1C7I

THERMOPHYLIC PNB ESTERASE

「1C00」から置き換えられました

1C7I の概要

• エントリーDOI: 10.2210/pdb1c7i/pdb
• 分子名称: PROTEIN (PARA-NITROBENZYL ESTERASE), CALCIUM ION (3 entities in total)
• 機能のキーワード: alpha-beta hydrolase, pnb esterase, directed evolution, thermophile, hydrolase
• 由来する生物種: Bacillus subtilis
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 54263.94

構造登録者

Spiller, B.,Gershenson, A.,Arnold, F.,Stevens, R. (登録日: 2000-02-21, 公開日: 2000-03-29, 最終更新日: 2023-12-27)

主引用文献

Spiller, B.,Gershenson, A.,Arnold, F.H.,Stevens, R.C.
A structural view of evolutionary divergence.
Proc.Natl.Acad.Sci.USA, 96:12305-12310, 1999

PubMed Abstract: Two directed evolution experiments on p-nitrobenzyl esterase yielded one enzyme with a 100-fold increased activity in aqueous-organic solvents and another with a 17 degrees C increase in thermostability. Structures of the wild type and its organophilic and thermophilic counterparts are presented at resolutions of 1.5 A, 1.6 A, and 2.0 A, respectively. These structures identify groups of interacting mutations and demonstrate how directed evolution can traverse complex fitness landscapes. Early-generation mutations stabilize flexible loops not visible in the wild-type structure and set the stage for further beneficial mutations in later generations. The mutations exert their influence on the esterase structure over large distances, in a manner that would be difficult to predict. The loops with the largest structural changes generally are not the sites of mutations. Similarly, none of the seven amino acid substitutions in the organophile are in the active site, even though the enzyme experiences significant changes in the organization of this site. In addition to reduction of surface loop flexibility, thermostability in the evolved esterase results from altered core packing, helix stabilization, and the acquisition of surface salt bridges, in agreement with other comparative studies of mesophilic and thermophilic enzymes. Crystallographic analysis of the wild type and its evolved counterparts reveals networks of mutations that collectively reorganize the active site. Interestingly, the changes that led to diversity within the alpha/beta hydrolase enzyme family and the reorganization seen in this study result from main-chain movements.

PubMed: 10535917
DOI: 10.1073/pnas.96.22.12305

実験手法

X-RAY DIFFRACTION (2 Å)