1T4M

STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION

1T4M の概要

• エントリーDOI: 10.2210/pdb1t4m/pdb
• 関連するPDBエントリー: 1T2N
• 分子名称: LIPASE A, POTASSIUM ION (3 entities in total)
• 機能のキーワード: alpha/beta hydrolase, hydrolase
• 由来する生物種: Bacillus subtilis
• 細胞内の位置: Secreted: P37957
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 19515.02

構造登録者

Rajakumara, E.,Sankaranarayanan, R. (登録日: 2004-04-30, 公開日: 2004-11-23, 最終更新日: 2023-08-23)

主引用文献

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

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: 15321721
DOI: 10.1016/j.jmb.2004.06.059

実験手法

X-RAY DIFFRACTION (2 Å)