Evaluation of Recombinant Cytochromes P450 Activity in Metabolic Pathways

Bertrand Rochat

doi:10.1124/dmd.31.1.145

Five genetic polymorphisms of cytochrome P450 enzymes in the Czech non-Roma and Czech Roma population samples

Drug Metabolism and Personalized Therapy ◽

10.1515/dmdi-2020-0103 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Lucie Dlouhá ◽

Věra Adámková ◽

Lenka Šedová ◽

Věra Olišarová ◽

Jaroslav A. Hubáček ◽

...

Keyword(s):

Cytochrome P450 ◽

Genetic Polymorphisms ◽

Metabolic Pathways ◽

Cytochromes P450 ◽

Taqman Assay ◽

Cytochrome P450 Enzymes ◽

Roma Population ◽

Czech Population ◽

Genotype Frequencies ◽

Pcr Rflp

AbstractObjectivesCytochromes P450 play a role in human drugs metabolic pathways and their genes are among the most variable in humans. The aim of this study was to analyze genotype frequencies of five common polymorphisms of cytochromes P450 in Roma/Gypsy and Czech (non-Roma) population samples with Czech origin.MethodsRoma/Gypsy (n=302) and Czech subjects (n=298) were genotyped for CYP1A2 (rs762551), CYP2A6 (rs4105144), CYP2B6 (rs3745274) and CYP2D6 (rs3892097; rs1065852) polymorphisms using PCR-RFLP or Taqman assay.ResultsWe found significant allelic/genotype differences between ethnics in three genes. For rs3745274 polymorphism, there was increased frequency of T allele carriers in Roma in comparison with Czech population (53.1 vs. 43.7%; p=0.02). For rs4105144 (CYP2A6) there was higher frequency of T allele carriers in Roma in comparison with Czech population (68.7 vs. 49.8%; p<0.0001). For rs3892097 (CYP2D6) there was more carriers of the A allele between Roma in comparison with Czech population (39.2 vs. 38.2%; p=0.048). Genotype/allelic frequencies of CYP2D6 (rs1065852) and CYP1A2 (rs762551) variants did not significantly differ between the ethnics.ConclusionsThere were significant differences in allelic/genotype frequencies of some, but not all cytochromes P450 polymorphisms between the Czech Roma/Gypsies and Czech non-Roma subjects.

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Impact of SARS‐CoV‐2 infection (COVID‐19) on cytochromes P450 activity assessed by the Geneva cocktail

Clinical Pharmacology & Therapeutics ◽

10.1002/cpt.2412 ◽

2021 ◽

Author(s):

Camille Lenoir ◽

Jean Terrier ◽

Yvonne Gloor ◽

François Curtin ◽

Victoria Rollason ◽

...

Keyword(s):

Cytochromes P450 ◽

P450 Activity

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Interactions between dietary flavonoids and 2,3,7,8-tetrachlorodibenzo-p-dioxin on cytochromes P450 activity in a human intestinal model (Caco-2 cells)

Toxicology Letters ◽

10.1016/j.toxlet.2007.05.263 ◽

2007 ◽

Vol 172 ◽

pp. S95-S96

Author(s):

Thérèse Sergent ◽

Isabelle Dupont ◽

Luc Pussemier ◽

Yvan Larondelle ◽

Yves-Jacques Schneider

Keyword(s):

Cytochromes P450 ◽

Tetrachlorodibenzo P Dioxin ◽

Dietary Flavonoids ◽

P450 Activity

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Impact of acute inflammation on cytochromes P450 activity assessed by the Geneva Cocktail.

Clinical Pharmacology & Therapeutics ◽

10.1002/cpt.2146 ◽

2020 ◽

Author(s):

Camille Lenoir ◽

Youssef Daali ◽

Victoria Rollason ◽

François Curtin ◽

Yvonne Gloor ◽

...

Keyword(s):

Acute Inflammation ◽

Cytochromes P450 ◽

P450 Activity

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Personalized pharmacotherapy of arterial hypertension patients with musculoskeletal system diseases based on pharmacogenetic aspects

Pharmacogenetics and Pharmacogenomics ◽

10.37489/2588-0527-2021-1-24-32 ◽

2021 ◽

pp. 24-32

Author(s):

K. M. Muratov ◽

I. V. Stuk ◽

N. I. Lapudus

Keyword(s):

Current Issue ◽

Antihypertensive Drugs ◽

Musculoskeletal Diseases ◽

Cytochromes P450 ◽

Term Therapy ◽

Inflammatory Drugs ◽

Long Term Therapy ◽

A Current ◽

P450 Activity

Pharmacotherapy in patients with comorbidity is a current issue for clinical practice. Combination of hypertension and musculoskeletal diseases can be found in 40% of outpatients, which requires simultaneous administration of different drugs. The main mechanisms of drug interactions are associated with pharmacokinetics or pharmacodynamics alterations. It has been proven that changes in drugs pharmacokinetics can be due to cytochromes P450 activity. The main symptom of musculoskeletal diseases is chronic pain, which requires long-term therapy with non-steroidal anti-inflammatory drugs (NSAIDs). The 2C19 isoenzyme takes part in metabolism of some NSAIDs. Losartan, the inhibitor of renin-angiotensinaldosterone system (RAAS), is also metabolized by the 2C9 isoenzyme and is quite often prescribed to outpatients to treat hypertension. Hence, an influence of genetic factors on efficacy and safety of antihypertensive drugs and NSAIDs combinations requires further studies.

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Evaluation of P450 monooxygenase activity in lyophilized recombinant E. coli cells compared to resting cells

AMB Express ◽

10.1186/s13568-021-01319-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Thomas Hilberath ◽

Alessandra Raffaele ◽

Leonie M. Windeln ◽

Vlada B. Urlacher

Keyword(s):

Cytochromes P450 ◽

Rhodococcus Erythropolis ◽

Resting Cells ◽

P450 Monooxygenase ◽

Monooxygenase Activity ◽

Whole Cells ◽

E Coli ◽

Redox Partner ◽

Starting Point ◽

P450 Activity

AbstractCytochromes P450 catalyze oxidation of chemically diverse compounds and thus offer great potential for biocatalysis. Due to the complexity of these enzymes, their dependency of nicotinamide cofactors and redox partner proteins, recombinant microbial whole cells appear most appropriate for effective P450-mediated biocatalysis. However, some drawbacks exist that require individual solutions also when P450 whole-cell catalysts are used. Herein, we compared wet resting cells and lyophilized cells of recombinant E. coli regarding P450-catalyzed oxidation and found out that lyophilized cells are well-appropriate as P450-biocatalysts. E. coli harboring CYP105D from Streptomyces platensis DSM 40041 was used as model enzyme and testosterone as model substrate. Conversion was first enhanced by optimized handling of resting cells. Co-expression of the alcohol dehydrogenase from Rhodococcus erythropolis for cofactor regeneration did not affect P450 activity of wet resting cells (46% conversion) but was crucial to obtain sufficient P450 activity with lyophilized cells reaching a conversion of 72% under the same conditions. The use of recombinant lyophilized E. coli cells for P450 mediated oxidations is a promising starting point towards broader application of these enzymes.

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