scholarly journals Structural and functional characterisation of the cytochrome P450 enzyme CYP268A2 from Mycobacterium marinum

2018 ◽  
Vol 475 (4) ◽  
pp. 705-722 ◽  
Author(s):  
Stella A. Child ◽  
Elise F. Naumann ◽  
John B. Bruning ◽  
Stephen G. Bell
Keyword(s):  
Fatty Acids ◽  
Cytochrome P450 ◽  
Active Site ◽  
Geranyl Acetate ◽  
Cytochrome P450 Enzyme ◽  
P450 Monooxygenase ◽  
Monooxygenase Activity ◽  
Functional Characterisation ◽  
P450 Enzyme ◽  
Chain Fatty Acids

Members of the cytochrome P450 monooxygenase family CYP268 are found across a broad range of Mycobacterium species including the pathogens Mycobacterium avium, M. colombiense, M. kansasii, and M. marinum. CYP268A2, from M. marinum, which is the first member of this family to be studied, was purified and characterised. CYP268A2 was found to bind a variety of substrates with high affinity, including branched and straight chain fatty acids (C10–C12), acetate esters, and aromatic compounds. The enzyme was also found to bind phenylimidazole inhibitors but not larger azoles, such as ketoconazole. The monooxygenase activity of CYP268A2 was efficiently reconstituted using heterologous electron transfer partner proteins. CYP268A2 hydroxylated geranyl acetate and trans-pseudoionone at a terminal methyl group to yield (2E,6E)-8-hydroxy-3,7-dimethylocta-2,6-dien-1-yl acetate and (3E,5E,9E)-11-hydroxy-6,10-dimethylundeca-3,5,9-trien-2-one, respectively. The X-ray crystal structure of CYP268A2 was solved to a resolution of 2.0 Å with trans-pseudoionone bound in the active site. The overall structure was similar to that of the related phytanic acid monooxygenase CYP124A1 enzyme from Mycobacterium tuberculosis, which shares 41% sequence identity. The active site is predominantly hydrophobic, but includes the Ser99 and Gln209 residues which form hydrogen bonds with the terminal carbonyl group of the pseudoionone. The structure provided an explanation on why CYP268A2 shows a preference for shorter substrates over the longer chain fatty acids which bind to CYP124A1 and the selective nature of the catalysed monooxygenase activity.

scholarly journals Differentiating Keratinocytes Express a Novel Cytochrome P450 Enzyme, CYP2B19, Having Arachidonate Monooxygenase Activity

1998 ◽  
Vol 273 (48) ◽  
pp. 32071-32079 ◽  
Author(s):  
Diane S. Keeney ◽  
Colin Skinner ◽  
Jeffrey B. Travers ◽  
Jorge H. Capdevila ◽  
Lillian B. Nanney ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Cytochrome P450 Enzyme ◽  
Monooxygenase Activity ◽  
P450 Enzyme

scholarly journals Transgenic Production of Epoxy Fatty Acids by Expression of a Cytochrome P450 Enzyme from Euphorbia lagascaeSeed

2002 ◽  
Vol 128 (2) ◽  
pp. 615-624 ◽  
Author(s):  
Edgar B. Cahoon ◽  
Kevin G. Ripp ◽  
Sarah E. Hall ◽  
Brian McGonigle
Keyword(s):  
Fatty Acids ◽  
Cytochrome P450 ◽  
Cytochrome P450 Enzyme ◽  
Epoxy Fatty Acids ◽  
P450 Enzyme

Reversible Pressure Deformation of A Thermophilic Cytochrome P450 Enzyme (CYP119) and Its Active-Site Mutants

2001 ◽  
Vol 123 (15) ◽  
pp. 3412-3417 ◽  
Author(s):  
Richard A. Tschirret-Guth ◽  
Laura S. Koo ◽  
Gaston Hui Bon Hoa ◽  
Paul R. Ortiz de Montellano
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Cytochrome P450 Enzyme ◽  
P450 Enzyme ◽  
Active Site Mutants

scholarly journals The efficient and selective catalytic oxidation of para-substituted cinnamic acid derivatives by the cytochrome P450 monooxygenase, CYP199A4

RSC Advances ◽  
2016 ◽  
Vol 6 (60) ◽  
pp. 55286-55297 ◽  
Author(s):  
Rebecca R. Chao ◽  
James J. De Voss ◽  
Stephen G. Bell
Keyword(s):  
Cytochrome P450 ◽  
Catalytic Oxidation ◽  
Cinnamic Acid ◽  
Product Formation ◽  
Cytochrome P450 Enzyme ◽  
Cinnamic Acids ◽  
P450 Monooxygenase ◽  
Cinnamic Acid Derivatives ◽  
Selective Catalytic Oxidation ◽  
P450 Enzyme

The cytochrome P450 enzyme, CYP199A4 oxidised para substituted alkyloxy- and alkyl-cinnamic acids, with high product formation activity.

scholarly journals A Bifunctional Cytochrome P450 Enzyme Involved in the O-Dealkylation or N-Dealkoxymethylation of Chloroacetanilide Herbicides in Rhodococcus sp. B2

2021 ◽  
Author(s):  
hongming liu ◽  
Meng Yuan ◽  
Aimin Liu ◽  
Lei Ren ◽  
Guo-ping Zhu ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Gene Cluster ◽  
Draft Genome ◽  
Maximal Activity ◽  
Gas Chromatography Mass Spectrometry ◽  
Genome Comparison ◽  
Cytochrome P450 Enzyme ◽  
P450 Monooxygenase ◽  
Emerging Pollutant ◽  
P450 Enzyme

Abstract Background: The chloroacetamide herbicides pretilachlor is an emerging pollutant. Due to the large amount of use, its presence in the environment threatens human health. However, the molecular mechanism of pretilachlor degradation remains unknown. Results: Now, Rhodococcus sp. B2 was isolated from rice field and shown to degrade pretilachlor. The maximum pretilachlor degradation efficiency (86.1%) was observed at a culture time of 5 d, an initial substrate concentration 50 mg/L, pH 6.98, and 30.1°C. One novel metabolite N-hydroxyethyl-2-chloro-N-(2, 6-diethyl-phenyl)-acetamide was identified by gas chromatography-mass spectrometry (GC-MS). Draft genome comparison demonstrated that a 32,147-bp DNA fragment, harboring gene cluster (EthRABCDB2), was absent from the mutant strain TB2 which could not degrade pretilachlor. The Eth gene cluster, encodes an AraC/XylS family transcriptional regulator (EthRB2), a ferredoxin reductase (EthAB2), a cytochrome P450 monooxygenase (EthBB2), a ferredoxin (EthCB2) and a 10-kDa protein of unknown function (EthDB2). Complementation with EthABCDB2 and EthABDB2, but not EthABCB2 in strain TB2 restored its ability to degrade chloroacetamide herbicides. Subsequently, codon optimization of EthABCDB2 was performed, after which the optimized components were separately expressed in Escherichia coli, and purified using Ni-affinity chromatography. A mixture of EthABCDB2 or EthABDB2 but not EthABCB2 catalyzed the N-dealkoxymethylation of alachlor, acetochlor, butachlor, and propisochlor and O-dealkylation of pretilachlor, revealing that EthD acted as a ferredoxin in strain B2. EthABDB2 displayed maximal activity at 30 °C and pH 7.5. Conclusions: This is the first report of a P450 family oxygenase catalyzing the O-dealkylation and N-dealkoxymethylation of pretilachlor and propisochlor, respectively. And the results of the present study provide a microbial resource for the remediation of chloroacetamide herbicides-contaminated sites.

scholarly journals A Bifunctional Cytochrome P450 Enzyme Involved in the O-Dealkylation and N-Dealkoxymethylation toward Chloroacetanilide Herbicides in Rhodococcus sp. B2

2021 ◽  
Author(s):  
hongming liu ◽  
Meng Yuan ◽  
Aimin Liu ◽  
Guo-ping Zhu ◽  
Li-na Sun
Keyword(s):  
Cytochrome P450 ◽  
Gene Cluster ◽  
Draft Genome ◽  
Maximal Activity ◽  
Gas Chromatography Mass Spectrometry ◽  
Genome Comparison ◽  
Cytochrome P450 Enzyme ◽  
P450 Monooxygenase ◽  
Emerging Pollutant ◽  
P450 Enzyme

Abstract Background: The chloroacetamide herbicides pretilachlor is an emerging pollutant. Due to the large amount of use, its presence in the environment threatens human health. However, the molecular mechanism of pretilachlor degradation remains unknown. Results: Now, Rhodococcus sp. B2 was isolated from rice field and shown to degrade pretilachlor. The maximum pretilachlor degradation efficiency (86.1%) was observed at a culture time of 5 d, an initial substrate concentration 50 mg/L, pH 6.98, and 30.1°C. One novel metabolite N-hydroxyethyl-2-chloro-N-(2, 6-diethyl-phenyl)-acetamide was identified by gas chromatography-mass spectrometry (GC-MS). Draft genome comparison demonstrated that a 32,147-bp DNA fragment, harboring gene cluster (EthRABCDB2), was absent from the mutant strain TB2 which could not degrade pretilachlor. The Eth gene cluster, encodes an AraC/XylS family transcriptional regulator (EthRB2), a ferredoxin reductase (EthAB2), a cytochrome P450 monooxygenase (EthBB2), a ferredoxin (EthCB2) and a 10-kDa protein of unknown function (EthDB2). Complementation with EthABCDB2 and EthABDB2, but not EthABCB2 in strain TB2 restored its ability to degrade chloroacetamide herbicides. Subsequently, codon optimization of EthABCDB2 was performed, after which the optimized components were separately expressed in Escherichia coli, and purified using Ni-affinity chromatography. A mixture of EthABCDB2 or EthABDB2 but not EthABCB2 catalyzed the N-dealkoxymethylation of alachlor, acetochlor, butachlor, and propisochlor and O-dealkylation of pretilachlor, revealing that EthD acted as a ferredoxin in strain B2. EthABDB2 displayed maximal activity at 30 °C and pH 7.5. Conclusions: This is the first report of a P450 family oxygenase catalyzing the O-dealkylation and N-dealkoxymethylation of pretilachlor and propisochlor, respectively. And the results of the present study provide a microbial resource for the remediation of chloroacetamide herbicides-contaminated sites.

scholarly journals Molecular Modeling of Cytochrome P450 1A1 using the newly Crystallized Template structure of CYP1A2-Human

2021 ◽  
Vol 1 (1) ◽  
pp. 31-40
Author(s):  
Rehan Zafar ◽  
Fahed Parvaiz ◽  
Babar Aslam ◽  
Umar Niazi
Keyword(s):  
Cytochrome P450 ◽  
Active Site ◽  
Homology Model ◽  
Docking Studies ◽  
Cytochrome P450 Enzyme ◽  
Analysis Model ◽  
Cytochrome P450 1A1 ◽  
Entry Site ◽  
Monooxygenase Activity ◽  
Xenobiotic Detoxification

Cytochrome P450 enzyme family plays significant roles in carcinogenesis and xenobiotic detoxification. CYP1A1 is the P450 family 1 enzyme preferably expressed extrahepatically and participates extensively in monooxygenase activity which can either change the substrate to normal or carcinogenic metabolites, having the ability to initiate oncogenesis in lung and breast. Variegated structural properties evident in the prosites of available Cytochrome P450 (CYP) structures show versatility among CYP catalyzed reactions. In order to understand the CYP1A1 functions, hypothesized homology model has been constructed and characterization of the active site was performed by identifying important residues using docking studies and pharmacophore analysis. Model of CYP-1A1- Human has been constructed using the available crystal structure of CYP-1A2- Human. Active site and entry site of CYP-1A1 was found to be more compact than CYP1A2. Difference of wildtype CYP1A1 against its polymorphisms shows the role of mutations in active site architecture, which explains that the M2 and M4 mutations in CYP1A1 have no possible significant roles in the substrate binding and orientation for detoxification or carcinogenic activation. Different ligands including A- naphthoflavone (ANF), Ethoxyresorufin, Theophylline, Tamoxifen, Ethanol, Phenacetin and Hesperetin were docked and reconfirm the ligand specific wet lab studies. 

The role of cytochrome P450 enzyme genetic variants in cannabis hyperemesis syndrome—A case report

2021 ◽  
Author(s):  
Maxim Kuzin ◽  
Franziskos Xepapadakos ◽  
Isabel Scharrer ◽  
Marc Augsburger ◽  
Chin‐Bin Eap ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Case Report ◽  
Genetic Variants ◽  
Cytochrome P450 Enzyme ◽  
P450 Enzyme ◽  
Cannabis Hyperemesis Syndrome

Inter-species comparisons of hepatic cytochrome P450 enzyme levels in male ruminants

2003 ◽  
Vol 77 (10) ◽  
pp. 555-560 ◽  
Author(s):  
Miroslav Machala ◽  
Pavel Soucek ◽  
Jir� Neca ◽  
Robert Ulrich ◽  
Jir� Lamka ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Cytochrome P450 Enzyme ◽  
P450 Enzyme ◽  
Species Comparisons ◽  
Hepatic Cytochrome

scholarly journals Glutathione S-Transferases and Cytochrome P450 Enzyme Expression in Patients with Intracranial Tumors: Preliminary Report of 55 Patients

2018 ◽  
Vol 28 (1) ◽  
pp. 56-62
Author(s):  
Cahit Kural ◽  
Arzu Kaya Kocdogan ◽  
Gulcin Güler Şimşek ◽  
Serpil Oğuztüzün ◽  
Pınar Kaygın ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Brain Tissue ◽  
Pituitary Adenomas ◽  
Normal Brain ◽  
Cytochrome P450 Enzyme ◽  
Intracranial Tumors ◽  
Glutathione S Transferase ◽  
Normal Brain Tissue ◽  
Tissue Samples ◽  
P450 Enzyme

Objective: Intracranial tumors are one of the most frightening and difficult-to-treat tumor types. In addition to surgery, protocols such as chemotherapy and radiotherapy also take place in the treatment. Glutathione S-transferase (GST) and cytochrome P450 (CYP) enzymes are prominent drug-metabolizing enzymes in the human body. The aim of this study is to show the expression of GSTP1, GSTM1, CYP1A1, and CYP1B1 in different types of brain tumors and compare our results with those in the literature. Subjects and Methods: The expression of GSTP1, GSTM1, CYP1A1, and CYP1B1 was analyzed using immunostaining in 55 patients with intracranial tumors in 2016–2017. For GST and CYP expression in normal brain tissue, samples of a portion of surrounding normal brain tissue as well as a matched far neighbor of tumor tissue were used. The demographic features of the patients were documented and the expression results compared. Results: The mean age of the patients was 46.72 years; 29 patients were female and 26 were male. Fifty-seven specimens were obtained from 55 patients. Among them, meningioma was diagnosed in 12, metastases in 12, glioblastoma in 9, and pituitary adenoma in 5. The highest GSTP1, GSTM1, and CYP­1A1 expressions were observed in pituitary adenomas. The lowest GSTP1 expression was detected in glioblastomas and the lowest CYP1B1 expression in pituitary adenomas. Conclusion: GSTP1 and CYP expression is increased in intracranial tumors. These results should be confirmed with a larger series and different enzyme subtypes.

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