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Literature summary extracted from

  • Park, Y.J.; Yoon, S.J.; Lee, H.B.
    A novel dienelactone hydrolase from the thermoacidophilic archaeon Sulfolobus solfataricus P1: purification, characterization, and expression (2010), Biochim. Biophys. Acta, 1800, 1164-1172.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.45 gene dlh, expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Saccharolobus solfataricus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.45 C151S site-directed mutagenesis, almost inactive mutant Saccharolobus solfataricus
3.1.1.45 D198N site-directed mutagenesis, almost inactive mutant Saccharolobus solfataricus
3.1.1.45 H229N site-directed mutagenesis, almost inactive mutant Saccharolobus solfataricus
3.1.1.45 S102A site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme Saccharolobus solfataricus
3.1.1.45 S103A site-directed mutagenesis, 20% reduced activity compared to the wild-type enzyme Saccharolobus solfataricus
3.1.1.45 S120A site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme Saccharolobus solfataricus
3.1.1.45 S139A site-directed mutagenesis, the mutant is 3-5fold less active than the wild-type enzyme Saccharolobus solfataricus
3.1.1.45 S147A site-directed mutagenesis, 20% reduced activity compared to the wild-type enzyme Saccharolobus solfataricus
3.1.1.45 S210A site-directed mutagenesis, the mutant shows similar activity like the wild-type enzyme Saccharolobus solfataricus

General Stability

EC Number General Stability Organism
3.1.1.45 the purified enzyme shows high stability against denaturing agents, including various detergents, urea, and organic solvents Saccharolobus solfataricus

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.1.1.45 4-chloromercuribenzoate 78% inhibition at 5 mM Saccharolobus solfataricus
3.1.1.45 diethyl dicarbonate 71% inhibition at 5 mM Saccharolobus solfataricus
3.1.1.45 diisopropylfluorophosphate complete inhibition at 0.01 mM Saccharolobus solfataricus
3.1.1.45 HgCl2 92% inhibition at 5 mM Saccharolobus solfataricus
3.1.1.45 additional information no or poor inhibition by eserine, pyridoxal 5'-phosphate, and phenylglyoxal Saccharolobus solfataricus
3.1.1.45 paraoxon complete inhibition at 5 mM Saccharolobus solfataricus
3.1.1.45 PMSF complete inhibition at 0.01 mM Saccharolobus solfataricus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.45 additional information
-
additional information kinetics with other 4-nitrophenyl esters for carboxylesterase activity, overview Saccharolobus solfataricus
3.1.1.45 0.1017
-
trans-dienelactone pH 5.0, 60°C Saccharolobus solfataricus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.1.45 29000
-
SDS-PAGE, gel filtration and MALDI TOF mass spectrometry, recombinant enzyme Saccharolobus solfataricus

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.1.1.45 dienelactone + H2O Saccharolobus solfataricus
-
maleylacetate
-
?
3.1.1.45 dienelactone + H2O Saccharolobus solfataricus P1
-
maleylacetate
-
?
3.1.1.45 additional information Saccharolobus solfataricus the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family and containing a catalytic triad composed of Cys151, Asp198, and His229 in the active site ?
-
?
3.1.1.45 additional information Saccharolobus solfataricus P1 the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family and containing a catalytic triad composed of Cys151, Asp198, and His229 in the active site ?
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.45 Saccharolobus solfataricus B9WPN8 gene dlh; DSM1616
-
3.1.1.45 Saccharolobus solfataricus P1 B9WPN8 gene dlh; DSM1616
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.1.45 recombinant enzyme 150fold from Escherichia coli strain BL21(DE3) by anion exchange and hydrophobic interaction chromatography, dialysis, and another anion exchange chromatography Saccharolobus solfataricus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.1.45 990
-
wild-type enzyme, substrate trans-dienelactone, pH 5.0, 60°C Saccharolobus solfataricus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.45 dienelactone + H2O
-
Saccharolobus solfataricus maleylacetate
-
?
3.1.1.45 dienelactone + H2O
-
Saccharolobus solfataricus P1 maleylacetate
-
?
3.1.1.45 additional information the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family and containing a catalytic triad composed of Cys151, Asp198, and His229 in the active site Saccharolobus solfataricus ?
-
?
3.1.1.45 additional information the enzyme also shows carboxylesterase activity toward 4-nitrophenyl esters ranging from butyrate to laurate Saccharolobus solfataricus ?
-
?
3.1.1.45 additional information the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family and containing a catalytic triad composed of Cys151, Asp198, and His229 in the active site Saccharolobus solfataricus P1 ?
-
?
3.1.1.45 additional information the enzyme also shows carboxylesterase activity toward 4-nitrophenyl esters ranging from butyrate to laurate Saccharolobus solfataricus P1 ?
-
?
3.1.1.45 trans-dienelactone + H2O the enzyme displays substrate specificities toward trans-dienelactone, not cis-isomer Saccharolobus solfataricus maleylacetate
-
?
3.1.1.45 trans-dienelactone + H2O the enzyme displays substrate specificities toward trans-dienelactone, not cis-isomer Saccharolobus solfataricus P1 maleylacetate
-
?

Subunits

EC Number Subunits Comment Organism
3.1.1.45 monomer 1 * 29000, SDS-PAGE, gel filtration and MALDI TOF mass spectrometry, recombinant enzyme Saccharolobus solfataricus

Synonyms

EC Number Synonyms Comment Organism
3.1.1.45 dienelactone hydrolase
-
Saccharolobus solfataricus
3.1.1.45 More the enzyme is a typical dienelactone hydrolase belonging to alpha/beta hydrolase family Saccharolobus solfataricus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.45 74
-
-
Saccharolobus solfataricus

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.1.1.45 30 90 assay range Saccharolobus solfataricus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.45 90
-
purified enzyme, 50% activity remaining after 50 h Saccharolobus solfataricus

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.1.1.45 9409
-
trans-dienelactone pH 5.0, 60°C Saccharolobus solfataricus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.45 5
-
-
Saccharolobus solfataricus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.1.45 3 9 assay range Saccharolobus solfataricus

General Information

EC Number General Information Comment Organism
3.1.1.45 physiological function the hydrolysis of dienelactone plays a key role for the microbial degradation of chloroaromatics via chlorocatechols Saccharolobus solfataricus

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.1.1.45 0.0925
-
trans-dienelactone pH 5.0, 60°C Saccharolobus solfataricus