Cytochrome P450s Research Articles

Effects of Amino Acid Mutation in Cytochrome P450 (CYP96A146) of Descurainia sophia on the Metabolism and Resistance to Tribenuron-Methyl.

Cytochrome P450 monooxygenases (P450s) play important roles in herbicide resistance. In this study, there are four amino acid mutations (F39Y, H163Y, S203A, and V361E) between CYP96A146-S and CYP96A146-R, which were cloned, respectively, from susceptible (S) and tribenuron-methyl-resistant (TR) Descurainia sophia. The Arabidopsis expressing CYP96A146-S or CYP96A146-R showed resistance to tribenuron-methyl, carfentrazone-ethyl, and oxyfluorfen, while Arabidopsis transformed with CYP96A146-R or CYP96A146 with any two or three mutations of H163Y, S203A, or V361E exhibited significantly higher resistance to tribenuron-methyl than Arabidopsis expressing CYP96A146-S. The metabolic rates of tribenuron-methyl were significantly faster in Arabidopsis expressing CYP96A146-R than that with CYP96A146-S. The molecular dynamics simulation demonstrated that amino acid mutations did not affect the domain of the HEM ring, which could significantly enhance the volume of the catalytic pocket in P450 (CYP96A146), thereby increasing the collision rate between the catalytic pocket and tribenuron-methyl. Hence, the amino acid mutations may be one of the mechanisms underlying P450-mediated herbicide resistance.

Insecticide resistance in the field populations of the Asian tiger mosquito Aedes albopictus in Beijing: resistance status and associated detoxification genes

BackgroundAedes (Stegomyia) albopictus (Skuse) is an invasive and widespread mosquito species that can transmit dengue, chikungunya, yellow fever, and Zika viruses. Its control heavily relies on the use of insecticides. However, the efficacy of the insecticide-based intervention is threatened by the increasing development of resistance to available insecticides. Understanding the current status and potential mechanisms of insecticide resistance is an important prerequisite for devising strategies to maintain the sustainability of vector control programs. In this study, we investigated the current status and probable candidate detoxification genes associated with insecticide resistance in the Asian tiger mosquito in Beijing, the capital city of China.MethodsBioassays were conducted on three field populations of Ae. albopictus collected from urban communities in Beijing by exposure to diagnostic doses of permethrin, deltamethrin, malathion, and propoxur. Differentially expressed genes (DEGs) associated with insecticide resistance were screened by transcriptomic analysis using Illumina RNA sequencing data (RNA-seq) from 12 independent RNA libraries constructed from female strains of the three field populations and one susceptible strain.ResultsThe bioassay results indicated that all the three field populations were resistant to propoxur (carbamate), deltamethrin, and permethrin (pyrethroids), but susceptible to malathion (organophosphate). Eighteen (18) cytochrome P450s (P450s), five (5) glutathione S-transferases (GSTs), four (4) carboxy/cholinesterases (CCEs), eight (8) UDP-glycosyltransferases (UGTs), and three (3) ATP-binding cassette transporters (ABCs) were found to be significantly overexpressed in the three field populations relative to the susceptible strain via transcriptomic analysis.ConclusionThis study demonstrates that the Ae. albopictus field populations in Beijing exhibit multiple phenotypic resistance to commonly used pyrethroids and carbamate. The identification of a number of DEGs associated with insecticide resistance indicates that the mechanisms underlying resistance in field populations are complicated, and detoxifying enzymes may play important roles. The multiple resistance status detected in the three field populations suggests that resistance management strategies such as insecticide rotation and non-chemical-based measures should be implemented in order to sustain effective control of the disease vector and vector-borne diseases.

Epoxy metabolites of linoleic acid promote the development of breast cancer via orchestrating PLEC/NFκB1/CXCL9-mediated tumor growth and metastasis

Breast cancer (BC) is a common malignant tumor in women and requires a comprehensive understanding of its pathogenesis for the development of new therapeutic strategies. Polyunsaturated fatty acids (PUFAs) metabolism-driven inflammation is a causative factor in cancer development. However, the function of PUFAs′ metabolism in BC remains largely unknown. Here we report the role and underlying mechanism of epoxyoctadecenoic acids (EpOMEs), the metabolites of linoleic acid mediated by cytochrome P450 (CYP) monooxygenases, in promoting the development of BC, particularly triple-negative BC (TNBC). A metabolomics study identified that EpOMEs were significantly increased in the plasma of BC patients and MMTV-PyMT mice, which accounted for the upregulation of CYP2J2 in BC tumor tissues and tumor cells. Decreased EpOMEs by treatment of CYP monooxygenase inhibitors significantly alleviated tumor development in MMTV-PyMT mice. Treatment with EpOMEs and overexpression of CYP2J2 to increase EpOMEs in TNBC cells significantly promoted cellular proliferation, migration, tumor growth, and metastasis. Whereas knockdown of CYP2J2 to decrease EpOMEs inhibited tumorigenesis and lung metastasis of TNBC, which was reversed by EpOME administration. Transcriptomics and proteomics analyses revealed CXCL9 and PLEC were critical for EpOME-mediated promotion of TNBC. Knockdown of CXCL9 and PLEC inhibited TNBC progression and EpOME-mediated promotion of TNBC. Both overexpression of CYP2J2 and EpOME treatment upregulate PLEC, while PLEC upregulates NFκB1, which is a transcription regulator of CXCL9. This study extends the understanding of the function of PUFAs metabolism in BC development, providing potential therapeutic targets and dietary guidelines for patients with TNBC and other BCs.The illustration of the hypothetical mechanism CYP2J2/EpOMEs promotes the tumorigenesis and metastasis of TNBC via PLEC/NFKB1/CXCL9 signaling pathway.

The AREB transcription factor SaAREB6 promotes drought stress-induced santalol biosynthesis in sandalwood

Abstract Sandalwood (Santalum album), a culturally significant and economically valuable horticultural species, is renowned for its heartwood and essential oils enriched with sesquiterpene compounds such as santalol. Despite progress in elucidating the biosynthetic pathway of these valuable metabolites, the transcriptional regulation of this process, particularly under abiotic stress conditions, remains largely unexplored. Under drought conditions, we observed a marked increase in SaAREB6 expression, paralleled by elevated levels of santalols. Moreover, we identified SaCYP736A167, a cytochrome P450 monooxygenase gene, as a direct target of SaAREB6. Using electrophoretic mobility shift assays (EMSAs), microscale thermophoresis assays (MSTs) and dual luciferase assays (DLAs), we validated the precise and specific interaction of SaAREB6 with the promoter region of SaCYP736A167. This interaction leads to the upregulation of SaCYP736A167, which in turn catalyzes the final steps in the conversion of sesquiterpene precursors to santalols, thereby reinforcing the connection between SaAREB6 activity and increased santalol production during drought. Collectively, our work illuminates the previously uncharacterized role of SaAREB6 in orchestrating a transcriptional regulation that facilitates drought-induced santalol biosynthesis in sandalwood, presenting opportunities for genetic engineering strategies to improve heartwood and essential oil yields in this economically vital species.

From Starfish to Gibberellins: Biosynthesis and Regulation of Plant Hormones.

I grew up with laboratory glassware and microscopes as treasures from a young age. I was a member of the Chemistry Club in junior high school, and when I visited RIKEN with club members, I wished to become an organic chemist in the future. I received my doctoral degree through the study of the spawning inhibitor of starfish. I became a researcher at RIKEN and identified the chemical structure of a mating pheromone of a yeast. As a plant biochemist, I studied a cell-free system of gibberellins at the University of Göttingen and tried to identify the gibberellin biosynthetic pathways in plants and clone gibberellin biosynthetic enzyme genes to understand the light regulation of plant growth. I also worked on biosynthetic enzymes of abscisic acid, indole acetic acid, and brassinosteroids. I developed a special interest in the oxygenases of plant hormone biosynthesis, cytochrome P450 monooxygenases, 2-oxoglutartae-dependent dioxygenase, molybdenum cofactor-containing oxidase, and flavin-containing monooxygenase.

Discovery and Functional Identification of 2,3-Oxidosqualene Cyclases and Cytochrome P450s in Triterpenoid Metabolic Pathways of Actinidia eriantha.

Actinidia eriantha Benth, known as the "king of fruits", is rich in triterpenoid compounds, particularly ursane-type and oleanane-type triterpenic acids. These secondary metabolites have been widely applied in medicine, cosmetics, agriculture, and other fields. To date, key enzyme genes involved in triterpenoid metabolic pathways in A. eriantha remain unexplored. This study employed transcriptome sequencing analysis combined with synthetic biology approaches involving heterologous expression in yeast to identify crucial genes responsible for the biosynthesis of triterpenoid components in A. eriantha: Two 2,3-oxidosqualene cyclases (AeOSC2 and AeOSC3) were characterized to catalyze the formation of major triterpene scaffolds, α-amyrin [precursor of ursolic acid (UA)], β-amyrin [precursor of oleanolic acid (OA)], and ψ-taraxasterol, and two cytochrome P450s (AeCYP716A8 and AeCYP716A9) mediating three-step oxidation at the C-28 position of ursane-type and oleanane-type triterpene scaffolds to form UA, OA, and intermediate oxidation products. We successfully reconstructed the biosynthetic pathway of ursane- and oleanane-type triterpenoids from A. eriantha in a heterologous yeast host and elucidated the two-step enzymatic reactions involved in triterpenoid biosynthesis. These findings lay the foundation for further understanding the biosynthesis of key active components in A. eriantha.

Two detoxification enzyme genes, CYP6DA2 and CarFE4, mediate the susceptibility to afidopyropen in Semiaphis heraclei

IntroductionSemiaphis heraclei is an important economic pest affecting Caprifoliaceae and Apiaceae plants, and chemical control is still the main effective control method in the field. Afidopyropen is a new type of pyridine cyclopropyl insecticide, which can effectively control piercing-sucking mouthparts pests and is suitable for pest resistance management. However, the detoxification mechanism of S. heraclei to afidopyropen is still poorly cleared.MethodsThe insecticidal activity of afidopyropen against S. heraclei and the enzyme activity assay and synergism bioassay were evaluated. The detoxification enzyme genes were obtained by transcriptome and validated by quantitative real-time PCR (RT-qPCR). Furthermore, RNA interference was used to study the functions of detoxification enzyme genes.ResultsThe activities of cytochrome P450 monooxygenases (P450s) and carboxylesterases (CarEs) were significantly increased under afidopyropen treatment. The toxicity of afidopyropen against S. heraclei was significantly increased after application the inhibitors of piperonyl butoxide and triphenyl phosphate. Sixteen P450 genes and three CarE genes were identified in the transcriptome of S. heraclei. The RT-qPCR results showed that eleven P450 genes and two CarE genes were significantly upregulated under afidopyropen treatment, and the expression of CYP6DA2 and CarFE4 was upregulated by more than 2.5 times. The expression pattern of CYP6DA2 and CarFE4 was further analyzed in different developmental stages of S. heraclei and knockdown of CYP6DA2 and CarFE4 significantly increased the susceptibility of S. heraclei to afidopyropen.ConclusionThe results of this study uncover the key functions of CYP6DA2 and CarFE4 in the detoxification mechanism of S. heraclei to afidopyropen, and provide a theoretical basis for the scientific use of afidopyropen in the field.

Model-Based Dose Selection of a Sphingosine-1-Phosphate Modulator, Etrasimod, in Patients with Various Degrees of Hepatic Impairment

Background/Objectives: Etrasimod is a newly FDA-approved Sphingosine-1-Phosphate modulator indicated for moderate and severe ulcerative colitis. It is extensively metabolized in the liver via the cytochrome P450 system and may accumulate markedly in patients with hepatic dysfunction, exposing them to toxicity. The aim of the current study is to utilize a physiologically-based pharmacokinetic modeling approach to evaluate the impact of hepatic impairment on the pharmacokinetic behavior of etrasimod and to appropriately select dosage regimens for patients with chronic liver disease; Methods: PK-Sim was used to develop the etrasimod PBPK model, which was verified using clinical data from healthy subjects and subsequently adapted to reflect the physiological changes associated with varying degrees of hepatic dysfunction; Results: Simulations indicated that hepatic clearance of etrasimod is clearly reduced in patients with Child–Pugh B and C liver impairment. Based on these findings, dosing adjustments were proposed to achieve therapeutic exposures equivalent to those in individuals with normal liver function. In the Child–Pugh B and C population groups, 75% and 62.5%, respectively, of the standard dose were enough to have comparable exposure to the healthy population. These adjusted dosages aim to mitigate the risk of drug toxicity while maintaining efficacy; Conclusions: The PBPK model provides a robust framework for individualizing drug therapy in patients with hepatic impairment, ensuring safer and more effective treatment outcomes. Further clinical studies are warranted to verify these dosing recommendations and to refine the model for broader clinical applications.

Metabolomic and transcriptomic analysis of Panax Notoginseng root: Mechanistic insights into the effects of flower bud removal on yield and phytochemical composition

The roots of Panax notoginseng (Burk.) F. H. Chen are used to treat organ damage and cardiovascular diseases in several East Asian countries. The removal of floral buds before inflorescence formation is widely employed to increase rhizome yield in the cultivation of Panax notoginseng. However, the molecular mechanisms regulating the growth and the accumulation of secondary metabolites remain unclear. We find that after the removal of flower buds, the root biomass accumulated in three-year-old Panax notoginseng after growing for four months increased by 27 %. Comparative transcriptomic and metabolomic analyses revealed that disbudding promoted the conversion of small molecule sugars to polysaccharides by regulating the expression of genes in the glycogen metabolism (GLG) family. It facilitated the accumulation of metabolites in the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway by upregulating the expression of isoprenoid synthase (ISP) family genes and inhibited the conversion from the tricarboxylic acid (TCA) cycle to the mevalonate (MVA) pathway by downregulating the expression of acetyl-CoA acetyltransferase (ACAT) family genes. At the same time, the total content of triterpene saponins, main active ingredients, basically remained unchanged. However, the content of intermediate saponins increased after the upregulation of cytochrome P450 monooxygenase (CYP) transcripts, while that of downstream saponins decreased after the downregulation of UDP-glucuronosyltransferase (UGT) transcripts. Additionally, scanning electron microscopy (SEM) observations of root cross-sections indicated that the disbudding potentially promoted the development of the primary root cortex and epidermis by regulating auxin-cytokinin (AUX-CTK) signaling, thus alleviating xylem cavitation. This study confirms the removal of flower buds as an effective way to enhance yield of Panax notoginseng, offering new insights into the basic mechanisms of growth regulation during plant reproductive development and providing valuable reference for studying metabolite accumulation patterns in Panax notoginseng and other medicinal plants in the Panax genus.

Multi-omics analysis of the toxic effects on gill tissues of crucian carp (Carassius auratus) from chronic exposure to environmentally relevant concentrations of Di(2-ethylhexyl) phthalate (DEHP)

The pervasive use of the plasticizer di(2-ethylhexyl) phthalate (DEHP) poses potential risks to global aquatic ecosystems. This study systematically evaluated the adverse effects of chronic exposure to environmentally relevant concentrations of DEHP on gill tissues of crucian carp, utilizing histological examination, metabolomic, and transcriptomic analysis. The results demonstrated that DEHP induced significant histopathological alterations in gill tissues, with significant enrichment observed in multiple pathways associated with amino acid, hormone, lipid, and xenobiotic metabolism. Metabonomics-transcriptomics analyses indicated that DEHP-induced significantly over-activation of cytochrome P450 1B1-like (p < 0.001) and cytochrome P450 3A30-like (p < 0.05) via the nuclear xenobiotic receptors pathway was a key factor contributing to the disruption of tryptophan metabolism and steroid hormone biosynthesis, as well as inducing circadian rhythm disruption. Moreover, circadian rhythm disruption further exacerbated the imbalance of cytochrome P450 (CYP450) enzyme system as well as linoleic acid, arachidonic acid, sphingolipid, and glycerophospholipid metabolism. Overall, the feedback regulation between the CYP450 enzyme system and circadian rhythms emerged as the primary mechanism underlying DEHP-induced metabolic and transcriptional disruptions, ultimately resulting in gill toxicity. This study not only enriched the toxic effects on aquatic organisms of chronic exposure to DEHP, but provided potential biomarkers for the environmental risk assessment of DEHP.

Compartmentalized Sesquiterpenoid Biosynthesis and Functionalization in the Chlamydomonas reinhardtii Plastid.

Terpenoids play key roles in cellular metabolism and can have specialized functions. Their heterologous production in microbial hosts offers an alternative to natural extraction. Here, we developed a subcellular engineering approach in the model green alga Chlamydomonas reinhardtii by targeting both sesquiterpenoid synthases and cytochrome P450s (CYPs) to the plastid, exploiting its photosynthetic electron transport chain to drive CYP-mediated oxidation without reductase partners. Nuclear-encoded sesquiterpenoid synthases were expressed with farnesyl pyrophosphate synthase fusions and targeted to the plastid, while CYPs were modified for soluble localization in the plastid stroma by removing transmembrane domains. The plastid environment supported hydroxylation, epoxidation, and oxidation reactions, with functionalization efficiencies reaching 80% of accumulated products. Carbon source availability influenced product ratios, revealing metabolic flexibility in the engineered pathways. Overall sesquiterpenoid yields ranged between 250-2500 µg L-1 under screening conditions, establishing proof-of-concept for using plastid biochemistry in complex terpenoid biosynthesis. Living two-phase terpenoid extractions with different perfluorinated solvents revealed variable performances based on sesquiterpenoid functionalization and solvent type. This work demonstrates that photosynthetic electron transport can drive CYP-mediated functionalization in engineered subcellular compartments. However, improvements in photobioreactor cultivation concepts will be required to facilitate the use of algal chassis for scaled production.

Characterization of acetolactate synthase genes and resistance mechanisms of multiple herbicide resistant Lolium multiflorum

Combining imidazolinone-tolerant wheat with imazamox presents an effective solution to combat weed resistance. However, Lolium multiflorum, a troublesome resistant weed infesting wheat fields, may have developed resistance to imazamox, and the potential resistance mechanisms are intriguing. In this study, we explored the susceptibility of L. multiflorum to imazamox and investigated the resistance mechanisms, including the contribution of the target enzyme acetolactate synthase (ALS) to resistance and the presence of non-target-site resistance (NTSR). Eight L. multiflorum populations suspected of being resistant to imazamox were collected, and six populations exhibited resistance, ranging from 2.45-fold to 16.32-fold. The LmALS1 gene from susceptible population D3 plants and multiple copies of the LmALS gene (LmALS1, LmALS2, LmALS2α, LmALS3, LmALS3α, LmALS3β) from resistant populations D5 and D8 plants were separately amplified. Two mutations (Pro/Gln197 to Thr, Trp574 to Leu) were found in LmALS1 in the resistant populations. Compared to D3, LmALS1 was overexpressed in D5 but not in D8. The presence of LmALS1 mutants (LmALS1-Thr197 and LmALS1- Leu574), along with LmALS2, LmALS3, and their subunits, contribute to the resistance phenotype by increasing bonding energies, weakening hydrogen bonds, or decreasing protein binding pocket volumes and surface area. Additionally, D5 and D8 populations exhibited multiple resistance (>40-fold) to three other ALS inhibitors: pyroxsulam, flucarbazone-sodium, and mesosulfuron-methyl. Pre-treatment with malathion and 4-chloro-7-nitrobenzoxadiazole (cytochrome P450 monooxygenase and glutathione S-transferase inhibitors respectively) reversed the resistance of the D8 population and partially reversed the resistance of the D5 population to imazamox. This study characterizes ALS genes and extends our knowledge into the ALS resistance mechanisms involved in L. multiflorum. It also deepens our understanding of the complex diversification resistance mechanisms, thereby facilitating advances in weed resistance management strategies in wheat fields.

Deciphering the biodegradation mechanism of 3,6-dichlorocarbazole using a novel isolate for its implication in bioaugmentation

This study reported a newly isolated strain, Streptomyces sp. FW-32, which showed good ability to degrade 3,6-dichlorocarbazole (3,6-DCCZ). The maximum degradation efficiency of 3,6-DCCZ reached 86.78 % within 5 d at pH of 7.6, glucose concentration of 5.0 g/L, temperature of 33 °C, and 3,6-DCCZ initial concentration of 1.0 mg/L. In total, five biodegradation products were detected, indicating that 3,6-DCCZ was mainly transformed via dechlorination, oxidation, and aromatic ring cleavage. Proteomic analysis suggested that haloacid dehalogenase, glutathione S-transferase family protein, cytochrome P450s, pentachlorophenol monooxygenase, hydroxylase, and several dioxygenases were involved in 3,6-DCCZ biotransformation. Glutathione peroxidase, catalases, catalase − peroxidase, thioredoxin, thioredoxin reductase, and thioredoxin-dependent peroxiredoxin were induced to resist 3,6-DCCZ stress. Molecular docking revealed that 3,6-DCCZ exhibited an affinity for the potential proteins involved in 3,6-DCCZ biodegradation and stress response. Furthermore, the biotreatment of 3,6-DCCZ with the strain FW-32 might have contributed to the reduction in the aquatic toxicity of 3,6-DCCZ. The strain FW-32 efficiently synergized with certain indigenous microorganisms, consequently accelerating the removal of 3,6-DCCZ from the water–sediment system. Overall, these findings shed a new light on the mechanisms underlying the microbial transformation of 3,6-DCCZ, suggesting that Streptomyces sp. FW-32 exhibits considerable potential for the bioaugmentation of 3,6-DCCZ-contaminated aquatic environments.

Recent Advances and Challenges in the Production of Hydroxylated Natural Products Using Microorganisms

Hydroxylation reaction is a significant source of structural diversity in natural products (NPs), playing a crucial role in improving the bioactivity, solubility, and stability of natural product molecules. This review summarizes the latest research progress in the field of natural product hydroxylation, focusing on several key hydroxylases involved in the biosynthesis of NPs, including cytochrome P450 monooxygenases, α-ketoglutarate-dependent hydroxylases, and flavin-dependent monooxygenases. These enzymes achieve selective hydroxylation modification of various NPs, such as terpenoids, flavonoids, and steroids, through different catalytic mechanisms. This review systematically summarizes the recent advances on the hydroxylation of NPs, such as amino acids, steroids, terpenoids, lipids, and phenylpropanoids, demonstrating the potential of synthetic biology strategies in constructing artificial biosynthetic pathways and producing hydroxylated natural product derivatives. Through metabolic engineering, enzyme engineering, genetic engineering, and synthetic biology combined with artificial intelligence-assisted technologies, a series of engineered strains have been successfully constructed for the efficient production of hydroxylated NPs and their derivatives, achieving efficient synthesis of hydroxylated NPs. This has provided new avenues for drug development, functional food, and biomaterial production and has also offered new ideas for the industrial production of these compounds. In the future, integrating artificial synthetic pathway design, enzyme directed evolution, dynamic regulation, and artificial intelligence technology is expected to further expand the application of enzyme-catalyzed hydroxylation reactions in the green synthesis of complex NPs, promoting research on natural product hydroxylation to new heights.

Oxidation of four monoterpenoid indole alkaloid classes by three cytochrome P450 monooxygenases from Tabernaemontana litoralis.

Cytochrome P450 monooxygenases (CYPs) are well known for their ability to catalyze diverse oxidation reactions, playing a significant role in the biosynthesis of various natural products. In the realm of monoterpenoid indole alkaloids (MIAs), one of the largest groups of alkaloids in nature, CYPs are integral to reactions such as hydroxylation, epoxidation, ring opening, ring rearrangement, and aromatization, contributing to the extensive diversification of these compounds. In this study, we investigate the transcriptome, metabolome, and MIA biosynthesis in Tabernaemontana litoralis (milky way tree), a prolific producer of rare pseudoaspidosperma-type MIAs. Alongside known pseudoaspidosperma biosynthetic genes, we identify and characterize three new CYPs that facilitate regio- and stereospecific oxidation of four MIA skeletons: iboga, aspidosperma, pseudoaspidosperma, and quebrachamine. Notably, the tabersonine 14,15-β-epoxidase catalyzes the formation of pachysiphine, the stereoisomer of 14,15-α-epoxytabersonine (lochnericine) found in Catharanthus roseus (Madagascar periwinkle) roots. The pseudovincadifformine 18-hydroxylase is the first CYP identified to modify a pseudoaspidosperma skeleton. Additionally, we demonstrate that the enzyme responsible for C10-hydroxylation of the iboga MIA coronaridine also catalyzes C10-hydroxylation of voaphylline, which bears a quebrachamine skeleton. With the discovery of a new MIA, 11-hydroxypseudovincadifformine, this study provides a comprehensive understanding of MIA biosynthesis and diversification in T. litoralis, highlighting its potential for further exploration.

Knockdown of CYP6SZ3 and CYP6AEL1 genes increases the susceptibility of Lasioderma serricorne to ethyl formate and benzothiazole.

Insect cytochrome P450 monooxygenases (CYPs) play crucial roles in the metabolic detoxification of insecticides. Ethyl formate and benzothiazole have recently regained popularity as fumigants due to rising resistance to phosphine in the stored-product pests. However, the mechanisms underlying tolerance to these two fumigants in Lasioderma serricorne, a major global insect pest of stored products, remain poorly understood. In this study, two CYP genes, named CYP6SZ3 and CYP6AEL1, were identified from L. serricorne, belonging to the CYP6 family and containing five conserved domains characteristic of CYP proteins. Spatiotemporal expression analysis revealed that both genes were predominantly expressed in the larval stage and showed the highest expression in the foregut. Upon exposure to ethyl formate and benzothiazole, both genes were upregulated, with significantly increased transcription levels following treatment. RNA interference-mediated silencing of CYP6SZ3 and CYP6AEL1 led to increased susceptibility and significantly higher mortality of L. serricorne when exposed to these fumigants. Homology modeling and molecular docking analyses showed stable binding of these fumigants to CYP6SZ3 and CYP6AEL1 proteins, with binding free energies from -26.88 to -94.68 kcal mol-1. These findings suggest that the induction of CYP6SZ3 and CYP6AEL1 is likely involved in the detoxification of ethyl formate and benzothiazole in L. serricorne.

Retraction of "Set of Cytochrome P450s Cooperatively Catalyzes the Synthesis of a Highly Oxidized and Rearranged Diterpene-Class Sordarinane Architecture".

Retraction of "Set of Cytochrome P450s Cooperatively Catalyzes the Synthesis of a Highly Oxidized and Rearranged Diterpene-Class Sordarinane Architecture".

Tuning architectural organization of eukaryotic P450 system to boost bioproduction in Escherichia coli

Eukaryotic cytochrome P450 enzymes, generally colocalizing with their redox partner cytochrome P450 reductase (CPR) on the cytoplasmic surface of organelle membranes, often perform poorly in prokaryotic cells, whether expressed with CPR as a tandem chimera or free-floating individuals, causing a low titer of heterologous chemicals. To improve their biosynthetic performance in Escherichia coli, here, we architecturally design self-assembled alternatives of eukaryotic P450 system using reconstructed P450 and CPR, and create a set of N-termini-bridged P450-CPR heterodimers as the counterparts of eukaryotic P450 system with N-terminus-guided colocalization. The covalent counterparts show superior and robust biosynthetic performance, and the N-termini-bridged architecture is validated to improve the biosynthetic performance of both plant and human P450 systems. Furthermore, the architectural configuration of protein assemblies has an inherent effect on the biosynthetic performance of N-termini-bridged P450-CPR heterodimers. The results suggest that spatial architecture-guided protein assembly could serve as an efficient strategy for improving the biosynthetic performance of protein complexes, particularly those related to eukaryotic membranes, in prokaryotic and even eukaryotic hosts.

The Metabolic Characteristics and Bioavailability of Resveratrol Based on Metabolic Enzymes.

The natural polyphenol resveratrol (RV) has garnered fame for its extensive pharmacological properties. Although clinical studies have shown some positive results, many contradictory outcomes remain. An important obstacle to the development of therapeutic applications for RV is its low bioavailability in vivo. This may be partially attributed to biotransformation mediated by phase I and II enzymes, such as cytochrome P450s, UDP-glucuronosyltransferases, and sulfotransferases. To date, more than 20 different types of metabolites have been detected after catalysis by these enzymes. Notably, RV and some of its metabolites serve as substrates for these enzymes. Conversely, RV can directly regulate the expression or activity of these enzymes. Given the increasing number of studies investigating the bioactivity of RV, this review summarizes its physicochemical and pharmacokinetic characteristics and describes the metabolism of RV and the bioactivities of its metabolites, with emphasis on the interaction between RV and its related metabolic enzymes. In addition to hepatic metabolism, the crucial roles of RV metabolism in multiple other tissues and organs cannot be overlooked, and they reveal the relationship between RV metabolism and its biological potential.

PlZAT10 binds to the ABA catabolism gene PlCYP707A2 promoter to mediate seed dormancy release in Paeonia lactiflora.

PlZAT10-PlCYP707A2 module promotes Paeonia lactiflora seeds germination. The herbaceous peony (Paeonia lactiflora) seeds exhibit double dormancy in the epicotyl and hypocotyl, which significantly inhibits the process of cultivation and breeding of new varieties. Nevertheless, the molecular mechanism underlying seed dormancy release in P. lactiflora remains to be fully identified.In this current study, we analyzed differentially expressed genes based on transcriptome data and selected the abscisic acid catabolic gene PlCYP707A2 for further investigation. The conserved domain of the protein indicated that PlCYP707A2 possessed a cytochrome P450 monooxygenase domain. Subcellular localization indicated that PlCYP707A2 was localized on the cytoplasm and cell membrane. Overexpression of PlCYP707A2 in P. lactiflora seeds decreased ABA contents and promoted seeds germination. The silencing of PlCYP707A2 resulted in seed dormancy and an alteration in the content of ABA. Moreover, yeast one-hybrid, electrophoretic mobility shift and dual-luciferase reporter assay revealed that PlZAT10 bound to the promoter of PlCYP707A2. In conclusion, the results demonstrated the mechanism of the PlZAT10-PlCYP707A2 module in regulating the dormancy release of P. lactiflora seeds, enriching relevant theories on seed dormancy and having significant implications for the herbaceous peony industry developing.