par Pucci, Fabrizio 
;Kwasigroch, Jean-Marc ;Rooman, Marianne
Référence Methods in molecular biology, 2112, page (59-73)
Publication Publié, 2020-07-01
;Kwasigroch, Jean-Marc ;Rooman, Marianne Référence Methods in molecular biology, 2112, page (59-73)
Publication Publié, 2020-07-01
Article révisé par les pairs
| Résumé : | The rational design of enzymes is a challenging research field, which plays an important role in the optimization of a wide series of biotechnological processes. Computational approaches allow screening all possible amino acid substitutions in a target protein and to identify a subset likely to have the desired properties. They can thus be used to guide and restrict the huge, time-consuming search in sequence space to reach protein optimality. Here we present HoTMuSiC, a tool that predicts the impact of point mutations on the protein melting temperature, which uses the experimental or modeled protein structure as sole input and is available at the dezyme.com website. Its main advantages include accuracy and speed, which makes it a perfect instrument for thermal stability engineering projects aiming at designing new proteins that feature increased heat resistance or remain active and stable in nonphysiological conditions. We set up a HoTMuSiC-based pipeline, which uses additional information to avoid mutations of functionally important residues, identified as being too well conserved among homologous proteins or too close to annotated functional sites. The efficiency of this pipeline is successfully demonstrated on Rhizomucor miehei lipase. |
