Cytochrome P450 BM-3 from Bacillus megaterium

Enzyme Description

Species
Bacillus megaterium β†—
Extremophile
No
EC Number
1.14.14.1 β†—

Sequence

Length: 1048 amino acids
TIKEMPQPKTFGELKNLPLLNTDKPVQALMKIADELGEIFKFEAPGRVTRYLSSQRLIKEACDESRFDKNLSQALKFVRDFAGDGLFTSWTHEKNWKKAHNILLPSFSQQAMKGYHAMMVDIAVQLVQKWERLNADEHIEVPEDMTRLTLDTIGLCGFNYRFNSFYRDQPHPFITSMVRALDEAMNKLQRANPDDPAYDENKRQFQEDIKVMNDLVDKIIADRKASGEQSDDLLTHMLNGKDPETGEPLDDENIRYQIITFLIAGHETTSGLLSFALYFLVKNPHVLQKAAEEAARVLVDPVPSYKQVKQLKYVGMVLNEALRLWPTAPAFSLYAKEDTVLGGEYPLEKGDELMVLIPQLHRDKTIWGDDVEEFRPERFENPSAIPQHAFKPFGNGQRACIGQQFALHEATLVLGMMLKHFDFEDHTNYELDIKETLTLKPEGFVVKAKSKKIPLGGIPSPSTEQSAKKVRKKAENAHNTPLLVLYGSNMGTAEGTARDLADIAMSKGFAPQVATLDSHAGNLPREGAVLIVTASYNGHPPDNAKQFVDWLDQASADEVKGVRYSVFGCGDKNWATTYQKVPAFIDETLAAKGAENIADRGEADASDDFEGTYEEWREHMWSDVAAYFNLDIENSEDNKSTLSLQFVDSAADMPLAKMHGAFSTNVVASKELQQPGSARSTRHLEIELPKEASYQEGDHLGVIPRNYEGIVNRVTARFGLDASQQIRLEAEEEKLAHLPLAKTVSVEELLQYVELQDPVTRTQLRAMAAKTVCPPHKVELEALLEKQAYKEQVLAKRLTMLELLEKYPACEMKFSEFIALLPSIRPRYYSISSSPRVDEKQASITVSVVSGEAWSGYGEYKGIASNYLAELQEGDTITCFISTPQSEFTLPKDPETPLIMVGPGTGVAPFRGFVQARKQLKEQGQSLGEAHLYFGCRSPHEDYLYQEELENAQSEGIITLHTAFSRMPNQPKTYVQHVMEQDGKKLIELLDQGAHFYICGDGSQMAPAVEATLMKSYADVHQVSEADARLWLQQLEEKGRYAKDVWAG
Tuck Seng Wong et al. (2003) β€” Laboratory evolution of cytochrome P450 BM-3 monooxygenase for organic cosolvents
Biotechnology and Bioengineering  Β· doi:10.1002/bit.10896 β†—  Β· Activity - Classical
253 measurements ▼
Database ID
UniProt: P14779 β†—
Sequence Annotation
Inferred - from protein name (first residue from UniProt sequence absent from the mature protein)
Protein Source
Recombinant, host bacterium Escherichia coli

Experimental Data (253 measurements)

253 measurements β€” page 9 of 13
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Solution pH Temperature Substrate(s) Product(s) Cofactor(s) Shaking Comments
T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 12.5% 100 9 31 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 15% 100 4 0 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 2% 100 83 95 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 3% 100 85 78 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 5% 100 68 45 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 7.5% 100 42 18 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 10% 100 23 5 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 12.5% 100 9 3 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 2% 100 83 90 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 3% 100 85 95 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 5% 100 68 64 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 7.5% 100 42 48 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 10% 100 23 28 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 12.5% 100 9 16 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Acetone 15% 100 4 0 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethylformamide (DMF) 1% 100 98 101 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethylformamide (DMF) 1.5% 100 97 107 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethylformamide (DMF) 2% 100 94 110 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethylformamide (DMF) 3% 100 91 109 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
T235A/R471A/E494K/S1024E Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethylformamide (DMF) 4% 100 76 111 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
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Mutations in this dataset (16)

WT F87A F87A/T235A/R471A/E494K/S1024E F87A/T235A/R471A/S1024R F87A/T235A/R471A/S1024T F87A/T235A/S1024R T235A/R471A/E494K/S1024 T235A/R471A/E494K/S1024E T235A/R471A/E494K/S1025 T235A/R471A/E494K/S1026 T235A/R471A/E494K/S1027 T235A/R471A/E494K/S1028 T235A/R471A/E494K/S1029 T235A/R471A/S1024R T235A/R471A/S1024T T235A/S1024R

Visualization : Activity β€” Classical

One bar per measurement. Colour = solvent, shade = solvent volume. β€” β€” β€” Reference value. Hover for details.

Mutation Effect

Mutation impact on enzyme stability and function in the presence of organic solvent: comparison of wild-type and mutant values in identical conditions.

Structure

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