Reductive Cytochrome P450 Reactions and Their Potential Role in Bioremediation

Behrendorff, James B. Y. H.

doi:10.3389/fmicb.2021.649273

Cited by 31 publications

(13 citation statements)

References 106 publications

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“…The enzyme co-factors or initial acceptors like CPR can accept electrons directly from the electrode that can act as an electron source for P450s. [28][29][30][31] In bioelectrocatalysis, enzymes can mimic their natural pathway for Cyt P450-catalyzed oxidations on electrodes as demonstrated in the thin LbL films made from polyions, and Cyt P450 CPR microsomes containing pure Cyt P450. 131 These stable films consisted of thin layers of the polyions poly(diallyldimethylammonium chloride) (PDDA) and PSS as six alternating layers with CYP2A1 microsomes (containing CPR), denoted as PDDA/PSS(/CYP1A2/CPR) 6 films on PG electrodes.…”

Section: Activation Of Enzymes In Electrochemistrymentioning

confidence: 99%

“…7,[14][15][16][17][18][19] Cyt P450s are biocatalysts for C-H hydroxylation, N-and S-oxidation, O-, N-and S-dealkylation, C-C bond cleavage, CQC double bond epoxidation, aromatic coupling, as well as more unusual reactions. 20,21 They are effective for synthesis of drugs, drug metabolites, [22][23][24][25][26][27] bioremediation, [28][29][30] and development of biosensors. [31][32][33] Cyt P450s as monooxygenases incorporate one oxygen atom site-selectively into a reactive substrate.…”

Section: Introductionmentioning

confidence: 99%

“…7,14–19 Cyt P450s are biocatalysts for C–H hydroxylation, N- and S-oxidation, O-, N- and S-dealkylation, C–C bond cleavage, CC double bond epoxidation, aromatic coupling, as well as more unusual reactions. 20,21 They are effective for synthesis of drugs, drug metabolites, 22–27 bioremediation, 28–30 and development of biosensors. 31–33…”

Section: Introductionmentioning

confidence: 99%

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Electrochemical transformations catalyzed by cytochrome P450s and peroxidases

Kumar

Rusling

2023

Chem. Soc. Rev.

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Section: Activation Of Enzymes In Electrochemistrymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Electrochemical transformations catalyzed by cytochrome P450s and peroxidases

Kumar

Rusling

2023

Chem. Soc. Rev.

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“…As such they are attractive compartments for introducing a variety of metabolic pathways, such as those dependent on oxyfunctionalization by cytochrome P450 enzymes ( Mellor et al., 2016 ). Because of the widespread occurrence of P450s throughout all kingdoms of life, their prominent roles in specialized metabolism and ability to catalyze remarkably diverse chemical transformations ( Podust and Sherman, 2012 ; Liu et al., 2020 ; Behrendorff, 2021 ), they are important enzymes in the metabolic engineering toolbox. Most P450s in eukaryotes localize to the endoplasmic reticulum (ER) where they are anchored by a single N-terminal transmembrane domain.…”

Section: Introductionmentioning

confidence: 99%

Exploiting photosynthesis-driven P450 activity to produce indican in tobacco chloroplasts

et al. 2023

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Photosynthetic organelles offer attractive features for engineering small molecule bioproduction by their ability to convert solar energy into chemical energy required for metabolism. The possibility to couple biochemical production directly to photosynthetic assimilation as a source of energy and substrates has intrigued metabolic engineers. Specifically, the chemical diversity found in plants often relies on cytochrome P450-mediated hydroxylations that depend on reductant supply for catalysis and which often lead to metabolic bottlenecks for heterologous production of complex molecules. By directing P450 enzymes to plant chloroplasts one can elegantly deal with such redox prerequisites. In this study, we explore the capacity of the plant photosynthetic machinery to drive P450-dependent formation of the indigo precursor indoxyl-β-D-glucoside (indican) by targeting an engineered indican biosynthetic pathway to tobacco (Nicotiana benthamiana) chloroplasts. We show that both native and engineered variants belonging to the human CYP2 family are catalytically active in chloroplasts when driven by photosynthetic reducing power and optimize construct designs to improve productivity. However, while increasing supply of tryptophan leads to an increase in indole accumulation, it does not improve indican productivity, suggesting that P450 activity limits overall productivity. Co-expression of different redox partners also does not improve productivity, indicating that supply of reducing power is not a bottleneck. Finally, in vitro kinetic measurements showed that the different redox partners were efficiently reduced by photosystem I but plant ferredoxin provided the highest light-dependent P450 activity. This study demonstrates the inherent ability of photosynthesis to support P450-dependent metabolic pathways. Plants and photosynthetic microbes are therefore uniquely suited for engineering P450-dependent metabolic pathways regardless of enzyme origin. Our findings have implications for metabolic engineering in photosynthetic hosts for production of high-value chemicals or drug metabolites for pharmacological studies.

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“…The genome sequencing of these mycobacterial species revealed a large number of operons, which are involved in the degradation of lipids. This includes a number of genes encoding cytochrome P450 enzymes (CYPs for example: CYP51, CYP124, CYP125, and CYP142), which have been proposed to be involved in steroid metabolism. , These P450 monooxygenase enzymes contain a heme-thiolate cofactor and perform the catalytic insertion of an oxygen atom into a C–H bond and are capable of supporting other reactions. − …”

mentioning

confidence: 99%

The Structures of the Steroid Binding CYP142 Cytochrome P450 Enzymes from Mycobacterium ulcerans and Mycobacterium marinum

et al. 2022

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The steroid binding CYP142 cytochrome P450 enzymes of Mycobacterium species are involved in the metabolism of cholesterol and its derivatives. The equivalent enzyme from Mycobacterium ulcerans was studied to compare the degree of functional conservation between members of this CYP family. We compared substrate binding of the CYP142A3 enzymes of M. ulcerans and M. marinum and CYP142A1 from M. tuberculosis using UV–vis spectroscopy. The catalytic oxidation of cholesterol derivatives by all three enzymes was undertaken. Both CYP142A3 enzymes were structurally characterized by X-ray crystallography. The amino acid sequences of the CYP142A3 enzymes are more similar to CYP142A1 from M. tuberculosis than CYP142A2 from Mycolicibacterium smegmatis. Both CYP142A3 enzymes have substrate binding properties, which are more resemblant to CYP142A1 than CYP142A2. The cholest-4-en-3-one-bound X-ray crystal structure of both CYP142A3 enzymes were determined at a resolution of <1.8 Å, revealing the substrate binding mode at a high level of detail. The structures of the cholest-4-en-3-one binding CYP142 enzymes from M. ulcerans and M. marinum demonstrate how the steroid binds in the active site of these enzymes. They provide an explanation for the high selectivity of the enzyme for terminal methyl C–H bond oxidation to form 26-hydroxy derivatives. These enzymes in pathogenic Mycobacterium species are candidates for inhibition. The work here demonstrates that similar drug molecules could target these CYP142 enzymes from different species in order to combat Buruli ulcer or tuberculosis.

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Reductive Cytochrome P450 Reactions and Their Potential Role in Bioremediation

Cited by 31 publications

References 106 publications

Electrochemical transformations catalyzed by cytochrome P450s and peroxidases

Electrochemical transformations catalyzed by cytochrome P450s and peroxidases

Exploiting photosynthesis-driven P450 activity to produce indican in tobacco chloroplasts

The Structures of the Steroid Binding CYP142 Cytochrome P450 Enzymes from Mycobacterium ulcerans and Mycobacterium marinum

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