Cloned (Comment) | Organism |
---|---|
gene VMUT_2255, phylogenetic analysis and sequence comparisons, recombinant enzyme expression in Escherichia coli strain BL21(DE3)-pGro7/GroEL | Vulcanisaeta moutnovskia |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme, with or without substrate, sitting drop vapor diffusion method, mixing of 20 mg/ml protein with different crystallization solutions, i.e. 400 mM ammonium dihydrogen phosphate plus G9 (100 mM sodium citrate, pH 5.6, 10 mM ferric chloride, and 10% v/v jeffamine M-600), C4 (100 mM sodium HEPES, pH 7.5, 800 mM sodium, and potassium dihydrogen phosphate) or E7 (100 mM Tris, pH 8.5, 1.5 M ammonium sulfate, and 12% v/v glycerol) in a 2:1 and 1:1 protein to reservoir ratio. The crystals from the G9 and E7 conditions have the same P64 space group and the ones issued from the C4 condition have an alternative P622 group space. The crystals from E7 are soaked into the well solution supplemented by 2 mM of substrate N-oxodecanoyl-DL-homoserine lactone for 1 min prior to flash cooling, crystal growth at °C, 2 weeks, X-ray diffraction structure determination and analysis at 1.8-2.35 A resolution, molecular replacement using the SsoPox structure as a starting template (PDB ID 2vc5), modelling | Vulcanisaeta moutnovskia |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | gamma- and delta-dodecanoic lactone substrates have an allosteric profile of hydrolysis | Vulcanisaeta moutnovskia | |
0.0021 | - |
dodecanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.047 | - |
nonanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.105 | - |
undecanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.12 | - |
undecanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.186 | - |
N-3-oxo-octanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.231 | - |
N-3-oxo-decanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.262 | - |
N-octanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.55 | - |
(R)-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.75 | - |
(S)-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.839 | - |
dodecanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.872 | - |
heptanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
1.22 | - |
nonanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
3.709 | - |
rac-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | required, VmoLac is a bi-cobalt metalloenzyme. The two metal cations are coordinated by four histidines, His23, His25, His171, and His200, an aspartic acid Asp257, and a carboxylated lysine residue Lys238 | Vulcanisaeta moutnovskia | |
additional information | the enzyme contains a bi-metallic active site that is coordinated by four histidines, an aspartic acid and a carboxylated lysine. The bi-metallic center acts as a Lewis acid and activates a bridging, putatively catalytic, water molecule into a hydroxide anion, which subsequently serves as a nucleophile for the organophosphorous compounds or lactone hydrolysis | Vulcanisaeta moutnovskia |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
an N-acyl-L-homoserine lactone + H2O | Vulcanisaeta moutnovskia | - |
an N-acyl-L-homoserine | - |
? | |
homo-L-cysteine thiolactone + H2O | Vulcanisaeta moutnovskia | - |
homo-L-cysteine | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Vulcanisaeta moutnovskia | F0QXN6 | a hyperthermophilic crenarchaeon | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-gamma-caprolactone + H2O | - |
Vulcanisaeta moutnovskia | 4-hydroxycaproate | - |
? | |
(S)-gamma-caprolactone + H2O | - |
Vulcanisaeta moutnovskia | caproate | - |
? | |
an N-acyl-L-homoserine lactone + H2O | - |
Vulcanisaeta moutnovskia | an N-acyl-L-homoserine | - |
? | |
dodecanoic-delta-lactone + H2O | - |
Vulcanisaeta moutnovskia | 5-hydroxydodecanoate | - |
? | |
dodecanoic-gamma-lactone + H2O | - |
Vulcanisaeta moutnovskia | dodecanoate | - |
? | |
epsilon-caprolactone + H2O | - |
Vulcanisaeta moutnovskia | 6-hydroxycaproate | - |
? | |
gamma-heptanolide + H2O | - |
Vulcanisaeta moutnovskia | ? | - |
? | |
heptanoic-gamma-lactone + H2O | - |
Vulcanisaeta moutnovskia | 4-hydroxyheptanoate | - |
? | |
homo-L-cysteine thiolactone + H2O | - |
Vulcanisaeta moutnovskia | homo-L-cysteine | - |
? | |
additional information | the enzyme is also active on diethyl-paraoxon, dimethyl-paraoxon, diethyl-parathion, dimethyl-parathion, and malathion. The lactone ring hydrolysis is followed by acidification of the medium for which Cresol purple serves as pH indicator. No or poor activity is detected against phenylacetate, dihydrocoumarin, 4-nitrophenyl acetate, 4-nitrophenyl decanoate, and AHLs with short aliphatic substituents (4 to 6 carbon atoms), e.g. N-hexanoyl-DL-homoserine lactone, N-butanoyl-DL-homoserine lactone, and N-3-oxo-hexanoyl-DL-homoserine lactone. AHLs with longer substituents are better substrates for VmoLac, including C8 and C10-AHLs. Oxo-lactones are the best substrates for VmoLac. gamma-caprolactone and gamma-heptalactone are degraded with approximately the same catalytic efficiencies, and oxo-lactones with long aliphatic chains such as nonanoic-gamma-lactone and undecanoic-delta-lactone, are hydrolyzed with high catalytic efficiencies | Vulcanisaeta moutnovskia | ? | - |
? | |
N-3-oxo-decanoyl-DL-homoserine lactone + H2O | - |
Vulcanisaeta moutnovskia | N-3-oxo-decanoyl-DL-homoserine | - |
? | |
N-3-oxo-octanoyl-DL-homoserine lactone + H2O | - |
Vulcanisaeta moutnovskia | N-3-oxo-octanoyl-DL-homoserine | - |
? | |
N-octanoyl-DL-homoserine lactone + H2O | - |
Vulcanisaeta moutnovskia | N-octanoyl-DL-homoserine | - |
? | |
nonanoic-delta-lactone + H2O | - |
Vulcanisaeta moutnovskia | 5-hydroxynonanoate | - |
? | |
nonanoic-delta-lactone + H2O | - |
Vulcanisaeta moutnovskia | 5-hydroxynonanoic acid | - |
? | |
nonanoic-gamma-lactone + H2O | best substrate | Vulcanisaeta moutnovskia | 4-hydroxynonanoate | - |
? | |
nonanoic-gamma-lactone + H2O | best substrate | Vulcanisaeta moutnovskia | 4-hydroxynonanoic acid | - |
? | |
rac-gamma-caprolactone + H2O | - |
Vulcanisaeta moutnovskia | 4-hydroxycaproate | - |
? | |
undecanoic-delta-lactone + H2O | - |
Vulcanisaeta moutnovskia | 5-hydroxyundecanoate | - |
? | |
undecanoic-gamma-lactone + H2O | - |
Vulcanisaeta moutnovskia | 4-hydroxyundecanoate | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | VmoLac is roughly globular and exhibits a (beta/alpha)8 barrel topology | Vulcanisaeta moutnovskia |
Synonyms | Comment | Organism |
---|---|---|
More | cf. EC 3.1.8.1 | Vulcanisaeta moutnovskia |
quorum quenching lactonase | - |
Vulcanisaeta moutnovskia |
VmoLac | - |
Vulcanisaeta moutnovskia |
VMUT_2255 | - |
Vulcanisaeta moutnovskia |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
40 | - |
assay at | Vulcanisaeta moutnovskia |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.16 | - |
dodecanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.35 | - |
N-3-oxo-octanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.47 | - |
N-3-oxo-decanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
0.59 | - |
N-octanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
1.89 | - |
(S)-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
3.04 | - |
(R)-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
8.86 | - |
undecanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
27.25 | - |
heptanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
28.08 | - |
dodecanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
44.49 | - |
nonanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
60.78 | - |
undecanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
88.91 | - |
nonanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
112.3 | - |
rac-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.3 | - |
assay at | Vulcanisaeta moutnovskia |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the phosphotriesterase-like lactonases (PLLs) family of enzyme. Phosphotriesterase-like lactonases (PLLs) compose a class of lactonases that has long been mistaken for the organophosphorus-degrading enzymes phosphotriesterases (PTEs, EC 3.1.8.1). Several PLLs have initially been isolated and characterized by virtue of their organophosphorous insecticide degrading abilities. PLLs belong to the amidohydrolase superfamily and exhibit a (beta/alpha)8 topology containing a bi-metallic active site that is coordinated by four histidines, an aspartic acid and a carboxylated lysine. PTEs and PLLs mainly differ by the size and conformations of the active site loops 7 and 8 that mediate substrate specificity | Vulcanisaeta moutnovskia |
additional information | the active site loops 7 and 8 mediate the substrate specificity. VmoLac possesses a bi-cobalt active site. Active site structure and structure comparisons, overview | Vulcanisaeta moutnovskia |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.88 | - |
N-3-oxo-octanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
2.04 | - |
N-3-oxo-decanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
2.25 | - |
N-octanoyl-DL-homoserine lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
2.52 | - |
(S)-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
5.53 | - |
(R)-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
30.28 | - |
rac-gamma-caprolactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
31.25 | - |
heptanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
33.47 | - |
dodecanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
72.88 | - |
nonanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
73.83 | - |
undecanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
76.19 | - |
dodecanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
578.86 | - |
undecanoic-delta-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia | |
946.6 | - |
nonanoic-gamma-lactone | pH 8.0, 25°C, recombinant enzyme | Vulcanisaeta moutnovskia |