8F7G

The condensation domain of surfactin A synthetase C in space group P212121

8F7G の概要

• エントリーDOI: 10.2210/pdb8f7g/pdb
• 分子名称: Surfactin synthetase, GLYCEROL (3 entities in total)
• 機能のキーワード: nrps, c domain, srfa-c, biosynthetic protein
• 由来する生物種: Bacillus subtilis
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 53855.05

構造登録者

Frota, N.F.,Pistofidis, A.,Folger, I.B.,Hilvert, D.,Schmeing, T.M. (登録日: 2022-11-18, 公開日: 2023-11-22, 最終更新日: 2024-06-12)

主引用文献

Folger, I.B.,Frota, N.F.,Pistofidis, A.,Niquille, D.L.,Hansen, D.A.,Schmeing, T.M.,Hilvert, D.
High-throughput reprogramming of an NRPS condensation domain.
Nat.Chem.Biol., 20:761-769, 2024

PubMed Abstract: Engineered biosynthetic assembly lines could revolutionize the sustainable production of bioactive natural product analogs. Although yeast display is a proven, powerful tool for altering the substrate specificity of gatekeeper adenylation domains in nonribosomal peptide synthetases (NRPSs), comparable strategies for other components of these megaenzymes have not been described. Here we report a high-throughput approach for engineering condensation (C) domains responsible for peptide elongation. We show that a 120-kDa NRPS module, displayed in functional form on yeast, can productively interact with an upstream module, provided in solution, to produce amide products tethered to the yeast surface. Using this system to screen a large C-domain library, we reprogrammed a surfactin synthetase module to accept a fatty acid donor, increasing catalytic efficiency for this noncanonical substrate >40-fold. Because C domains can function as selectivity filters in NRPSs, this methodology should facilitate the precision engineering of these molecular assembly lines.

PubMed: 38308044
DOI: 10.1038/s41589-023-01532-x

実験手法

X-RAY DIFFRACTION (2.4 Å)