Characterization of a catalytically self-sufficient 119,000-dalton cytochrome P-450 monooxygenase induced by barbiturates in Bacillus megaterium.

A unique cytochrome P-450-dependent fatty acid monooxygenase from Bacillus megaterium ATCC 14581 is strongly induced by phenobarbital (Narhi, L. O., and Fulco, A. J. (1982) J. Biol. Chem. 257, 2147-2150) and many other barbiturates (Kim, B.-H., and Fulco, A. J. (1983) Biochem. Biophys. Res. Commun. 116, 843-850). This monooxygenase has now been purified to homogeneity from pentobarbital-induced bacteria as a single polypeptide with a molecular weight of 119,000 +/- 5,000 daltons. In the presence of NADPH and O2, it can catalyze the oxygenation of long chain fatty acids without the aid of any other protein. The enzyme has a catalytic center activity of 4,600 nmol of fatty acid oxygenated per nmol of P-450 (the highest activity yet reported for a P-450-dependent monooxygenase) and also functions as a highly active cytochrome c reductase in the presence of NADPH. The purified holoenzyme is a soluble protein containing 40 mol % hydrophobic amino acid residues and 1 mol each of FAD and FMN/mol of heme. It is isolated and purified in the low spin form but is converted to the high spin form in the presence of long chain fatty acids. The enzyme, which catalyzes the omega-2 hydroxylation of saturated fatty acids and the hydroxylation and epoxidation of unsaturated fatty acids has its highest affinity (Km = 2 +/- 1 microM) for the C15 and C16 chain lengths.

• Publication year

  1986

• Venue

  Journal of Biological Chemistry

• Publication date

  1986-06-05

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0021-9258(17)38369-2PMID 3086309
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Purification and characterization of pentobarbital-induced cytochrome P-450BM-1 from Bacillus megaterium ATCC 14581.

  1985cited by this paper
• Induction of a cytochrome P-450-dependent fatty acid monooxygenase in Bacillus megaterium by a barbiturate analog, 1-[2-phenylbutyryl]-3-methylurea

  1985cited by this paper
• Acylureas: a new class of barbiturate-like bacterial cytochrome P-450 inducers.

  1984cited by this paper
• Induction by barbiturates of a cytochrome P-450-dependent fatty acid monooxygenase in Bacillus megaterium: relationship between barbiturate structure and inducer activity.

  1983cited by this paper
• Proteolytic cleavage of Hansenula anomala flavocytochrome b2 into its two functional domains. Isolation of a highly active flavodehydrogenase and a cytochrome b2 core.

  1983cited by this paper
• Partial characterization of a barbiturate-induced cytochrome P-450-dependent fatty acid monooxygenase from Bacillus megaterium.

  1983cited by this paper
• Phenobarbital induction of a soluble cytochrome P-450-dependent fatty acid monooxygenase in Bacillus megaterium.

  1982cited by this paper
• Steroidogenic electron transport in adrenal cortex mitochondria

  1982cited by this paper
• Studies on the substrate specificity and inducibility of cytochrome P-450meg.

  1981cited by this paper
• Epoxidation of unsaturated fatty acids by a soluble cytochrome P-450-dependent system from Bacillus megaterium.

  1981cited by this paper
• Genetic mechanisms controlling the induction of polysubstrate monooxygenase (P-450) activities.

  1981cited by this paper
• Separate roles for FMN and FAD in catalysis by liver microsomal NADPH-cytochrome P-450 reductase.

  1981cited by this paper
• Studies on the microsomal mixed-function oxidase system: mechanism of action of hepatic NADPH-cytochrome P-450 reductase.

  1981cited by this paper
• A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels.

  1980cited by this paper
• Oxygen activation by cytochrome P-450.

  1980cited by this paper
• Purification and characterization of cytochrome P-450meg.

  1979cited by this paper
• Purified liver microsomal NADPH-cytochrome P-450 reductase. Spectral characterization of oxidation-reduction states.

  1978cited by this paper
• The Tools of Biochemistry

  1977cited by this paper
• Characteristics of a cytochrome P-450-dependent fatty acid omega-2 hydroxylase from bacillus megaterium.

  1977cited by this paper
• Involvement of a single hydroxylase species in the hydroxylation of palmitate at the omega-1, omega-2 and omega-3 positions by a preparation from Bacillus megaterium.

  1976cited by this paper
• Characterization of a cytochrome P-450-dependent steroid hydroxylase system present in Bacillus megaterium.

  1976cited by this paper
• Omega-1, Omega-2 and Omega-3 hydroxylation of long-chain fatty acids, amides and alcohols by a soluble enzyme system from Bacillus megaterium.

  1975cited by this paper
• Preparation of homogeneous NADPH-cytochrome P-450 reductase from rat liver.

  1975cited by this paper
• A rapid micromethod for determination of FMN and FAD in mixtures.

  1973cited by this paper
• Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

  1970cited by this paper
• THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE.

  1964cited by this paper

• Multiple roles for iron in microbial physiology: Bacterial oxygen sensing by heme-based sensors.

  2025cites this paper
• Unique electron transfer system of cytochrome P450 monooxygenase includes a mechanism of fatty acid β-oxidation

  2025cites this paper
• Protein Engineering for Enhancing Electron Transfer in P450‐Mediated Catalysis

  2025influential citation
• AFM-Based Monitoring of Enzymatic Activity of Individual Molecules of Cytochrome CYP102A1

  2025cites this paper
• Hydroxylation and Biological Activity Evaluation of Isosteviol C12 by P450 BM3

  2025cites this paper
• Full-length structure of CPR-containing self-sufficient cytochrome P450

  2025cites this paper
• Recyclable Whole-Cell Biotransformation System for the Direct Hydroxylation of Propane Catalyzed by a Robust Intracellular Wild-Type Cytochrome P450BM3 Activated by Decoy Molecules

  2025cites this paper
• Genetically engineered bacteria and microalgae expressing a mutant of cytochrome P450 BM3 for efficient Diuron degradation in wastewater treatment

  2025cites this paper
• Structure–Function Analysis of the Self-Sufficient CYP102 Family Provides New Insights into Their Biochemistry

  2025cites this paper
• Strategies found not to be suitable for stabilizing high steroid hydroxylation activities of CYP450 BM3-based whole-cell biocatalysts

  2024cites this paper
• Choose Your Own Adventure: A Comprehensive Database of Reactions Catalyzed by Cytochrome P450 BM3 Variants

  2024cites this paper
• Tuning architectural organization of eukaryotic P450 system to boost bioproduction in Escherichia coli

  2024cites this paper
• Protein fusion strategies for a multi-component Rieske oxygenase

  2024cites this paper
• Pharmaceutical removal from wastewater by introducing cytochrome P450s into microalgae

  2024cites this paper
• Multidomain chimeric enzymes as a promising alternative for biocatalysts improvement: a minireview

  2024cites this paper
• Solid-State Nanopore-Based Nanosystem for Registration of Enzymatic Activity of a Single Molecule of Cytochrome P450 BM3

  2024cites this paper
• Harnessing heme chemistry: Recent advances in the biocatalytic applications of cytochrome P450 monooxgenases

  2023cites this paper
• Tryptophan extends the life of cytochrome P450

  2023cites this paper
• The Versatile Biocatalyst of Cytochrome P450 CYP102A1: Structure, Function, and Engineering

  2023cites this paper
• CYP108N14: A Monoterpene Monooxygenase from Rhodococcus globerulus.

  2023cites this paper
• Catalytic and Electrocatalytic Mechanisms of Cytochromes P450 in the Development of Biosensors and Bioreactors

  2023cites this paper
• Green Oxidative Catalytic Processes for the Preparation of APIs and Precursors

  2023cites this paper
• Cytochrome P450 for environmental remediation: catalytic mechanism, engineering strategies and future prospects

  2023cites this paper
• Bioengineering for the Microbial Degradation of Petroleum Hydrocarbon Contaminants

  2023cites this paper
• Direct aromatic nitration by bacterial P450 enzymes.

  2023cites this paper
• Hydrophobic Outer Membrane Pores Boost Testosterone Hydroxylation by Cytochrome P450 BM3 Containing Cells

  2022cites this paper
• Oleic acid based experimental evolution of Bacillus megaterium yielding an enhanced P450 BM3 variant

  2022cites this paper
• Opportunities for Accelerating Drug Discovery and Development by Using Engineered Drug-Metabolizing Enzymes

  2022cites this paper
• Molecular Basis for a Toluene Monooxygenase to Govern Substrate Selectivity

  2022cites this paper
• Unravelling the role of transient redox partner complexes in P450 electron transfer mechanics

  2022cites this paper
• Engineering Catalytically Self-Sufficient P450s.

  2022cites this paper
• Carving the Active Site of CYP153A7 Monooxygenase for Improving Terminal Hydroxylation of Medium‐Chain Fatty Acids

  2022cites this paper
• Cytochromes P450 in biosensing and biosynthesis applications: Recent progress and future perspectives

  2022cites this paper
• Approaches for increasing the electrocatalitic efficiency of cytochrome P450 3A4.

  2022cites this paper
• Roles of Ferredoxin-NADP+ Oxidoreductase and Flavodoxin in NAD(P)H-Dependent Electron Transfer Systems

  2022influential citation
• Expanding the Substrate Scope of Nitrating Cytochrome P450 TxtE by Active Site Engineering of a Reductase Fusion

  2021cites this paper
• AFM and FTIR Investigation of the Effect of Water Flow on Horseradish Peroxidase

  2021cites this paper
• Biocatalytic One-Carbon Ring Expansion of Aziridines to Azetidines via a Highly Enantioselective [1,2]-Stevens Rearrangement.

  2021cites this paper
• Identification and characterization of a novel class of self-sufficient cytochrome P450 hydroxylase involved in cyclohexanecarboxylate degradation in Paraburkholderia terrae strain KU-64.

  2021cites this paper
• Enzyme modification using mutation site prediction method for enhancing the regioselectivity of substrate reaction sites

  2021cites this paper
• Promising optimization of bacterial cytochrome P450BM3 enzyme production by engineered Escherichia coli BL21

  2021cites this paper
• The “beauty in the beast”—the multiple uses of Priestia megaterium in biotechnology

  2021influential citation
• Engineering the biomimetic cofactors of NMNH for cytochrome P450 BM3 based on binding conformation refinement

  2021cites this paper
• Decision letter for "Optimization of operation conditions for improved cytochrome P450BM3 enzymatic reaction yield"

  2021cites this paper
• A Promiscuous Bacterial P450: The Unparalleled Diversity of BM3 in Pharmaceutical Metabolism

  2021cites this paper
• De novo biosynthesis of bioactive isoflavonoids by engineered yeast cell factories

  2021cites this paper
• Enantioselective Bioamination of Aromatic Alkanes Using Ammonia: A Multienzymatic Cascade Approach

  2020cites this paper
• Bacterial cytochrome P450-catalyzed regio- and stereoselective steroid hydroxylation enabled by directed evolution and rational design

  2020cites this paper
• Methylene Oxidation of Alkyl Sulfates by Cytochrome P450BM-3 and a Role for Conformational Selection in Substrate Recognition.

  2020cites this paper
• Advanced Understanding of the Electron Transfer Pathway of Cytochrome P450s

  2020cites this paper
• Structure-based mechanisms: On the way to apply alcohol dehydrogenases/reductases to organic-aqueous systems.

  2020cites this paper
• Systematic Evolution of Decoy Molecules for the Highly Efficient Hydroxylation of Benzene and Small Alkanes Catalyzed by Wild-Type Cytochrome P450BM3

  2020cites this paper
• Cytochromes P450 (P450s): A review of the class system with a focus on prokaryotic P450s.

  2020cites this paper
• Regioselective Hydroxylation of Naringin Dihydrochalcone to Produce Neoeriocitrin Dihydrochalcone by CYP102A1 (BM3) Mutants

  2020cites this paper
• Surface charge-controlled electron transfer and catalytic behavior of immobilized cytochrome P450 BM3 inside dendritic mesoporous silica nanoparticles

  2020cites this paper
• A BIOPHYSICAL INVESTIGATION OF STABILITY, LIGAND BINDING, AND IRON STATE OF CYP102A1

  2020cites this paper
• Directed Evolution of an FeII-Dependent Halogenase for Asymmetric C(sp3)–H Chlorination

  2019cites this paper
• Statistical Analysis of the Benefits of Focused Saturation Mutagenesis in Directed Evolution Based on Reduced Amino Acid Alphabets

  2019cites this paper
• Production of metabolites of the anti-cancer drug noscapine using a P450BM3 mutant library

  2019cites this paper
• CYP52A23 from Candida albicans and its Substrate Preference for Fatty Acid Hydroxylation.

  2019cites this paper
• Gut Reactions: Breaking Down Xenobiotic–Microbiome Interactions

  2019cites this paper
• P450BM3-Catalyzed Oxidations Employing Dual Functional Small Molecules

  2019cites this paper
• Multi-Functional Oxidase Activity of CYP102A1 (P450BM3) in the Oxidation of Quinolines and Tetrahydroquinolines.

  2019cites this paper
• Broadening the Scope of Biocatalysis in Sustainable Organic Synthesis.

  2019cites this paper
• The Registration of a Biomaser-Like Effect in an Enzyme System with an RTM Sensor

  2019cites this paper
• Cytochrome P450 research and The Journal of Biological Chemistry

  2019cites this paper
• Cytochrome P450 Monooxygenases in Biotechnology and Synthetic Biology.

  2019cites this paper
• Regioselective hydroxylation pathway of tenatoprazole to produce human metabolites by Bacillus megaterium CYP102A1

  2019cites this paper
• An integrated screening system for the selection of exemplary substrates for natural and engineered cytochrome P450s

  2019cites this paper
• The Recent Developments of Enzymatic Oxidation

  2019cites this paper
• Exploring the Biocatalytic Potential of a Self‐Sufficient Cytochrome P450 from Thermothelomyces thermophila

  2019cites this paper
• In vitro characterization of CYP102G4 from Streptomyces cattleya: A self-sufficient P450 naturally producing indigo.

  2018cites this paper
• One-Pot Biocombinatorial Synthesis of Herbicidal Thaxtomins

  2018cites this paper
• Co-immobilization of P450 BM3 and glucose dehydrogenase on different supports for application as a self-sufficient oxidative biocatalyst

  2018cites this paper
• Biochemical Characterization of CYP505D6, a Self-Sufficient Cytochrome P450 from the White-Rot Fungus Phanerochaete chrysosporium

  2018cites this paper
• Harnessing P450 enzymes as biocatalysts for selective C-H bond hydroxylation

  2018cites this paper
• Characterization of Cytochrome P450 Enzymes and Their Applications in Synthetic Biology.

  2018cites this paper
• Enzyme Fusions in Biocatalysis: Coupling Reactions by Pairing Enzymes

  2018cites this paper
• Chemoenzymatic Route to Oxyfunctionalized Cembranoids Facilitated by Substrate and Protein Engineering.

  2018cites this paper
• Atomic Force Microscopy for Protein Detection and Their Physicoсhemical Characterization

  2018cites this paper
• Engineering a self-sufficient Mycobacterium tuberculosis CYP130 by gene fusion with the reductase-domain of CYP102A1 from Bacillus megaterium.

  2018cites this paper
• The full-length cytochrome P450 enzyme CYP102A1 dimerizes at its reductase domains and has flexible heme domains for efficient catalysis

  2018cites this paper
• Enzymatic construction of highly strained carbocycles

  2018cites this paper
• General aspects of cytochrome P 450 monooxygenases Cytochrome

  2018cites this paper
• Exploring PTDH–P450BM3 Variants for the Synthesis of Drug Metabolites

  2018cites this paper
• Cloning, expression and characterisation of P450-Hal1 (CYP116B62) from Halomonas sp. NCIMB 172: A self-sufficient P450 with high expression and diverse substrate scope.

  2018cites this paper
• Stabilization of the Reductase Domain in the Catalytically Self‐Sufficient Cytochrome P450BM3 by Consensus‐Guided Mutagenesis

  2018cites this paper
• Effects of Alternative Redox Partners and Oxidizing Agents on CYP154C8 Catalytic Activity and Product Distribution

  2018cites this paper
• Enzymatic synthesis and chemical inversion provide both enantiomers of bioactive epoxydocosapentaenoic acids[S]

  2018cites this paper
• Characterisation of CYP102A25 from Bacillus marmarensis and CYP102A26 from Pontibacillus halophilus: P450 Homologues of BM3 with Preference towards Hydroxylation of Medium‐Chain Fatty Acids

  2018cites this paper
• Ultrasensitive nanowire-based detection of HCVcoreAg in the serum using a microwave generator

  2018cites this paper
• Metabolic Engineering for Enhanced Medium Chain Omega Hydroxy Fatty Acid Production in Escherichia coli

  2018cites this paper
• Crystallographic insights into a cobalt (III) sepulchrate based alternative cofactor system of P450 BM3 monooxygenase.

  2018cites this paper
• Construction and characterization of engineered diesel-degrading bacterium

  2017influential citation
• Influence of lipid membrane environment on the kinetics of the cytochrome P450 reductase- cytochrome P450 3A4 enzyme system in nanodiscs

  2017cites this paper
• Novel 96-well quantitative bioelectrocatalytic analysis platform reveals highly efficient direct electrode regeneration of cytochrome P450 BM3 on indium tin oxide.

  2017influential citation
• Direct Hydroxylation of Benzene to Phenol by Cytochrome P450BM3 Triggered by Amino Acid Derivatives.

  2017cites this paper
• Fragment Profiling Approach to Inhibitors of the Orphan M. tuberculosis P450 CYP144A1.

  2017cites this paper
• Strategies for the construction of insect P450 fusion enzymes

  2017cites this paper
• Expression, Purification, and Biochemical Characterization of the Flavocytochrome P450 CYP505A30 from Myceliophthora thermophila

  2017cites this paper