1T4M
STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION
Summary for 1T4M
| Entry DOI | 10.2210/pdb1t4m/pdb |
| Related | 1T2N |
| Descriptor | LIPASE A, POTASSIUM ION (3 entities in total) |
| Functional Keywords | alpha/beta hydrolase, hydrolase |
| Biological source | Bacillus subtilis |
| Cellular location | Secreted: P37957 |
| Total number of polymer chains | 1 |
| Total formula weight | 19515.02 |
| Authors | Rajakumara, E.,Sankaranarayanan, R. (deposition date: 2004-04-30, release date: 2004-11-23, Last modification date: 2023-08-23) |
| Primary citation | Acharya, P.,Rajakumara, E.,Sankaranarayanan, R.,Rao, N.M. Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase J.Mol.Biol., 341:1271-1281, 2004 Cited by PubMed Abstract: Variation in gene sequences generated by directed evolution approaches often does not assure a minimalist design for obtaining a desired property in proteins. While screening for enhanced thermostability, structural information was utilized in selecting mutations that are generated by error-prone PCR. By this approach we have increased the half-life of denaturation by 300-fold compared to the wild-type Bacillus subtilis lipase through three point mutations generated by only two cycles of error-prone PCR. At lower temperatures the activity parameters of the thermostable mutants are unaltered. High-resolution crystal structures of the mutants show subtle changes, which include stacking of tyrosine residues, peptide plane flipping and a better anchoring of the terminus, that challenge rational design and explain the structural basis for enhanced thermostability. The approach may offer an efficient and minimalist solution for the enhancement of a desired property of a protein. PubMed: 15321721DOI: 10.1016/j.jmb.2004.06.059 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2 Å) |
Structure validation
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