Cyclohexanone monooxygenase from Acinetobacter johnsonii

Enzyme Description

Species
Acinetobacter johnsonii β†—
Extremophile
No
EC Number
1.14.13.22 β†—

Sequence

Length: 543 amino acids
MSQKMDFDAIVIGGGFGGLYAVKKLRDELELKVQAFDKATDVAGTWYWNRYPGALTDTETHLYCYSWDKELLQSLEIKKKYVQGPDVRKYLQQVAEKHDLKKSYQFNTAVQSAHYNEADALWEVTTEYGDKYTARFLITALGLLSAPNLPNIKGINQFKGELHHTSRWPDDVSFEGKRVGVIGTGSTGVQVITAVAPLAKHLTVFQRSAQYSVPIGNDPLSEEDVKKIKDNYDKSLGWCMNSALAFALNESTVPAMSVSAEERKAVFEKAWQTGGGFRFMFETFGDIATNMEANIEAQNFIKGKIAEIVKDPAIAQKLMPQDLYAKRPLCDSGYYNTFNRDNVRLEDVKANPIVEITENGVKLENGDFVELDMLICATGFDAVDGNYVRMDIQGKNGLAMKDYWKEGPSSYMGVTVNNYPNMFMVLGPNGPFTNLPPSIESQVEWISDTIQYTVENNVESIEATKEAEEQWTQTCANIAEMTLFPKAQSWIFGANIPGKKNTVYFYLGGLKEYRTCASNCKNHAYEGFDIQLQRSDIKQPANA
Severin Wedde et al. (2017) β€” High-Throughput Feasible Screening Tool for Determining Enzyme Stabilities against Organic Solvents Directly from Crude Extracts
ChemBioChem  Β· doi:10.1002/cbic.201700526 β†—  Β· Stability - Incubation Stability - Tm
8 measurements ▼
Database ID
UniProt: P12015 β†—
Sequence Annotation
Explicit - Provided
Protein Source
Recombinant, host bacterium Escherichia coli BL21 (DE3)

Experimental Data (8 measurements)

8 measurements
Property Assay Solvent Solvent Volume Aqueous Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
Stability - Incubation Activity after incubation (4 hours at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Cyclohexane 10% 85.7 83.2 % Incubation: 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚, Assay: 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚ Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature Cyclohexanone, Molecular oxygen H2O , Ξ΅-caprolactone 0.375 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Unclear temperature, assay in diluted organic solvent here classified as aqueous phase
Stability - Incubation Activity after incubation (4 hours at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Isooctane 10% 85.7 83.0 % Incubation: 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚, Assay: 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚ Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature Cyclohexanone, Molecular oxygen H2O , Ξ΅-caprolactone 0.375 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Unclear temperature, assay in diluted organic solvent here classified as aqueous phase
Stability - Incubation Activity after incubation (4 hours at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase Methylcyclohexane 10% 85.7 84.8 % Incubation: 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚, Assay: 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚ Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature Cyclohexanone, Molecular oxygen H2O , Ξ΅-caprolactone 0.375 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Unclear temperature, assay in diluted organic solvent here classified as aqueous phase
Stability - Incubation Activity after incubation (4 hours at room temperature), measured by absorbance spectrophotometry (NADPH absorbance measurement, 340 nm) in aqueous phase tert-Butyl Methyl Ether (MTBE) 10% 85.7 80.9 % Incubation: 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚, Assay: 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚ Incubation: 7, Assay: 7 Incubation: Room temperature, Assay: Room temperature Cyclohexanone, Molecular oxygen H2O , Ξ΅-caprolactone 0.375 mM NADPH β€” Water-incubated control (in %), assay in aqueous phase | Unclear temperature, assay in diluted organic solvent here classified as aqueous phase
Stability - Tm Tm measured by NanoDSF in the presence of organic solvent Cyclohexane 10% 39.3 38.7 Β°C 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚ 7 β€” β€” β€” β€” β€” Classical aqueous control (in Β°C)
Stability - Tm Tm measured by NanoDSF in the presence of organic solvent Isooctane 10% 39.3 39.0 Β°C 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚ 7 β€” β€” β€” β€” β€” Classical aqueous control (in Β°C)
Stability - Tm Tm measured by NanoDSF in the presence of organic solvent Methylcyclohexane 10% 39.3 39.0 Β°C 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚ 7 β€” β€” β€” β€” β€” Classical aqueous control (in Β°C)
Stability - Tm Tm measured by NanoDSF in the presence of organic solvent tert-Butyl Methyl Ether (MTBE) 10% 39.3 35.2 Β°C 50 mM potassium phosphate buffer, 200 mg/L MgClβ‚‚ 7 β€” β€” β€” β€” β€” Classical aqueous control (in Β°C)

Visualization : Stability β€” Tm

One bar per measurement. Colour = solvent, shade = solvent volume.

Visualization : Stability β€” Incubation

One bar per measurement. Colour = solvent, shade = solvent volume.

Structure

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