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Information on EC 3.1.1.81 - quorum-quenching N-acyl-homoserine lactonase and Organism(s) Saccharolobus solfataricus and UniProt Accession Q97VT7

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EC Tree
     3 Hydrolases
         3.1 Acting on ester bonds
             3.1.1 Carboxylic-ester hydrolases
                3.1.1.81 quorum-quenching N-acyl-homoserine lactonase
IUBMB Comments
Acyl-homoserine lactones (AHLs) are produced by a number of bacterial species and are used by them to regulate the expression of virulence genes in a process known as quorum-sensing. Each bacterial cell has a basal level of AHL and, once the population density reaches a critical level, it triggers AHL-signalling which, in turn, initiates the expression of particular virulence genes . Plants or animals capable of degrading AHLs would have a therapeutic advantage in avoiding bacterial infection as they could prevent AHL-signalling and the expression of virulence genes in quorum-sensing bacteria . N-(3-Oxohexanoyl)-L-homoserine lactone, N-(3-oxododecanoyl)-L-homoserine lactone, N-butanoyl-L-homoserine lactone and N-(3-oxooctanoyl)-L-homoserine lactone can act as substrates .
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Saccharolobus solfataricus
UNIPROT: Q97VT7
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Word Map
The taxonomic range for the selected organisms is: Saccharolobus solfataricus
The expected taxonomic range for this enzyme is: Bacteria, Archaea, Eukaryota
Synonyms
ahl-lactonase, ahl-acylase, ahl-degrading enzyme, aiia lactonase, aii20j, n-acyl-homoserine lactonase, n-acylhomoserine lactonase, n-acyl homoserine lactonase, quorum-quenching enzyme, ahl-1, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
lactonase/phosphotriesterase
-
N-acyl homoserine lactonase
-
phosphotriesterase-like lactonase
-
SsoPox
quorum-sensing enzyme
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
N-acyl-L-homoserine-lactone lactonohydrolase
Acyl-homoserine lactones (AHLs) are produced by a number of bacterial species and are used by them to regulate the expression of virulence genes in a process known as quorum-sensing. Each bacterial cell has a basal level of AHL and, once the population density reaches a critical level, it triggers AHL-signalling which, in turn, initiates the expression of particular virulence genes [5]. Plants or animals capable of degrading AHLs would have a therapeutic advantage in avoiding bacterial infection as they could prevent AHL-signalling and the expression of virulence genes in quorum-sensing bacteria [5]. N-(3-Oxohexanoyl)-L-homoserine lactone, N-(3-oxododecanoyl)-L-homoserine lactone, N-butanoyl-L-homoserine lactone and N-(3-oxooctanoyl)-L-homoserine lactone can act as substrates [5].
CAS REGISTRY NUMBER
COMMENTARY hide
389867-43-0
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
5-thiobutyl butyrolactone + H2O
?
show the reaction diagram
TBBL
-
-
?
an N-acyl-L-homoserine lactone + H2O
an N-acyl-L-homoserine
show the reaction diagram
-
-
-
?
N-(3-oxodecanoyl)-L-homoserine lactone + H2O
N-(3-oxodecanoyl)-L-homoserine
show the reaction diagram
the enzyme has a preference for N-acyl homoserine lactones with acyl chain lengths of at least 8 carbon atoms
-
-
?
N-(3-oxododecanoyl)-L-homoserine lactone + H2O
N-(3-oxododecanoyl)-L-homoserine
show the reaction diagram
the enzyme has a preference for N-acyl homoserine lactones with acyl chain lengths of at least 8 carbon atoms
-
-
?
N-(3-oxohexanoyl)-L-homoserine lactone + H2O
N-(3-oxohexanoyl)-L-homoserine
show the reaction diagram
the enzyme has a preference for N-acyl homoserine lactones with acyl chain lengths of at least 8 carbon atoms
-
-
?
N-(3-oxooctanoyl)-L-homoserine lactone + H2O
N-(3-oxooctanoyl)-L-homoserine
show the reaction diagram
the enzyme has a preference for N-acyl homoserine lactones with acyl chain lengths of at least 8 carbon atoms
-
-
?
N-3-oxododecanoyl-L-homoserine lactone + H2O
N-3-oxododecanoyl-L-homoserine
show the reaction diagram
-
-
-
?
undecanoic-delta-lactone + H2O
5-hydroxyundecanoic acid
show the reaction diagram
-
-
-
?
undecanoic-gamma-lactone + H2O
4-hydroxyundecanoic acid
show the reaction diagram
-
-
-
?
an N-acyl-L-homoserine lactone + H2O
an N-acyl-L-homoserine
show the reaction diagram
-
-
-
-
?
ethyl paraoxon + H2O
4-nitrophenol + diethyl phosphate
show the reaction diagram
-
-
-
-
?
N-(3-oxododecanoyl)-L-homoserine lactone + H2O
N-(3-oxododecanoyl)-L-homoserine
show the reaction diagram
-
-
-
-
?
N-dodecanoyl-L-homoserine lactone + H2O
N-dodecanoyl-L-homoserine
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
an N-acyl-L-homoserine lactone + H2O
an N-acyl-L-homoserine
show the reaction diagram
-
-
-
?
an N-acyl-L-homoserine lactone + H2O
an N-acyl-L-homoserine
show the reaction diagram
-
-
-
-
?
additional information
?
-
the phosphotriesterase-like lactonase enzyme is bifunctional showing lactonase (EC 3.1.1.81) and phosphotriesterase (EC 3.1.8.1) activities
-
-
?
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
petroleum ether
-
petroleum ether slightly reduces the activity down to 78%
-
Toluene
-
toluene slightly reduced the activity down to 74%
xylene
-
xylene decreases the enzyme activity by more than 75%
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Methylcyclohexane
-
about 250% activity at 100% (v/v)
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.05
N-(3-oxodecanoyl)-L-homoserine lactone
pH 8.0, 25°C
0.17
N-(3-oxododecanoyl)-L-homoserine lactone
pH 8.0, 25°C
5.6
N-(3-oxohexanoyl)-L-homoserine lactone
pH 8.0, 25°C
0.16
N-(3-oxooctanoyl)-L-homoserine lactone
pH 8.0, 25°C
additional information
additional information
kinetic analysis of lactonase activity
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.5
N-(3-oxodecanoyl)-L-homoserine lactone
pH 8.0, 25°C
0.95
N-(3-oxododecanoyl)-L-homoserine lactone
pH 8.0, 25°C
0.52
N-(3-oxohexanoyl)-L-homoserine lactone
pH 8.0, 25°C
1
N-(3-oxooctanoyl)-L-homoserine lactone
pH 8.0, 25°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
290
N-(3-oxodecanoyl)-L-homoserine lactone
pH 8.0, 25°C
5.5
N-(3-oxododecanoyl)-L-homoserine lactone
pH 8.0, 25°C
0.093
N-(3-oxohexanoyl)-L-homoserine lactone
pH 8.0, 25°C
6.6
N-(3-oxooctanoyl)-L-homoserine lactone
pH 8.0, 25°C
0.141 - 31.6
N-3-oxododecanoyl-L-homoserine lactone
2.55 - 9650
undecanoic-delta-lactone
0.905 - 573
undecanoic-gamma-lactone
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
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)
metabolism
-
the enzyme also exhibits promiscuous phosphotriesterase activity for the degradation of organophosphorous chemicals including insecticides and chemical warfare agents
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
34000
x * 34000, SDS-PAGE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
x * 34000, SDS-PAGE
?
-
x * 35000, SDS-PAGE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
hanging drop vapor diffusion method, using 20-30% (w/v) PEG 8000 and 50 mM Tris-HCl buffer (pH 8.0)
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C258L/I261F/W263A
F46L/C258A/W263M/I280T
the mutant exhibits enhanced phosphotriesterase activity compared to the wild type enzyme
L72I/Y99F/I122L/L228M/F229S/W263L
the mutant exhibits enhanced phosphotriesterase activity compared to the wild type enzyme
V27A/I76T/Y97W/Y99F/L130P/L226V
the mutant exhibits enhanced phosphotriesterase activity compared to the wild type enzyme
V27A/Y97W/L228M/W263M
the mutant demonstrates a large increase in catalytic efficiencies over the wild-type enzyme, with increases of 2210fold, 163fold, 58fold, 16fold against methyl parathion, malathion, ethyl paraoxon, and methyl paraoxon, respectively
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
W263I
a variant with increased catalytic effectiveness against N-3-oxododecanoyl-L-homoserine lactone
W263I
-
the mutant has increased lactonase and phosphotriesterase activities compared to the wild type
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
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
106°C is the melting temperature. The temperatures required to lose one half of enzymatic activity in 5 minutes are 92°C and 130°C in liquid and solid states respectively. More than 30% activity remains after autoclaving the enzyme powder at 121°C for 15 min
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
when the enzyme is immobilized on nanoalumina membranes 25% of the activity is retained upon immobilization
with the liquid enzyme, the remaining activity decreases down to 40% at the highest dose of beta-radiation (100 kGy)
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
the enzyme activity is not affected by acetone, acetonitrile, butanone, butyl acetate, chloroform, dichloromethane, diethyl ether, ethanol, ethyl acetate, isopropanol, methanol, and methoxypropanol
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation and Superdex 75 gel filtration
Co2+ IMAC column chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) cells
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)
overexpressed in Pseudomonas putida KT2440
expressed in Escherichia coli BL21(DE3) cells
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
lactonase SsoPox-W263I is significantly more effective than the tested quorum sensing inhibitors at their respective concentration optimum in inhibition of the virulence of 51 clinical Pseudomonas aeruginosa isolates from diabetic foot ulcers. SsoPox may be incorporated into medical devices such as functionalised catheters and antivirulence dressings
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ng, F.S.; Wright, D.M.; Seah, S.Y.
Characterization of a phosphotriesterase-like lactonase from Sulfolobus solfataricus and its immobilization for disruption of quorum sensing
Appl. Environ. Microbiol.
77
1181-1186
2010
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus, Saccharolobus solfataricus P2 (Q97VT7)
Manually annotated by BRENDA team
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
Biotechnol. Bioeng.
113
724-734
2016
Saccharolobus solfataricus (Q97VT7)
Manually annotated by BRENDA team
Guendouze, A.; Plener, L.; Bzdrenga, J.; Jacquet, P.; Remy, B.; Elias, M.; Lavigne, J.P.; Daude, D.; Chabriere, E.
Effect of quorum quenching lactonase in clinical isolates of Pseudomonas aeruginosa and comparison with quorum sensing inhibitors
Front. Microbiol.
8
227
2017
Saccharolobus solfataricus (Q97VT7), Saccharolobus solfataricus DSM 1617 (Q97VT7)
Manually annotated by BRENDA team
Rhoads, M.K.; Hauk, P.; Gupta, V.; Bookstaver, M.L.; Stephens, K.; Payne, G.F.; Bentley, W.E.
Modification and assembly of a versatile lactonase for bacterial quorum quenching
Molecules
23
E341
2018
Saccharolobus solfataricus
Manually annotated by BRENDA team
Remy, B.; Plener, L.; Poirier, L.; Elias, M.; Daude, D.; Chabriere, E.
Harnessing hyperthermostable lactonase from Sulfolobus solfataricus for biotechnological applications
Sci. Rep.
6
37780
2016
Saccharolobus solfataricus
Manually annotated by BRENDA team
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
2017
Saccharolobus solfataricus (Q97VT7)
Manually annotated by BRENDA team