Acharya_2004_J.Mol.Biol_341_1271

Reference

Title : Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase - Acharya_2004_J.Mol.Biol_341_1271
Author(s) : Acharya P , Rajakumara E , Sankaranarayanan R , Rao NM
Ref : Journal of Molecular Biology , 341 :1271 , 2004
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.

PubMedSearch : Acharya_2004_J.Mol.Biol_341_1271
PubMedID: 15321721
Gene_locus related to this paper: bacsu-lip

Related information

Gene_locus bacsu-lip
Family Lipase_2
Structure 3D2A    3D2B    3D2C    1T2N    1T4M

Citations formats

Acharya P, Rajakumara E, Sankaranarayanan R, Rao NM (2004)
Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase
Journal of Molecular Biology 341 :1271

Acharya P, Rajakumara E, Sankaranarayanan R, Rao NM (2004)
Journal of Molecular Biology 341 :1271