8F7F

The condensation domain of surfactin A synthetase C in space group P43212

Summary for 8F7F

• Entry DOI: 10.2210/pdb8f7f/pdb
• Descriptor: Surfactin synthetase, GLYCEROL (3 entities in total)
• Functional Keywords: nrps, c domain, srfa-c, biosynthetic protein
• Biological source: Bacillus subtilis
• Total number of polymer chains: 1
• Total formula weight: 53947.14

Authors

Frota, N.,Pistofidis, A.,Folger, I.B.,Hilvert, D.,Schmeing, T.M. (deposition date: 2022-11-18, release date: 2023-11-22, Last modification date: 2024-06-12)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.62 Å)