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Literature summary for 3.7.1.9 extracted from

  • Saku, T.; Fushinobu, S.; Jun, J.S.; Ikeda, N.; Nojiri, H.; Yamane, H.; Omori, T.; Wakagi, T.
    Purification, characterization, and steady-state kinetics of a meta-cleavage compound hydrolase from Pseudomonas fluorescens IPO1 (2002), J. Biosci. Bioeng., 93, 568-574.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
into pUC119 and overexpression in Escherichia coli Pseudomonas fluorescens

Protein Variants

Protein Variants Comment Organism
D224A aggregates into inclusion bodies and thus can not be purified Pseudomonas fluorescens
H252A aggregates into inclusion bodies and thus can not be purified Pseudomonas fluorescens
S103A has 600000fold lower activity than that of the wild-type enzyme, can be found in the soluble fraction and thus can be purified Pseudomonas fluorescens

Inhibitors

Inhibitors Comment Organism Structure
PMSF 0.01 mM strongly inhibits, CumD activity being undetectable Pseudomonas fluorescens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00074
-
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate
-
Pseudomonas fluorescens
0.0073
-
2-hydroxy-6-oxo-7-methylocta-2,4-dienoate 150 mM phosphate, 25°C, pH 7.5 Pseudomonas fluorescens
0.009
-
2-hydroxy-6-oxohepta-2,4-dienoate
-
Pseudomonas fluorescens
0.0095
-
2-hydroxy-6-oxoocta-2,4-dienoate
-
Pseudomonas fluorescens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
31489
-
2 * 32000, SDS-PAGE, 2 * 31489, amino acid sequence Pseudomonas fluorescens
32000
-
2 * 32000, SDS-PAGE, 2 * 31489, amino acid sequence Pseudomonas fluorescens
58000
-
gel filtration Pseudomonas fluorescens

Organism

Organism UniProt Comment Textmining
Pseudomonas fluorescens
-
IPO1
-
Pseudomonas fluorescens IPO1
-
IPO1
-

Purification (Commentary)

Purification (Comment) Organism
to homogeneity, gel filtration Pseudomonas fluorescens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.1
-
S103A mutant Pseudomonas fluorescens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate + H2O slight hydrolysis Pseudomonas fluorescens ?
-
?
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate + H2O slight hydrolysis Pseudomonas fluorescens IPO1 ?
-
?
2-hydroxy-6-oxo-7-methylocta-2,4-dienoate + H2O
-
Pseudomonas fluorescens ?
-
?
2-hydroxy-6-oxo-7-methylocta-2,4-dienoate + H2O
-
Pseudomonas fluorescens IPO1 ?
-
?
2-hydroxy-6-oxohepta-2,4-dienoate + H2O
-
Pseudomonas fluorescens ?
-
?
2-hydroxy-6-oxohepta-2,4-dienoate + H2O
-
Pseudomonas fluorescens IPO1 ?
-
?
2-hydroxy-6-oxoocta-2,4-dienoate + H2O
-
Pseudomonas fluorescens ?
-
?
2-hydroxy-6-oxoocta-2,4-dienoate + H2O
-
Pseudomonas fluorescens IPO1 ?
-
?

Subunits

Subunits Comment Organism
dimer 2 * 32000, SDS-PAGE, 2 * 31489, amino acid sequence Pseudomonas fluorescens

Synonyms

Synonyms Comment Organism
2-hydroxy-6-oxo-7-methylocta-2,4-dienoate hydrolase
-
Pseudomonas fluorescens
CumD
-
Pseudomonas fluorescens
meta-cleavage compound hydrolase
-
Pseudomonas fluorescens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
47.5
-
-
Pseudomonas fluorescens

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
5 60 full enzyme activity up to 20°C for 30 min, and ca. 80% between 30 and 50°C, almost completely inactivated above 60°C Pseudomonas fluorescens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4
-
stable for 3 days Pseudomonas fluorescens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.03
-
2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate
-
Pseudomonas fluorescens
18
-
2-hydroxy-6-oxohepta-2,4-dienoate
-
Pseudomonas fluorescens
21
-
2-hydroxy-6-oxo-7-methylocta-2,4-dienoate 150 mM phosphate, 25°C, pH 7.5 Pseudomonas fluorescens
41
-
2-hydroxy-6-oxoocta-2,4-dienoate
-
Pseudomonas fluorescens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
-
Pseudomonas fluorescens

pH Stability

pH Stability pH Stability Maximum Comment Organism
7 9
-
Pseudomonas fluorescens