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Literature summary for 3.1.1.45 extracted from

  • Camara, B.; Marin, M.; Schloemann, M.; Hecht, H.J.; Junca, H.; Pieper, D.H.
    trans-Dienelactone hydrolase from Pseudomonas reinekei MT1, a novel zinc-dependent hydrolase (2008), Biochem. Biophys. Res. Commun., 376, 423-428.
    View publication on PubMed
Search for more literature for 3.1.1.45

Cloned(Commentary)

Cloned (Comment) Organism
into pGEM-T Easy and subsequently into pASK-IBA7plus for expression of the protein in Escherichia coli XL10-GOLD cells Pseudomonas reinekei

Protein Variants

Protein Variants Comment Organism
additional information to evaluate the metal dependence of trans-DLH, a strep-tagged protein is purified Pseudomonas reinekei

Inhibitors

Inhibitors Comment Organism Structure
EDTA
-
 Pseudomonas reinekei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.55
-
 trans-dienelactone
-
 Pseudomonas reinekei

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+
-
 Pseudomonas reinekei
Ni2+
-
 Pseudomonas reinekei
Zn2+
-
 Pseudomonas reinekei

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36940
-
 calculated molecular weight Pseudomonas reinekei
37000
-
 determined by SDS-PAGE Pseudomonas reinekei

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4-chloromuconolactone + H2O Pseudomonas reinekei
-
 maleylacetate + HCl
-
 ?
trans-dienelactone + H2O Pseudomonas reinekei
-
 maleylacetate
-
 ?

Organism

Organism UniProt Comment Textmining
Pseudomonas reinekei
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant and wild-type trans-DLH are purified from cell extracts of Escherichia coli XL10-GOLD and Pseudomonas reinekei by subsequent anionic exchange, hydrophobic interaction, gel filtration and anionic exchange chromatography, the strep-tagged recombinant protein is purified using a Strep-Tactin Sepharose column Pseudomonas reinekei

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
29
-
 strep-tagged protein, contains 1.9 mol of Zn2+, less than 0.1 mol of Mn2+, and 1.1 mol Ni2+ per mol of subunit Pseudomonas reinekei
34
-
 recombinant protein, contains 2.3 mol Zn2+, 0.8 mol Mn2+, and less than 0.1 mol Ni2+ per mol of subunit Pseudomonas reinekei
65
-
 wild-type, contains 2.4 mol Zn2+, 2.0 mol Mn2+, and 1.1 mol Ni2+ per mol of subunit Pseudomonas reinekei

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-chloromuconolactone + H2O
-
 Pseudomonas reinekei maleylacetate + HCl
-
 ?
trans-dienelactone + H2O
-
 Pseudomonas reinekei maleylacetate
-
 ?

Synonyms

Synonyms Comment Organism
trans-dienelactone hydrolase
-
 Pseudomonas reinekei
trans-DLH
-
 Pseudomonas reinekei

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
20
-
 standard condition Pseudomonas reinekei
55
-
 assay for determination of temperature-dependence of the activity Pseudomonas reinekei

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
15100
-
 trans-dienelactone recombinant protein Pseudomonas reinekei

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
 assay for determination of pH-dependence of the activity Pseudomonas reinekei
7.5
-
 activity assay Pseudomonas reinekei