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
233 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 (233 measurements)

233 measurements β€” page 8 of 12
Mutation Property Assay Solvent Solvent Volume Aqueous ReferenceWT Reference Measured Value Units Extraction method Data type 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 % v/v 100 9 31 % Digital Continuous ? ? 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 % v/v 100 4 0 % Digital Continuous ? ? 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 % v/v 100 83 95 % Digital Continuous ? ? 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 % v/v 100 85 78 % Digital Continuous ? ? 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 % v/v 100 68 45 % Digital Continuous ? ? 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 % v/v 100 42 18 % Digital Continuous ? ? 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 % v/v 100 23 5 % Digital Continuous ? ? 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 % v/v 100 9 3 % Digital Continuous ? ? 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 % v/v 100 83 90 % Digital Continuous ? ? 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 % v/v 100 85 95 % Digital Continuous ? ? 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 % v/v 100 68 64 % Digital Continuous ? ? 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 % v/v 100 42 48 % Digital Continuous ? ? 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 % v/v 100 23 28 % Digital Continuous ? ? 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 % v/v 100 9 16 % Digital Continuous ? ? 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 % v/v 100 4 0 % Digital Continuous ? ? 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 % v/v 100 98 101 % Digital Continuous ? ? 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 % v/v 100 97 107 % Digital Continuous ? ? 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 % v/v 100 94 110 % Digital Continuous ? ? 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 % v/v 100 91 109 % Digital Continuous ? ? 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 % v/v 100 76 111 % Digital Continuous ? ? 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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