4UHE
Structural studies of a thermophilic esterase from Thermogutta terrifontis (malate bound)
Summary for 4UHE
| Entry DOI | 10.2210/pdb4uhe/pdb |
| Related | 4UHC 4UHD 4UHF 4UHH |
| Descriptor | ESTERASE, TRIETHYLENE GLYCOL, CHLORIDE ION, ... (5 entities in total) |
| Functional Keywords | hydrolase, alpha beta hydrolase |
| Biological source | PLANCTOMYCETES BACTERIUM R1 |
| Total number of polymer chains | 1 |
| Total formula weight | 31528.07 |
| Authors | Sayer, C.,Isupov, M.N.,Bonch-Osmolovskaya, E.,Littlechild, J.A. (deposition date: 2015-03-24, release date: 2015-06-10, Last modification date: 2024-01-10) |
| Primary citation | Sayer, C.,Isupov, M.N.,Bonch-Osmolovskaya, E.,Littlechild, J.A. Structural Studies of a Thermophilic Esterase from a New Planctomycetes Species, Thermogutta Terrifontis. FEBS J., 282:2846-, 2015 Cited by PubMed Abstract: Thermogutta terrifontis esterase (TtEst), a carboxyl esterase identified in the novel thermophilic bacterium T. terrifontis from the phylum Planctomycetes, has been cloned and over-expressed in Escherichia coli. The enzyme has been characterized biochemically and shown to have activity towards small p-nitrophenyl (pNP) carboxylic esters, with optimal activity for pNP-propionate. The enzyme retained 95% activity after incubation for 1 h at 80 °C. The crystal structures of the native TtEst and its complexes with the substrate analogue D-malate and the product acetate have been determined to high resolution. The bound ligands have allowed the identification of the carboxyl and alcohol binding pockets in the enzyme active site. Comparison of TtEst with structurally related enzymes provides insight into how differences in their catalytic activity can be rationalized based upon the properties of the amino acid residues in their active site pockets. The mutant enzymes L37A and L251A have been constructed to extend the substrate range of TtEst towards the larger butyrate and valerate pNP-esters. These mutant enzymes have also shown a significant increase in activity towards acetate and propionate pNP esters. A crystal structure of the L37A mutant was determined with the butyrate product bound in the carboxyl pocket of the active site. The mutant structure shows an expansion of the pocket that binds the substrate carboxyl group, which is consistent with the observed increase in activity towards pNP-butyrate. PubMed: 26011036DOI: 10.1111/FEBS.13326 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.16 Å) |
Structure validation
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