5VRI

Crystal structure of SsoPox AsA6 mutant (F46L-C258A-W263M-I280T) - closed form

Summary for 5VRI

• Entry DOI: 10.2210/pdb5vri/pdb
• Related: 2VC5 2VC7 5VRK 5VSA 5W3U 5W3W 5W3Z
• Descriptor: Aryldialkylphosphatase, COBALT (II) ION, FE (II) ION, ... (6 entities in total)
• Functional Keywords: lactonase, phosphotriesterase, mutants, quorum sensing, organophosphate, organophosphorous, insecticides., hydrolase
• Biological source: Sulfolobus solfataricus
• Total number of polymer chains: 4
• Total formula weight: 143266.90

Authors

Hiblot, J.,Gotthard, G.,Jacquet, P.,Daude, D.,Bergonzi, C.,Chabriere, E.,Elias, M. (deposition date: 2017-05-10, release date: 2017-12-20, Last modification date: 2023-11-15)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (2.15 Å)