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Literature summary extracted from

  • Maxel, S.; Zhang, L.; King, E.; Acosta, A.; Luo, R.; Li, H.
    In vivo, high-throughput selection of thermostable cyclohexanone monooxygenase (Chmo) (2020), Catalysts, 10, 935 .
No PubMed abstract available

Application

EC Number Application Comment Organism
1.14.13.22 analysis application of an aerobic, high-throughput growth selection platform in Escherichia coli for the discovery of thermostability enhancing mutations for CHMO. The selection employs growth for the easy readout of CHMO activity in vivo, by requiring nicotinamide adenine dinucleotide phosphate (NADPH)-consuming enzymes to restore cellular redox balance Acinetobacter sp.

Protein Variants

EC Number Protein Variants Comment Organism
1.14.13.22 A245G/A288V variant retains native activity, exhibited about 4.4fold improvement in residual activity after 30°C incubation, and demonstrates about 5fold higher cyclohexanone conversion at 37°C compared to the wild-type Acinetobacter sp.
1.14.13.22 A245G/A288V/T415C variant shows improved thermostability at 45°C Acinetobacter sp.

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.13.22 0.0041
-
 cyclohexanone wild-type, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 0.0056
-
 cyclohexanone mutant A245G/A288V, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 0.0366
-
 NADPH wild-type, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 0.0433
-
 NADPH mutant A245G/A288V, pH 7.7, 25°C Acinetobacter sp.

Organism

EC Number Organism UniProt Comment Textmining
1.14.13.22 Acinetobacter sp. P12015
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.13.22 cyclohexanone + NADPH + H+ + O2
-
 Acinetobacter sp. hexano-6-lactone + NADP+ + H2O
-
 ?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.13.22 9.67
-
 NADPH mutant A245G/A288V, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 11.6
-
 cyclohexanone mutant A245G/A288V, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 12.9
-
 cyclohexanone wild-type, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 13.5
-
 NADPH wild-type, pH 7.7, 25°C Acinetobacter sp.

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.13.22 220
-
 NADPH mutant A245G/A288V, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 370
-
 NADPH wild-type, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 2050
-
 cyclohexanone mutant A245G/A288V, pH 7.7, 25°C Acinetobacter sp.
1.14.13.22 3150
-
 cyclohexanone wild-type, pH 7.7, 25°C Acinetobacter sp.