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Literature summary for 3.5.5.1 extracted from

  • Zhang, Z.J.; Cai, R.F.; Xu, J.H.
    Characterization of a new nitrilase from Hoeflea phototrophica DFL-43 for a two-step one-pot synthesis of (S)-?-amino acids (2018), Appl. Microbiol. Biotechnol., 102, 6047-6056 .
    View publication on PubMed

Application

Application Comment Organism
synthesis a cascade reaction for the synthesis of optically pure (S)-beta-phenylalanine from benzoylacetonitrile was developed by coupling HpN with an omega-transaminase from Polaromonas sp. JS666 in toluene-water biphasic reaction system using beta-alanine as an amino donor. Various (S)-beta-amino acids can be produced from benzoylacetonitrile derivatives with moderate to high conversions (73-99%) and excellent enantioselectivity (above 99% enantiomeric exess). Great potential of this cascade reaction for the practical synthesis of (S)-beta-phenylalanine Hoeflea phototrophica

Cloned(Commentary)

Cloned (Comment) Organism
overexpressed in Escherichia coli BL21 (DE3) Hoeflea phototrophica

Inhibitors

Inhibitors Comment Organism Structure
Ag+ 10 mM, complete loss of activity Hoeflea phototrophica
Cu2+ 10 mM, 94% loss of activity Hoeflea phototrophica
EDTA 10 mM, 20% loss of activity Hoeflea phototrophica
Fe2+ 10 mM, 59% loss of activity Hoeflea phototrophica
Hg2+ 10 mM, complete loss of activity Hoeflea phototrophica
Mg2+ 10 mM, 15% loss of activity Hoeflea phototrophica
Mn2+ 10 mM, 24% loss of activity Hoeflea phototrophica
Ni2+ 10 mM, 83% loss of activity Hoeflea phototrophica
Pb2+ 10 mM, 15% loss of activity Hoeflea phototrophica
Zn2+ 10 mM, 20% loss of activity Hoeflea phototrophica

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2
-
benzoylacetonitrile pH 8.5, 50°C Hoeflea phototrophica

Organic Solvent Stability

Organic Solvent Comment Organism
Acetone 5% (v/v), about 60% loss of activity Hoeflea phototrophica
acetonitrile 5% (v/v), no loss of activity Hoeflea phototrophica
dimethylformamide 5% (v/v), about 15% loss of activity Hoeflea phototrophica
DMSO 5% (v/v), about 10% loss of activity Hoeflea phototrophica
Ethanol 5% (v/v), about 15% loss of activity Hoeflea phototrophica
Methanol 5% (v/v), no loss of activity Hoeflea phototrophica

Organism

Organism UniProt Comment Textmining
Hoeflea phototrophica
-
-
-
Hoeflea phototrophica DFL-43
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Hoeflea phototrophica

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-chloromandelonitrile + H2O 7% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica ?
-
?
2-chloromandelonitrile + H2O 7% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica DFL-43 ?
-
?
2-chlorophenylacetonitrile + H2O 38% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica ?
-
?
3-chlorobenzoylacetonitrile + H2O 43% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica ?
-
?
3-methoxylbenzoylacetonitrile + H2O 38% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica ?
-
?
4-chlorobenzoylacetonitrile + H2O 16% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica ?
-
?
acetonitrile + H2O 12% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica acetic acid + NH3
-
?
acetonitrile + H2O 12% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica DFL-43 acetic acid + NH3
-
?
benzoylacetonitrile + H2O
-
Hoeflea phototrophica ?
-
?
glycolonitrile + H2O 18% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica ?
-
?
mandelonitrile + H2O 32% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica ?
-
?
mandelonitrile + H2O 32% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica DFL-43 ?
-
?
additional information no activity detected with: benzonitrile, 3-cyanopyridine, 2,3-dicyanopyrazine Hoeflea phototrophica ?
-
-
additional information no activity detected with: benzonitrile, 3-cyanopyridine, 2,3-dicyanopyrazine Hoeflea phototrophica DFL-43 ?
-
-
phenylacetonitrile + H2O 160% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica ?
-
?
phenylacetonitrile + H2O 160% of the activity as compared to benzoylacetonitrile Hoeflea phototrophica DFL-43 ?
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
50
-
-
Hoeflea phototrophica

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
40 55 40°C: about 60% of maximal activity, 55°C: about 35% of maximal activity Hoeflea phototrophica

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
30
-
24 h, no loss of activity Hoeflea phototrophica
40
-
half-life: 12 h Hoeflea phototrophica
50
-
half-life: 3 h Hoeflea phototrophica
60
-
half-life: 1 h Hoeflea phototrophica

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
170
-
benzoylacetonitrile pH 8.5, 50°C Hoeflea phototrophica

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
100 mM Tris-HCl buffer Hoeflea phototrophica

pH Range

pH Minimum pH Maximum Comment Organism
6 9 more than 70% of the maximum activity is retained between pH 6.0 and 9.0 Hoeflea phototrophica

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
40.48
-
benzoylacetonitrile pH 8.5, 50°C Hoeflea phototrophica