1t4m
From Proteopedia
|
STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION
Overview
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.
About this Structure
1T4M is a Single protein structure of sequence from Bacillus subtilis with K as ligand. Active as Triacylglycerol lipase, with EC number 3.1.1.3 Full crystallographic information is available from OCA.
Reference
Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase., Acharya P, Rajakumara E, Sankaranarayanan R, Rao NM, J Mol Biol. 2004 Aug 27;341(5):1271-81. PMID:15321721
Page seeded by OCA on Wed Nov 21 02:59:16 2007
