5W3U

Crystal structure of SsoPox AsB5 mutant (V27A-I76T-Y97W-Y99F-L130P-L226V)

5W3U の概要

• エントリーDOI: 10.2210/pdb5w3u/pdb
• 関連するPDBエントリー: 2vc5 2vc7 5VRI 5VRK 5VSA 5W3W 5W3Z
• 分子名称: Aryldialkylphosphatase, FE (II) ION, COBALT (II) ION, ... (6 entities in total)
• 機能のキーワード: lactonase, phosphotriesterase, mutants, quorum sensing, organophosphate, organophosphorous, insecticides., hydrolase
• 由来する生物種: Sulfolobus solfataricus
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 143360.71

構造登録者

Hiblot, J.,Gotthard, G.,Jacquet, P.,Daude, D.,Bergonzi, C.,Chabriere, E.,Elias, M. (登録日: 2017-06-08, 公開日: 2017-12-20, 最終更新日: 2021-08-18)

主引用文献

Jacquet, P.,Hiblot, J.,Daude, D.,Bergonzi, C.,Gotthard, G.,Armstrong, N.,Chabriere, E.,Elias, M.
Rational engineering of a native hyperthermostable lactonase into a broad spectrum phosphotriesterase.
Sci Rep, 7:16745-16745, 2017

PubMed Abstract: The redesign of enzyme active sites to alter their function or specificity is a difficult yet appealing challenge. Here we used a structure-based design approach to engineer the lactonase SsoPox from Sulfolobus solfataricus into a phosphotriesterase. The five best variants were characterized and their structure was solved. The most active variant, αsD6 (V27A-Y97W-L228M-W263M) demonstrates a large increase in catalytic efficiencies over the wild-type enzyme, with increases of 2,210-fold, 163-fold, 58-fold, 16-fold against methyl-parathion, malathion, ethyl-paraoxon, and methyl-paraoxon, respectively. Interestingly, the best mutants are also capable of degrading fensulfothion, which is reported to be an inhibitor for the wild-type enzyme, as well as others that are not substrates of the starting template or previously reported W263 mutants. The broad specificity of these engineered variants makes them promising candidates for the bioremediation of organophosphorus compounds. Analysis of their structures reveals that the increase in activity mainly occurs through the destabilization of the active site loop involved in substrate binding, and it has been observed that the level of disorder correlates with the width of the enzyme specificity spectrum. This finding supports the idea that active site conformational flexibility is essential to the acquisition of broader substrate specificity.

PubMed: 29196634
DOI: 10.1038/s41598-017-16841-0

実験手法

X-RAY DIFFRACTION (2.5 Å)