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Literature summary 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)

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

Protein Variants

EC Number Protein Variants Comment Organism
3.7.1.9 D224A aggregates into inclusion bodies and thus can not be purified Pseudomonas fluorescens
3.7.1.9 H252A aggregates into inclusion bodies and thus can not be purified Pseudomonas fluorescens
3.7.1.9 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.7.1.9 0.00074
-
 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate
-
 Pseudomonas fluorescens
3.7.1.9 0.0073
-
 2-hydroxy-6-oxo-7-methylocta-2,4-dienoate 150 mM phosphate, 25°C, pH 7.5 Pseudomonas fluorescens
3.7.1.9 0.009
-
 2-hydroxy-6-oxohepta-2,4-dienoate
-
 Pseudomonas fluorescens
3.7.1.9 0.0095
-
 2-hydroxy-6-oxoocta-2,4-dienoate
-
 Pseudomonas fluorescens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.7.1.9 31489
-
 2 * 32000, SDS-PAGE, 2 * 31489, amino acid sequence Pseudomonas fluorescens
3.7.1.9 32000
-
 2 * 32000, SDS-PAGE, 2 * 31489, amino acid sequence Pseudomonas fluorescens
3.7.1.9 58000
-
 gel filtration Pseudomonas fluorescens

Organism

EC Number Organism UniProt Comment Textmining
3.7.1.9 Pseudomonas fluorescens
-
 IPO1
-
3.7.1.9 Pseudomonas fluorescens IPO1
-
 IPO1
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.7.1.9 6.1
-
 S103A mutant Pseudomonas fluorescens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.7.1.9 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate + H2O slight hydrolysis Pseudomonas fluorescens ?
-
 ?
3.7.1.9 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate + H2O slight hydrolysis Pseudomonas fluorescens IPO1 ?
-
 ?
3.7.1.9 2-hydroxy-6-oxo-7-methylocta-2,4-dienoate + H2O
-
 Pseudomonas fluorescens ?
-
 ?
3.7.1.9 2-hydroxy-6-oxo-7-methylocta-2,4-dienoate + H2O
-
 Pseudomonas fluorescens IPO1 ?
-
 ?
3.7.1.9 2-hydroxy-6-oxohepta-2,4-dienoate + H2O
-
 Pseudomonas fluorescens ?
-
 ?
3.7.1.9 2-hydroxy-6-oxohepta-2,4-dienoate + H2O
-
 Pseudomonas fluorescens IPO1 ?
-
 ?
3.7.1.9 2-hydroxy-6-oxoocta-2,4-dienoate + H2O
-
 Pseudomonas fluorescens ?
-
 ?
3.7.1.9 2-hydroxy-6-oxoocta-2,4-dienoate + H2O
-
 Pseudomonas fluorescens IPO1 ?
-
 ?

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
3.7.1.9 2-hydroxy-6-oxo-7-methylocta-2,4-dienoate hydrolase
-
 Pseudomonas fluorescens
3.7.1.9 CumD
-
 Pseudomonas fluorescens
3.7.1.9 meta-cleavage compound hydrolase
-
 Pseudomonas fluorescens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.7.1.9 47.5
-
-
 Pseudomonas fluorescens

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
3.7.1.9 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]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.7.1.9 4
-
 stable for 3 days Pseudomonas fluorescens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.7.1.9 0.03
-
 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate
-
 Pseudomonas fluorescens
3.7.1.9 18
-
 2-hydroxy-6-oxohepta-2,4-dienoate
-
 Pseudomonas fluorescens
3.7.1.9 21
-
 2-hydroxy-6-oxo-7-methylocta-2,4-dienoate 150 mM phosphate, 25°C, pH 7.5 Pseudomonas fluorescens
3.7.1.9 41
-
 2-hydroxy-6-oxoocta-2,4-dienoate
-
 Pseudomonas fluorescens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.7.1.9 8
-
-
 Pseudomonas fluorescens

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
3.7.1.9 7 9
-
 Pseudomonas fluorescens