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Literature summary extracted from

  • Franceschini, S.; van Beek, H.L.; Pennetta, A.; Martinoli, C.; Fraaije, M.W.; Mattevi, A.
    Exploring the structural basis of substrate preferences in Baeyer-Villiger monooxygenases: insight from steroid monooxygenase (2012), J. Biol. Chem., 287, 22626-22634.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.14.13.54 expressed in Escherichia coli BL21(DE3) CodonPlus cells Rhodococcus rhodochrous

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.13.54 sitting drop vapor diffusion method, using 1.8-2.2 M MgSO4, 0.1 M MES/HCl, pH 6.2-6.5 Rhodococcus rhodochrous

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.54 K295A the mutant is 3times more active than the wild type enzyme Rhodococcus rhodochrous
1.14.13.54 L500Y the mutant is about 2.5times more active than the wild type enzyme Rhodococcus rhodochrous
1.14.13.54 P157Q the mutant shows wild type activity Rhodococcus rhodochrous
1.14.13.54 T345L the mutation turns the enzyme inactive against progesterone without altering the catalytic efficiency for phenylacetone Rhodococcus rhodochrous
1.14.13.54 V291A the mutant is less active than the wild type enzyme Rhodococcus rhodochrous

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.54 0.2
-
 phenylacetone mutant enzyme L500Y, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 0.2
-
 phenylacetone mutant enzyme V291A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 0.6
-
 phenylacetone mutant enzyme P157Q, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 0.8
-
 phenylacetone mutant enzyme K295A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1
-
 phenylacetone wild type enzyme, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 2.6
-
 phenylacetone mutant enzyme T345L, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.13.54 progesterone + NADPH + H+ + O2 Rhodococcus rhodochrous
-
 Testosterone acetate + NADP+ + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.54 Rhodococcus rhodochrous O50641
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.13.54 Ni2+-affinity column chromatography and Superdex 75 gel filtration Rhodococcus rhodochrous

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.54 phenylacetone + NADPH + H+ + O2
-
 Rhodococcus rhodochrous benzylacetate + NADP+ + H2O
-
 ?
1.14.13.54 progesterone + NADPH + H+ + O2
-
 Rhodococcus rhodochrous Testosterone acetate + NADP+ + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
1.14.13.54 Baeyer-Villiger monooxygenase
-
 Rhodococcus rhodochrous
1.14.13.54 Steroid monooxygenase
-
 Rhodococcus rhodochrous
1.14.13.54 STMO
-
 Rhodococcus rhodochrous

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.54 0.2
-
 phenylacetone mutant enzyme T345L, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 0.6
-
 phenylacetone mutant enzyme P157Q, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1
-
 phenylacetone mutant enzyme L500Y, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1.5
-
 phenylacetone wild type enzyme, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1.8
-
 phenylacetone mutant enzyme V291A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 3
-
 phenylacetone mutant enzyme K295A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.13.54 FAD
-
 Rhodococcus rhodochrous
1.14.13.54 NADPH
-
 Rhodococcus rhodochrous

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.54 0.1
-
 progesterone kcat_Km less than 0.1 1/sec*min, mutant enzyme L500Y, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1.2
-
 phenylacetone mutant enzyme K295A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1.2
-
 phenylacetone mutant enzyme T345L, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1.4
-
 progesterone mutant enzyme K295A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1.5
-
 phenylacetone wild type enzyme, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1.7
-
 progesterone mutant enzyme V291A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 1.8
-
 progesterone wild type enzyme, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 2.1
-
 phenylacetone mutant enzyme L500Y, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 2.2
-
 phenylacetone mutant enzyme V291A, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 2.6
-
 phenylacetone mutant enzyme P157Q, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 3.9
-
 progesterone mutant enzyme P157Q, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous
1.14.13.54 5.2
-
 progesterone mutant enzyme T345L, in 50 mM Tris/HCl, pH 7.5, temperature not specified in the publication Rhodococcus rhodochrous