7N2T

O-acetylserine sulfhydrylase from Citrullus vulgaris in the internal aldimine state, with citrate bound

7N2T の概要

• エントリーDOI: 10.2210/pdb7n2t/pdb
• 分子名称: Cysteine synthase, CITRIC ACID, PENTAETHYLENE GLYCOL, ... (5 entities in total)
• 機能のキーワード: o-acetylserine sulfhydrylase, plp, cysteine synthase, biosynthetic protein
• 由来する生物種: Citrullus lanatus subsp. vulgaris (watermelon)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 36204.26

構造登録者

Smith, J.L.,Buller, A.R.,Bingman, C.A. (登録日: 2021-05-29, 公開日: 2022-07-06, 最終更新日: 2024-04-03)

主引用文献

Smith, J.L.,Harrison, I.M.,Bingman, C.A.,Buller, A.R.
Investigation of beta-Substitution Activity of O-Acetylserine Sulfhydrolase from Citrullus vulgaris.
Chembiochem, 23:e202200157-e202200157, 2022

PubMed Abstract: Pyridoxal-5'-phosphate (PLP)-dependent enzymes have garnered interest for their ability to synthesize non-standard amino acids (nsAAs). One such class of enzymes, O-acetylserine sulfhydrylases (OASSs), catalyzes the final step in the biosynthesis of l-cysteine. Here, we examine the β-substitution capability of the OASS from Citrullus vulgaris (CvOASS), a putative l-mimosine synthase. While the previously reported mimosine synthase activity was not reproducible in our hands, we successfully identified non-native reactivity with a variety of O-nucleophiles. Optimization of reaction conditions for carboxylate and phenolate substrates led to distinct conditions that were leveraged for the preparative-scale synthesis of nsAAs. We further show this enzyme is capable of C-C bond formation through a β-alkylation reaction with an activated nitroalkane. To facilitate understanding of this enzyme, we determined the crystal structure of the enzyme bound to PLP as the internal aldimine at 1.55 Å, revealing key features of the active site and providing information that may guide subsequent development of CvOASS as a practical biocatalyst.

PubMed: 35476889
DOI: 10.1002/cbic.202200157

実験手法

X-RAY DIFFRACTION (1.55 Å)