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 4 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
F87A/T235A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 5% 100 113 86 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100 110 64 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 15% 100 106 47 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100 78 29 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 25% 100 43 9 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100 16 4 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 5% 100 113 98 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100 110 72 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 15% 100 106 50 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100 78 29 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 25% 100 43 16 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024R Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100 16 0 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024T Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 5% 100 113 90 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024T Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 10% 100 110 76 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024T Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 15% 100 106 54 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024T Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 20% 100 78 28 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024T Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 25% 100 43 15 % ? ? Room temperature 0.5 mM p-Nitrophenoxylauric Acid Hydroxylated fatty acid , p-Nitrophenol ? mM NADP+ β€” Classical aqueous control (in %)
F87A/T235A/R471A/S1024T Activity - Classical Activity measured by absorbance spectrophotometry (p-nitrophenol absorbance measurement, 410 nm) in the presence of organic solvent Dimethyl Sulfoxide (DMSO) 30% 100 16 0 % ? ? 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 Dimethyl Sulfoxide (DMSO) 5% 100 113 99 % ? ? 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 Dimethyl Sulfoxide (DMSO) 10% 100 110 80 % ? ? 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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