PDBsum entry 5vrk

Hydrolase

PDB id: 5vrk

Contents

Protein chains

314 a.a.

Ligands

GOL ×7

EDO ×4

Metals

FE2 ×2

_CO ×2

Waters ×634

PDB id:

5vrk

Name:

Hydrolase

Title:

Crystal structure of ssopox asa6 mutant (f46l-c258a-w263m-i280t) - open form

Structure:

Aryldialkylphosphatase. Chain: a, b. Synonym: paraoxonase,ssopox,phosphotriesterase-like lactonase. Engineered: yes

Source:

Sulfolobus solfataricus. Organism_taxid: 2287. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.40Å R-factor: 0.141 R-free: 0.175

Authors:

J.Hiblot,G.Gotthard,P.Jacquet,D.Daude,C.Bergonzi,E.Chabriere,M.Elias

Key ref:

P.Jacquet et al. (2017). Rational engineering of a native hyperthermostable lactonase into a broad spectrum phosphotriesterase. Sci Rep, 7, 16745. PubMed id: 29196634

Date:

10-May-17 Release date: 10-Jan-18

Protein chains

Q97VT7  (PHP_SULSO) -  Aryldialkylphosphatase from Saccharolobus solfataricus (strain ATCC 35092 / DSM 1617 / JCM 11322 / P2)

Seq: 314 a.a.

Struc: 314 a.a.*

* PDB and UniProt seqs differ at 5 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.3.1.8.1 - aryldialkylphosphatase.
• Reaction: An aryl dialkyl phosphate + H2O = dialkyl phosphate + an aryl alcohol
• Cofactor: Divalent cation

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Added reference

Sci Rep 7:16745 (2017)

PubMed id: 29196634

Rational engineering of a native hyperthermostable lactonase into a broad spectrum phosphotriesterase.

P.Jacquet, J.Hiblot, D.Daudé, C.Bergonzi, G.Gotthard, N.Armstrong, E.Chabrière, M.Elias.

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.