-Carbamoylase from Sinorhizobium meliloti mutant L217G/F329C

8AQ0 の概要

• エントリーDOI: 10.2210/pdb8aq0/pdb
• 関連するPDBエントリー: 8APZ
• 分子名称: N-carbamoyl-L-amino-acid hydrolase, ZINC ION, FE (III) ION, ... (6 entities in total)
• 機能のキーワード: peptidase family m20/m25/m40, hydrolase
• 由来する生物種: Sinorhizobium meliloti
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 94847.11

構造登録者

Rozeboom, H.J.,Mayer, C. (登録日: 2022-08-11, 公開日: 2022-11-16, 最終更新日: 2024-05-01)

主引用文献

Rubini, R.,Jansen, S.C.,Beekhuis, H.,Rozeboom, H.J.,Mayer, C.
Selecting Better Biocatalysts by Complementing Recoded Bacteria.
Angew.Chem.Int.Ed.Engl., 62:e202213942-e202213942, 2023

PubMed Abstract: In vivo selections are powerful tools for the directed evolution of enzymes. However, the need to link enzymatic activity to cellular survival makes selections for enzymes that do not fulfill a metabolic function challenging. Here, we present an in vivo selection strategy that leverages recoded organisms addicted to non-canonical amino acids (ncAAs) to evolve biocatalysts that can provide these building blocks from synthetic precursors. We exemplify our platform by engineering carbamoylases that display catalytic efficiencies more than five orders of magnitude higher than those observed for the wild-type enzyme for ncAA-precursors. As growth rates of bacteria under selective conditions correlate with enzymatic activities, we were able to elicit improved variants from populations by performing serial passaging. By requiring minimal human intervention and no specialized equipment, we surmise that our strategy will become a versatile tool for the in vivo directed evolution of diverse biocatalysts.

PubMed: 36342942
DOI: 10.1002/anie.202213942

実験手法

X-RAY DIFFRACTION (2.3 Å)