Summary for 8YYR
| Entry DOI | 10.2210/pdb8yyr/pdb |
| Descriptor | Putative ATP-dependent b-aminoacyl-ACP synthetase, [(2~{R},3~{S},4~{R},5~{R})-5-(6-aminopurin-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methyl ~{N}-[(3~{S})-3-azanyl-3-(2-bromophenyl)propanoyl]sulfamate (3 entities in total) |
| Functional Keywords | hitachimycin, polyketide biosynthesis, atp binding, adenylation, ligase |
| Biological source | Embleya scabrispora |
| Total number of polymer chains | 1 |
| Total formula weight | 60203.04 |
| Authors | Wang, D.,Miyanaga, A.,Chisuga, T.,Kudo, F.,Eguchi, T. (deposition date: 2024-04-04, release date: 2024-06-05, Last modification date: 2024-08-14) |
| Primary citation | Wang, D.,Miyanaga, A.,Chisuga, T.,Kudo, F.,Eguchi, T. Engineering the Substrate Specificity of (S)-beta-Phenylalanine Adenylation Enzyme HitB. Chembiochem, 25:e202400383-e202400383, 2024 Cited by PubMed Abstract: Adenylation enzymes catalyze the selective incorporation of aminoacyl building blocks in the biosynthesis of nonribosomal peptides and related natural products. Although β-amino acid units are one of the important aminoacyl building blocks in natural product biosynthesis, very little is known about the engineering of β-amino acid adenylation enzymes. In this study, we engineered the substrate specificity of the (S)-β-phenylalanine adenylation enzyme, HitB, involved in the biosynthesis of macrolactam polyketide hitachimycin. Based on the previously determined structure of HitB wild-type, we mutated Phe328 and Ser293, which are located near the meta and ortho position of the (S)-β-phenylalanine moiety, respectively. As a result, the HitB F328V and F328L mutants efficiently activated meta-substituted (S)-β-phenylalanine analogs, and the HitB T293G and T293S mutants efficiently activated ortho-substituted (S)-β-phenylalanine analogs. Structural analysis of the HitB F328L and T293G mutants with the corresponding nonhydrolyzable intermediate analogs revealed an enlarged substrate binding pocket for (S)-β-phenylalanine analogs, providing detailed insights into the structural basis for creating enzyme substrate promiscuity. Our findings may be useful for production of various β-amino acid-containing natural product analogs. PubMed: 38805007DOI: 10.1002/cbic.202400383 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.8 Å) |
Structure validation
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