7N4W
Complex structure of ROTU4 with rotundine
Summary for 7N4W
| Entry DOI | 10.2210/pdb7n4w/pdb |
| Related | 7N4Z 7N53 7N54 |
| Descriptor | ROTU4, SULFATE ION, GLYCEROL, ... (5 entities in total) |
| Functional Keywords | transcription factor, protein engineering, specificity, synthetic biology, transcription |
| Biological source | Salmonella enterica subsp. enterica serovar Typhimurium str. 14028S |
| Total number of polymer chains | 1 |
| Total formula weight | 22688.97 |
| Authors | |
| Primary citation | d'Oelsnitz, S.,Kim, W.,Burkholder, N.T.,Javanmardi, K.,Thyer, R.,Zhang, Y.,Alper, H.S.,Ellington, A.D. Using fungible biosensors to evolve improved alkaloid biosyntheses. Nat.Chem.Biol., 18:981-989, 2022 Cited by PubMed Abstract: A key bottleneck in the microbial production of therapeutic plant metabolites is identifying enzymes that can improve yield. The facile identification of genetically encoded biosensors can overcome this limitation and become part of a general method for engineering scaled production. We have developed a combined screening and selection approach that quickly refines the affinities and specificities of generalist transcription factors; using RamR as a starting point, we evolve highly specific (>100-fold preference) and sensitive (half-maximum effective concentration (EC) < 30 μM) biosensors for the alkaloids tetrahydropapaverine, papaverine, glaucine, rotundine and noscapine. High-resolution structures reveal multiple evolutionary avenues for the malleable effector-binding site and the creation of new pockets for different chemical moieties. These sensors further enabled the evolution of a streamlined pathway for tetrahydropapaverine, a precursor to four modern pharmaceuticals, collapsing multiple methylation steps into a single evolved enzyme. Our methods for evolving biosensors enable the rapid engineering of pathways for therapeutic alkaloids. PubMed: 35799063DOI: 10.1038/s41589-022-01072-w PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.64 Å) |
Structure validation
Download full validation report
