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Literature summary extracted from

  • Miao, Y.; Metzner, R.; Asano, Y.
    Kemp elimination catalyzed by naturally occurring aldoxime dehydratases (2017), ChemBioChem, 18, 451-454 .
    View publication on PubMed

Application

EC Number Application Comment Organism
4.8.1.2 synthesis enzyme is able to catalyze the non-natural Kemp elimination reaction Pseudomonas chlororaphis
4.8.1.2 synthesis enzyme is able to catalyze the non-natural Kemp elimination reaction Bacillus sp. OxB-1
4.8.1.2 synthesis enzyme is able to catalyze the non-natural Kemp elimination reaction Rhodococcus sp. N-771

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.8.1.2 expression in Escherichia coli Pseudomonas chlororaphis
4.8.1.2 expression in Escherichia coli Bacillus sp. OxB-1
4.8.1.2 expression in Escherichia coli Rhodococcus sp. N-771

Protein Variants

EC Number Protein Variants Comment Organism
4.8.1.2 F289A about 45% of wild-type activity Bacillus sp. OxB-1
4.8.1.2 H282A about 5% of wild-type activity Bacillus sp. OxB-1
4.8.1.2 H306A about 3% of wild-type activity Bacillus sp. OxB-1
4.8.1.2 L128A about 30% of wild-type activity Bacillus sp. OxB-1
4.8.1.2 Q204A activity similar to wild-type Bacillus sp. OxB-1
4.8.1.2 T202A about 20% of wild-type activity in absence, 50% in presence of dithionite Bacillus sp. OxB-1

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.8.1.2 0.15
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Pseudomonas chlororaphis
4.8.1.2 0.26
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Rhodococcus sp. N-771
4.8.1.2 0.55
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Bacillus sp. OxB-1
4.8.1.2 0.56
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Rhodococcus sp. N-771
4.8.1.2 0.81
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Pseudomonas chlororaphis
4.8.1.2 0.84
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Bacillus sp. OxB-1

Organism

EC Number Organism UniProt Comment Textmining
4.8.1.2 Bacillus sp. OxB-1 A0A0A8JIJ3
-
-
4.8.1.2 Pseudomonas chlororaphis Q7WSJ4
-
-
4.8.1.2 Rhodococcus sp. N-771 Q76K71
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.8.1.2 5-nitro-1,2-benzoxazole
-
 Pseudomonas chlororaphis 2-hydroxy-5-nitrobenzonitrile
-
 ?
4.8.1.2 5-nitro-1,2-benzoxazole
-
 Bacillus sp. OxB-1 2-hydroxy-5-nitrobenzonitrile
-
 ?
4.8.1.2 5-nitro-1,2-benzoxazole
-
 Rhodococcus sp. N-771 2-hydroxy-5-nitrobenzonitrile
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.8.1.2 OxdB
-
 Bacillus sp. OxB-1

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.8.1.2 0.08
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Rhodococcus sp. N-771
4.8.1.2 0.14
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Bacillus sp. OxB-1
4.8.1.2 0.18
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Pseudomonas chlororaphis
4.8.1.2 9.58
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Bacillus sp. OxB-1
4.8.1.2 9.97
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Pseudomonas chlororaphis
4.8.1.2 17.03
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Rhodococcus sp. N-771

Cofactor

EC Number Cofactor Comment Organism Structure
4.8.1.2 heme the heme iron after purification is primarily in the ferric form. For Kemp elimination, the enzymatic rate over background with ferrous enzyme is approximately 10fold higher than for ferric enzyme Pseudomonas chlororaphis
4.8.1.2 heme the heme iron after purification is primarily in the ferric form. For Kemp elimination, the enzymatic rate over background with ferrous enzyme is approximately 10fold higher than for ferric enzyme Bacillus sp. OxB-1
4.8.1.2 heme the heme iron after purification is primarily in the ferric form. For Kemp elimination, the enzymatic rate over background with ferrous enzyme is approximately 10fold higher than for ferric enzyme Rhodococcus sp. N-771

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.8.1.2 0.143
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Rhodococcus sp. N-771
4.8.1.2 0.216
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Pseudomonas chlororaphis
4.8.1.2 0.227
-
 5-nitro-1,2-benzoxazole ferric oxidation state, pH 7.0, 23°C Bacillus sp. OxB-1
4.8.1.2 11.46
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Bacillus sp. OxB-1
4.8.1.2 65.57
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Rhodococcus sp. N-771
4.8.1.2 66.34
-
 5-nitro-1,2-benzoxazole ferrous oxidation state, pH 7.0, 23°C Pseudomonas chlororaphis