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Literature summary for 3.1.1.81 extracted from

  • Del Giudice, I.; Coppolecchia, R.; Merone, L.; Porzio, E.; Carusone, T.M.; Mandrich, L.; Worek, F.; Manco, G.
    An efficient thermostable organophosphate hydrolase and its application in pesticide decontamination (2016), Biotechnol. Bioeng., 113, 724-734 .
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene ssopox and gene ssopox-pte, sequence comparisons, recombinant expression of wild-type, point mutation, and chimeric mutant enzymes in Escherichia coli strains TOP10 and BL21(DE3) Saccharolobus solfataricus

Protein Variants

Protein Variants Comment Organism
C258L/I261F/W263A site-directed mutagenesis, the mutant is not able to hydrolyze C8-HSL or C10-HSL and its paraoxonase activity is 3fold higher than the lactonase activity on 5-thiobutyl butyrolactone, as opposed to the wild-type paraoxonase activity which is 760fold lower than the lactonase activity. The combination of C258L, I261F, and W263A mutations in the SsoPox triple mutant improves the hydrolytic specific activity in terms of kcat/KM toward paraoxon 12fold, the kcat 294fold compared to wild-type SsoPox, while the KM value increases Saccharolobus solfataricus
additional information evolution of a lactonase into a phosphotriesterase, semi-rational engineering approach is used to design an efficient and thermostable organophosphate hydrolase, starting from enzyme SsoPox from Sulfolobus solfataricus as a lactonase scaffold. In particular, by in vitro evolution of the SsoPox ancillary promiscuous activity, the triple mutant C258L/I261F/W263A is obtained which, retaining its inherent stability, shows an enhancement of its hydrolytic activity on paraoxon up to 300fold. The mutant is tested in formulations of different solvents (methanol or ethanol) or detergents (SDS or a commercial soap) for the cleaning of pesticide-contaminated surfaces. Construction of a chimeric gene ssopox-pte by insertion of 16 conserved residues of pte gene in the ssopox sequence. Recombination by DNA StEP between ssopox-pte chimera and ssopox gene Saccharolobus solfataricus
W263F site-directed mutagenesis, the W263 residue is previously demonstrated to be involved in the formation of an hydrophobic channel for the substrate leaving group, the mutant enzyme shows decreased lactonase activity compared to the wild-type Saccharolobus solfataricus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetic analysis of lactonase activity Saccharolobus solfataricus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ activates, required Saccharolobus solfataricus
additional information the enzyme has a binuclear metal-centre. The activity depends on the presence of divalent metal cations, the highest activity is observed with Co2+. The binuclear centre is used to activate a bridging water molecule to a hydroxide ion and the substrate for nucleophilic attack by polarizing the phosphoryl-oxygen bond. The nucleophilic bridging hydroxide ion attacks the electrophilic centre (phosphorus or carbon) via a SN2 mechanism, forming transition states that bridge the two metals Saccharolobus solfataricus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
an N-acyl-L-homoserine lactone + H2O Saccharolobus solfataricus
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an N-acyl-L-homoserine
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?
additional information Saccharolobus solfataricus the phosphotriesterase-like lactonase enzyme is bifunctional showing lactonase (EC 3.1.1.81) and phosphotriesterase (EC 3.1.8.1) activities ?
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?

Organism

Organism UniProt Comment Textmining
Saccharolobus solfataricus Q97VT7 i.e. Sulfolobus solfataricus
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Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5-thiobutyl butyrolactone + H2O TBBL Saccharolobus solfataricus ?
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?
an N-acyl-L-homoserine lactone + H2O
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Saccharolobus solfataricus an N-acyl-L-homoserine
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?
additional information the phosphotriesterase-like lactonase enzyme is bifunctional showing lactonase (EC 3.1.1.81) and phosphotriesterase (EC 3.1.8.1) activities Saccharolobus solfataricus ?
-
?
additional information the enzyme has activity on acyl-homoserine lactones and 5-thiobutyl butyrolactone (TBBL). Substrate docking analysis. The C258 residue in the active site is involved in an interaction with the oxygen atom from the amide in the lactone analogue. C258 residue might have a key role in the lactone hydrolysis mechanism Saccharolobus solfataricus ?
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?

Synonyms

Synonyms Comment Organism
lactonase/phosphotriesterase
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Saccharolobus solfataricus
phosphotriesterase-like lactonase
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Saccharolobus solfataricus
PHP
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Saccharolobus solfataricus
PLL
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Saccharolobus solfataricus
SsoPox
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Saccharolobus solfataricus

General Information

General Information Comment Organism
evolution enzyme SsoPox belongs to the phosphotriesterase-like lactonase (PLL) family of enzymes, to the PLL-A subfamily, a group of lactonases showing a preference for acyl-homoserine lactones. SsoPox shares only about 30% sequence identity with phosphotriesterases (PTEs) but all amino acids coordinating the binuclear metal-centre are conserved. The coexistence of lactonase and phosphotriesterase activities has been already reported for many members of PLL family Saccharolobus solfataricus
physiological function SsoPox is a thermostable phosphotriesterase-like lactonase (PLL) that hydrolyses lactones (primary activity) and, at a lower rate, neurotoxic organophosphorus compounds (promiscuous activity) Saccharolobus solfataricus