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Literature summary extracted from

  • Kadow, M.; Loschinski, K.; Sass, S.; Schmidt, M.; Bornscheuer, U.
    Completing the series of BVMOs involved in camphor metabolism of Pseudomonas putida NCIMB 10007 by identification of the two missing genes, their functional expression in E. coli, and biochemical characterization (2012), Appl. Microbiol. Biotechnol., 96, 419-429.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.160 expression in Escherichia coli Pseudomonas putida
1.14.14.155 expressed in Escherichia coli BL21(DE3) cells Pseudomonas putida

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.160 0.028
-
 (+-)-cis-bicyclo[3.2.0]hept-2-en-6-one pH 9.0, 20°C Pseudomonas putida
1.14.13.160 0.127
-
 Cyclobutanone pH 9.0, 20°C Pseudomonas putida

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.13.160 60000
-
 x * 60000, SDS-PAGE, x * 63500, calculated, both for His-tagged protein Pseudomonas putida
1.14.13.160 63500
-
 x * 60000, SDS-PAGE, x * 63500, calculated, both for His-tagged protein Pseudomonas putida
1.14.14.155 40000
-
 x * 40000, SDS-PAGE Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.160 Pseudomonas putida H3JQW0
-
-
1.14.13.160 Pseudomonas putida NCIMB 10007 H3JQW0
-
-
1.14.14.155 Pseudomonas putida D7UER1
-
-
1.14.14.155 Pseudomonas putida NCIMB 10007 D7UER1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.160 recombinant enzyme Pseudomonas putida
1.14.14.155 HisTrap column chromatography Pseudomonas putida

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.14.13.160 2200
-
 pH 9.0, 20°C Pseudomonas putida
1.14.14.155 0.09
-
 crude extract, at pH 7.5 and 25°C Pseudomonas putida
1.14.14.155 0.46
-
 purified enzyme, at pH 7.5 and 25°C Pseudomonas putida

Storage Stability

EC Number Storage Stability Organism
1.14.13.160 4°C, 24 h, 50% loss of activity Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.160 (+-)-cis-bicyclo[3.2.0]hept-2-en-6-one + O2 + NADPH + H+
-
 Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.13.160 (+-)-cis-bicyclo[3.2.0]hept-2-en-6-one + O2 + NADPH + H+
-
 Pseudomonas putida NCIMB 10007 ? + NADP+ + H2O
-
 ?
1.14.13.160 2,3,4,5-tetramethyl-2-cyclopenten-1-one + O2 + NADPH + H+ 34% conversion Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.13.160 2,3,4,5-tetramethyl-2-cyclopenten-1-one + O2 + NADPH + H+ 34% conversion Pseudomonas putida NCIMB 10007 ? + NADP+ + H2O
-
 ?
1.14.13.160 2-cyclohexen-1-one + O2 + NADPH + H+ 74% conversion Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.13.160 2-cyclohexen-1-one + O2 + NADPH + H+ 74% conversion Pseudomonas putida NCIMB 10007 ? + NADP+ + H2O
-
 ?
1.14.13.160 2-cyclopenten-1-one + O2 + NADPH + H+ 62% conversion Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.13.160 3,5,5-trimethyl-2-cyclohexen-1-one + O2 + NADPH + H+ 22% conversion Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.13.160 3,5,5-trimethyl-2-cyclohexen-1-one + O2 + NADPH + H+ 22% conversion Pseudomonas putida NCIMB 10007 ? + NADP+ + H2O
-
 ?
1.14.13.160 3-methyl-2-cyclohexen-1-one + O2 + NADPH + H+ 13% conversion Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.13.160 3-methyl-2-cyclohexen-1-one + O2 + NADPH + H+ 13% conversion Pseudomonas putida NCIMB 10007 ? + NADP+ + H2O
-
 ?
1.14.13.160 3-methyl-2-cyclopenten-1-one + O2 + NADPH + H+ 13% conversion Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.13.160 cyclobutanone + O2 + NADPH + H+
-
 Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.13.160 norcamphor + O2 + NADPH + H+
-
 Pseudomonas putida ? + NADP+ + H2O
-
 ?
1.14.14.155 (+)-camphor + FMNH2 + O2 88% conversion Pseudomonas putida ? + FMN + H2O
-
 ?
1.14.14.155 (+)-camphor + FMNH2 + O2 88% conversion Pseudomonas putida NCIMB 10007 ? + FMN + H2O
-
 ?
1.14.14.155 (+/-)-cis-bicyclo[3.2.0]hept-2-en-6-one + FMNH2 + O2 99% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 (+/-)-cis-bicyclo[3.2.0]hept-2-en-6-one + FMNH2 + O2 99% conversion Pseudomonas putida NCIMB 10007 ?
-
 ?
1.14.14.155 (-)-camphor + FMNH2 + O2 91% conversion Pseudomonas putida ? + FMN + H2O
-
 ?
1.14.14.155 (-)-camphor + FMNH2 + O2 91% conversion Pseudomonas putida NCIMB 10007 ? + FMN + H2O
-
 ?
1.14.14.155 (R,R)-bicyclo[2.2.1]heptane-2,5-dione + FMNH2 + O2 26% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 (R,R)-bicyclo[2.2.1]heptane-2,5-dione + FMNH2 + O2 26% conversion Pseudomonas putida NCIMB 10007 ?
-
 ?
1.14.14.155 2,3,4,5-tetramethyl-2-cyclopenten-1-one + FMNH2 + O2 43% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 2,3,4,5-tetramethyl-2-cyclopenten-1-one + FMNH2 + O2 43% conversion Pseudomonas putida NCIMB 10007 ?
-
 ?
1.14.14.155 2-cyclohexen-1-one + FMNH2 + O2 50% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 2-cyclopenten-1-one + FMNH2 + O2 48% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 2-decanone + FMNH2 + O2 11% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 3,5,5-trimethyl-2-cyclohexen-1-one + FMNH2 + O2 21% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 3-methyl-2-cyclohexen-1-one + FMNH2 + O2 20% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 3-methyl-2-cyclopenten-1-one + FMNH2 + O2 10% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 4-phenyl-2-butanone + FMNH2 + O2 48% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 acetophenone + FMNH2 + O2 80% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 cyclobutanone + FMNH2 + O2 13% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 cyclohexanone + FMNH2 + O2 3% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 cyclopentanone + FMNH2 + O2 24% conversion Pseudomonas putida ?
-
 ?
1.14.14.155 norcamphor + FMNH2 + O2 77% conversion Pseudomonas putida ?
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.13.160 ? x * 60000, SDS-PAGE, x * 63500, calculated, both for His-tagged protein Pseudomonas putida
1.14.14.155 ? x * 40000, SDS-PAGE Pseudomonas putida

Synonyms

EC Number Synonyms Comment Organism
1.14.14.155 3,6-diketocamphane 1,6-monooxygenase
-
 Pseudomonas putida
1.14.14.155 3,6-DKCMO
-
 Pseudomonas putida

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.13.160 23
-
-
 Pseudomonas putida

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.14.13.160 20
-
 half-life 196 min Pseudomonas putida
1.14.13.160 25
-
 half-life 43 min Pseudomonas putida
1.14.13.160 30
-
 half-life 9 min Pseudomonas putida

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.14.13.160 9
-
-
 Pseudomonas putida

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.160 FAD
-
 Pseudomonas putida
1.14.14.155 FMN dependent on Pseudomonas putida
1.14.14.155 additional information dependent on Pseudomonas putida

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.160 14.7
-
 Cyclobutanone pH 9.0, 20°C Pseudomonas putida
1.14.13.160 49.3
-
 (+-)-cis-bicyclo[3.2.0]hept-2-en-6-one pH 9.0, 20°C Pseudomonas putida