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Literature summary extracted from

  • Janzen, E.; Mueller, M.; Kolter-Jung, D.; Kneen, M.M.; McLeish, M.J.; Pohl, M.
    Characterization of benzaldehyde lyase from Pseudomonas fluorescens: A versatile enzyme for asymmetric C-C bond formation (2006), Bioorg. Chem., 34, 345-361.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.2.38 expressed in Escherichia coli strain SG13009 Pseudomonas fluorescens

Protein Variants

EC Number Protein Variants Comment Organism
4.1.2.38 A28S mutant shows decarboxylase activity towards 2-keto acids Pseudomonas fluorescens
4.1.2.38 A480I reduced lyase activity Pseudomonas fluorescens
4.1.2.38 F484I reduced lyase activity Pseudomonas fluorescens
4.1.2.38 H286A reduced lyase activity Pseudomonas fluorescens

Inhibitors

EC Number Inhibitors Comment Organism Structure
4.1.2.38 Mg2+ inhibitory above 7.5 mM Pseudomonas fluorescens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.2.38 0.045
-
 Benzoin mutant enzyme A480I, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 0.047
-
 Benzoin mutant enzyme A28S, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 0.048
-
 Benzoin wild type enzyme, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 0.055
-
 Benzoin mutant enzyme H286A, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 0.067
-
 Benzoin mutant enzyme F484I, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 0.3
-
 2-hydroxypropiophenone wild type enzyme, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 0.6
-
 2-hydroxypropiophenone mutant enzyme A28S, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 0.9
-
 2-hydroxypropiophenone mutant enzyme H286A, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 1.2
-
 2-hydroxypropiophenone mutant enzyme F484I, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 5
-
 2-hydroxypropiophenone mutant enzyme A480I, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 10
-
 benzaldehyde in 50 mM potassium phosphate at 30°C and pH 7.5 Pseudomonas fluorescens

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.1.2.38 Mg2+ dependent, 1-5 mM Pseudomonas fluorescens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
4.1.2.38 54000
-
 4 * 54000, gel filtration Pseudomonas fluorescens
4.1.2.38 59800
-
 His-tagged BAL subunit, SDS-PAGE and MALDI-TOF-MS Pseudomonas fluorescens
4.1.2.38 216000
-
 gel filtration Pseudomonas fluorescens

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
4.1.2.38 DMSO the addition of 20% (v/v) DMSO is useful to enhance both the solubility of aromatic substrates and products and the stability of BAL Pseudomonas fluorescens

Organism

EC Number Organism UniProt Comment Textmining
4.1.2.38 Pseudomonas fluorescens
-
 strain biovar I
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.2.38 immobilized metal ion chromatography, after which the enzyme shows only 35-45% of its activity, however, activity is completely regained after incubation in the presence of Mg2+ and thiamine diphosphate Pseudomonas fluorescens

Storage Stability

EC Number Storage Stability Organism
4.1.2.38 30°C, in cofactor-free potassium phosphate buffer at pH 7.0, 3 h, 65% loss of activity Pseudomonas fluorescens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.2.38 2-hydroxypropiophenone
-
 Pseudomonas fluorescens ?
-
 ?
4.1.2.38 benzaldehyde + (R)-2,2-dimethyl-1,3-dioxolan-4-carbaldehyde
-
 Pseudomonas fluorescens acyloin + benzoin
-
 ?
4.1.2.38 benzoin
-
 Pseudomonas fluorescens benzaldehyde + benzaldehyde
-
 r
4.1.2.38 additional information unable to cleave acetoin Pseudomonas fluorescens ?
-
 ?

Subunits

EC Number Subunits Comment Organism
4.1.2.38 tetramer 4 * 54000, gel filtration Pseudomonas fluorescens

Synonyms

EC Number Synonyms Comment Organism
4.1.2.38 benzaldehyde lyase the wild type enzyme does not show any decarboxylase activity towards 2-keto acids Pseudomonas fluorescens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.1.2.38 35
-
 maximal activity for the benzoin lyase reaction Pseudomonas fluorescens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.1.2.38 25
-
 a concentration of 1 mM MgSO4 and 0.01 mM thiamine diphosphate is sufficient to keep BAL stable at 25°C Pseudomonas fluorescens
4.1.2.38 37
-
 reaction temperature shall not exceed 37°C, in the presence of 5 mM MgSO4 and 0.1 mM thiamine diphosphate the enzyme's activity is reduced by only 15% over 3 h at 37°C, however, a concentration of 1 mM MgSO4 and 0.01 mM thiamine diphosphate leads to more than 40% inactivation within 3 h at 37°C Pseudomonas fluorescens

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.2.38 1.6
-
 2-hydroxypropiophenone mutant enzyme A28S, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 2.8
-
 2-hydroxypropiophenone mutant enzyme H286A, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 3.4
-
 2-hydroxypropiophenone wild type enzyme, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 4
-
 2-hydroxypropiophenone mutant enzyme F484I, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 4.9
-
 2-hydroxypropiophenone mutant enzyme A480I, at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 6.6
-
 Benzoin mutant enzyme A480I, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 9.5
-
 Benzoin mutant enzyme A28S, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 22
-
 Benzoin mutant enzyme F484I, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 56.4
-
 Benzoin mutant enzyme H286A, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens
4.1.2.38 66.9
-
 Benzoin wild type enzyme, in the presence of 15% (v/v) PEG-400, in 50 mM Tris at 30°C and pH 7.5 Pseudomonas fluorescens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.1.2.38 8
-
 pH-optimum of both the lyase and the ligase reaction, in potassium phosphate, Tris, and imidazole buffers Pseudomonas fluorescens

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
4.1.2.38 6 8 in potassium phosphate, Tris, and imidazole buffers, pH values below 6 lead to complete inactivation after about one day, while the enzyme shows some activity when kept above pH 8 Pseudomonas fluorescens

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.2.38 thiamine diphosphate dependent, the optimal concentration is 0.1-5 mM, a significant loss of activity is observed with a concentration below 0.01 mM Pseudomonas fluorescens

pI Value

EC Number Organism Comment pI Value Maximum pI Value
4.1.2.38 Pseudomonas fluorescens isoelectric focusing
-
 4.6