7ROF

Engineered tryptophan synthase b-subunit from Pyrococcus furiosus, PfTrpB2B9-H275E with L-Trp non-covalently bound

7ROF の概要

• エントリーDOI: 10.2210/pdb7rof/pdb
• 関連するPDBエントリー: 7RNP 7RNQ
• 分子名称: Tryptophan synthase beta chain 1, TRYPTOPHAN, SODIUM ION, ... (4 entities in total)
• 機能のキーワード: tryptophan synthase, biosynthetic protein
• 由来する生物種: Pyrococcus furiosus
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 174939.16

構造登録者

Higgins, P.M.,Buller, A.R. (登録日: 2021-07-30, 公開日: 2022-08-03, 最終更新日: 2023-11-15)

主引用文献

McDonald, A.D.,Higgins, P.M.,Buller, A.R.
Substrate multiplexed protein engineering facilitates promiscuous biocatalytic synthesis.
Nat Commun, 13:5242-5242, 2022

PubMed Abstract: Enzymes with high activity are readily produced through protein engineering, but intentionally and efficiently engineering enzymes for an expanded substrate scope is a contemporary challenge. One approach to address this challenge is Substrate Multiplexed Screening (SUMS), where enzyme activity is measured on competing substrates. SUMS has long been used to rigorously quantitate native enzyme specificity, primarily for in vivo settings. SUMS has more recently found sporadic use as a protein engineering approach but has not been widely adopted by the field, despite its potential utility. Here, we develop principles of how to design and interpret SUMS assays to guide protein engineering. This rich information enables improving activity with multiple substrates simultaneously, identifies enzyme variants with altered scope, and indicates potential mutational hot-spots as sites for further engineering. These advances leverage common laboratory equipment and represent a highly accessible and customizable method for enzyme engineering.

PubMed: 36068220
DOI: 10.1038/s41467-022-32789-w

実験手法

X-RAY DIFFRACTION (2.39 Å)