6IYL
The structure of EntE with 3-cyanobenzoyl adenylate analog
Summary for 6IYL
| Entry DOI | 10.2210/pdb6iyl/pdb |
| Descriptor | 2,3-dihydroxybenzoate-AMP ligase component of enterobactin synthase multienzyme complex, 5'-O-[(3-cyanobenzene-1-carbonyl)sulfamoyl]adenosine (3 entities in total) |
| Functional Keywords | ligase, adenylation, non-ribosomal peptide biosynthesis |
| Biological source | Escherichia coli 1303 |
| Total number of polymer chains | 2 |
| Total formula weight | 123617.32 |
| Authors | Miyanaga, A.,Ishikawa, F. (deposition date: 2018-12-17, release date: 2019-04-17, Last modification date: 2023-11-22) |
| Primary citation | Ishikawa, F.,Miyanaga, A.,Kitayama, H.,Nakamura, S.,Nakanishi, I.,Kudo, F.,Eguchi, T.,Tanabe, G. An Engineered Aryl Acid Adenylation Domain with an Enlarged Substrate Binding Pocket. Angew.Chem.Int.Ed.Engl., 58:6906-6910, 2019 Cited by PubMed Abstract: Adenylation (A) domains act as the gatekeepers of non-ribosomal peptide synthetases (NRPSs), ensuring the activation and thioesterification of the correct amino acid/aryl acid building blocks. Aryl acid building blocks are most commonly observed in iron-chelating siderophores, but are not limited to them. Very little is known about the reprogramming of aryl acid A-domains. We show that a single asparagine-to-glycine mutation in an aryl acid A-domain leads to an enzyme that tolerates a wide range of non-native aryl acids. The engineered catalyst is capable of activating non-native aryl acids functionalized with nitro, cyano, bromo, and iodo groups, even though no enzymatic activity of wild-type enzyme was observed toward these substrates. Co-crystal structures with non-hydrolysable aryl-AMP analogues revealed the origins of this expansion of substrate promiscuity, highlighting an enlargement of the substrate binding pocket of the enzyme. Our findings may be exploited to produce diversified aryl acid containing natural products and serve as a template for further directed evolution in combinatorial biosynthesis. PubMed: 30945421DOI: 10.1002/anie.201900318 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.56 Å) |
Structure validation
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