5A25

Rational engineering of a mesophilic carbonic anhydrase to an extreme halotolerant biocatalyst

5A25 の概要

• エントリーDOI: 10.2210/pdb5a25/pdb
• 分子名称: CARBONIC ANHYDRASE 2, ZINC ION, GLYCEROL, ... (6 entities in total)
• 機能のキーワード: lyase, protein engineering, biocatalysis
• 由来する生物種: BOS TAURUS (CATTLE)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 59495.50

構造登録者

Warden, A.,Newman, J.,Peat, T.S.,Seabrook, S.,Williams, M.,Dojchinov, G.,Haritos, V. (登録日: 2015-05-12, 公開日: 2015-05-20, 最終更新日: 2024-01-10)

主引用文献

Warden, A.C.,Williams, M.,Peat, T.S.,Seabrook, S.A.,Newman, J.,Dojchinov, G.,Haritos, V.S.
Rational Engineering of a Mesohalophilic Carbonic Anhydrase to an Extreme Halotolerant Biocatalyst.
Nat.Commun., 6:10278-, 2015

PubMed Abstract: Enzymes expressed by highly salt-tolerant organisms show many modifications compared with salt-affected counterparts including biased amino acid and lower α-helix content, lower solvent accessibility and negative surface charge. Here, we show that halotolerance can be generated in an enzyme solely by modifying surface residues. Rational design of carbonic anhydrase II is undertaken in three stages replacing 18 residues in total, crystal structures confirm changes are confined to surface residues. Catalytic activities and thermal unfolding temperatures of the designed enzymes increase at high salt concentrations demonstrating their shift to halotolerance, whereas the opposite response is found in the wild-type enzyme. Molecular dynamics calculations reveal a key role for sodium ions in increasing halotolerant enzyme stability largely through interactions with the highly ordered first Na(+) hydration shell. For the first time, an approach to generate extreme halotolerance, a trait with broad application in industrial biocatalysis, in a wild-type enzyme is demonstrated.

PubMed: 26687908
DOI: 10.1038/NCOMMS10278

実験手法

X-RAY DIFFRACTION (1.9 Å)