Cytochrome P450s Research Articles

Cloning and functional verification of Geraniol-10-Hydroxylase gene in Lonicera japonica.

Geraniol 10-hydroxylase (G10H) is a cytochrome P450 monooxygenase involved in regulation, which is involved in the biosynthesis of monoterpene. However, G10H is not characterized at the enzymatic mechanism and regulatory function in Lonicera japonica. A gene related to the biosynthesis of monoterpenoid, geraniol 10-hydroxylase, has been cloned from the medicinal plant Lonicera japonica. The gene, LjG10H, encodes a peptide of 498 amino acids with a predicted molecular weight of 54.45 kDa. LjG10H shares a homology of 72.93-83.90% with G10H from other plants. Phylogenetic analysis suggests that the protein encoded by this gene belongs to the cytochrome P450 monooxygenase family. Tissue-specific expression analysis revealed that LjG10H is most highly expressed in flowers. Through heterologous expression in E. coli, the LjG10H protein was purified and its catalytic activity was studied. The results show that the enzyme can catalyze the hydroxylation of geraniol to 10-hydroxygeraniol. Additionally, analysis of Lonicera japonica seedlings with silenced LjG10H revealed a reduction in monoterpenoid content. This study demonstrates that LjG10H plays an important role in the biosynthetic pathway of iridoids. This is the first article that ascribes G10H to be associated with the biosynthetic pathway of iridoid. This study provides a theoretical basis for the functional mechanism of LjG10H in regulating iridoid synthesis and provides a valuable resource for molecular breeding studies.

Investigating non-target site resistance to pyroxsulam in a glyphosate-resistant Lolium rigidum population.

Resistance to multiple herbicides is common in Lolium rigidum. Here, resistance to acetolactate synthase (ALS)- and susceptibilityto acetyl-CoA carboxylase (ACCase)-inhibiting herbicides was confirmed in a glyphosate-resistant L. rigidum population (NLR70) from Australia and the mechanisms of pyroxsulam resistance were examined. No ALS target-site mutations nor gene overexpression were detected. Cytochrome P450 monooxygenase (P450) and glutathione S-transferase (GST) inhibitors (indicators of some certain P450s or GSTs) did not significantly affect the resistance to pyroxsulam. Nevertheless, HPLC analysis showed that plants of the NLR70 population metabolized pyroxsulam faster than plants of the herbicide-susceptible population (SVLR1). RNA sequencing analysis and RT-qPCR validation confirmed that four P450s (CYP709B2, CYP72A14, CYP89A2, CYP94B3), one GT (UGT79), and one ABC transporter (ABCG41) genes were constitutively upregulated in NLR70 plants. This study demonstrates that the glyphosate-resistant L. rigidum population (NLR70) also exhibits resistance to pyroxsulam and identifies six candidate genes associated with non-target site resistance to pyroxsulam. © 2025 Society of Chemical Industry.

Discovering the compatibility of Coptidis Rhizoma and Gardeniae Fructus in attenuating hepatotoxicity through the association rules analysis of related formula.

Gardenia jasminoides J. Ellis (Gardeniae Fructus, GF) is a widely used herbal medicine in many prescriptions. However, inappropriate application of GF may induce hepatotoxicity, which greatly challenges its clinical application. In the field of Traditional Chinese Medicine, the principle of "Compatibility for Toxicity Attenuation" is a pivotal concept. So far, the basis and mechanisms of compatibility with GF remain unclear. We aimed to investigate the toxicity attenuating effects and potential mechanisms of compatibility of Coptis chinensis Franch (Coptidis Rhizoma, CR) and GF in hepatotoxicity. Association rules analysis was performed in 817 GF-related formulas, and 49 associated items were selected, among which CR and GF were ranked in the top two. Network pharmacology was used to elucidate the potential mechanisms of combination of CR and GF in mitigating hepatotoxicity. Nine potential hepatotoxic components of GF and 14 active components of CR were focused on, and 9 common targets for CR and GF were identified. Further GO and KEGG analyses showed that the toxicity attenuating effect of CR on GF-induced hepatotoxicity may be closely correlated with inflammatory response, response to hypoxia, cancer signaling pathways, PI3K-Akt and HIF-1 signaling pathway. The in vitro results indicated that the combined use of CR and GF increased the viability of HepG2 cells. Furthermore, the in vivo data demonstrated that CR inhibited GF-induced increase in serum ALT and AST levels and pathological changes in the livers of KM mice. Besides, CR reduced the accumulation of ROS and MDA, inhibited the release of TNF-α and IL-6, while elevated GSH level in the mouse liver tissues. Finally, the molecular docking results indicated that the active components of CR had strong binding affinity with the monooxygenase cytochrome P450 3A4 (CYP3A4). CR combination restored the expression of CYP3A4 in the liver tissues of mice challenged with GF. Co-administration with CR effectively reduced GF-induced hepatotoxicity through alleviating oxidative damage, inflammatory response and enhancing CYP3A4 expression.

Insect Cytochrome P450 Database: An Integrated Resource of Genetic Diversity, Evolution and Function.

Insects, the most numerous and diverse group of animal species on Earth, have important interactions with humans through providing resources, transmitting diseases and damaging agricultural cultivars. Cytochrome P450 monooxygenases (P450s) are one of the most important protein families in insects implicated in the endogenous metabolism and detoxification of xenobiotics, including allelochemicals, insecticides and environmental pollutants. To better understand the evolution and function of insect P450s and support the development and application of insecticides for pest control, an integrated bioinformatics platform is highly desirable. Here, we present the Insect Cytochrome P450 database (ICPD, http://www.insectp450.net/), which contains 66,477 P450s collected from public databases and predicted from the genomes of 682 insect species using a standardised bioinformatics protocol. Phylogenetic relationships between P450 genes are constructed for each species. The structures of all P450 proteins in the database are predicted using ESMFold, then visualised using WeView. Web services, such as BLAST, homogeneous modelling and molecular docking, are provided for determining the catalytic activities of P450 proteins. The ICPD will facilitate systematic investigations of the evolution and functions of the complete insect P450 complement, and represents a powerful tool for guiding insecticide design and application.

Crystal structure of cytochrome P450 NysL and the structural basis for stereo- and regio- selective oxidation of antifungal macrolides.

NysL, a cytochrome P450 monooxygenase from the Gram-positive bacterium Streptomyces noursei, catalyzes the C10 hydroxylation of 10-deoxynystain to nystatin A1, a clinically important antifungal. In this study, we present the 2.0 Å resolution crystal structure of NysL bound to nystatin A1. The structure of this complex provides key insights into the structural elements that dictate the regio- and stereo- selective oxidation of large 20-44-membered macrolide substrates. The closely related AmphL operates on a similar 38-member macrolide but oxidizes C8 rather than C10. This difference requires that the substrate for AmphL penetrate further into the active site relative to NysL. The depth of substrate penetration is controlled by interactions between an area of the substrate binding pocket deemed the "back-wall" and the hemiketal ring of the macrolide substrate.

The bitter truth: how insects cope with toxic plant alkaloids.

Plants are unique organisms that have developed ingenious strategies to cope with environmental challenges, such as herbivorous insects. One of these strategies is the synthesis of a vast array of chemical compounds, known as specialized metabolites, that serve many ecological functions. Among the most fascinating and diverse groups of specialized metabolites are the alkaloids, which are characterized by the presence of a nitrogen atom within a heterocyclic ring. While some have medicinal and recreational applications, others are highly unpalatable and/or toxic. The effects of alkaloids on both humans and insects can be very diverse, affecting their physiology and behavior. Insects that feed on alkaloid-containing plants have evolved various mechanisms to cope with the consequences of these toxins. These include sequestration, where insects store alkaloids in specialized tissues or organs, enzymatic detoxification through enzymes such as cytochrome P450 monooxygenases and glutathione S-transferases, and behavioral adaptations such as selective feeding. In this review, we explore the relationships between plant alkaloids and the evolutionary adaptations that enable insects to exploit alkaloid-rich plants as food sources and ecological niches minimizing the harmful effects of these natural compounds. We aim to provide a comprehensive and updated overview of this fascinating and complex ecological interaction.

In vitro Metabolism and Cytotoxicity of Parthenolide: The Complete Identification of the Major Oxidative Product and the Evaluation of Trypanocidal and Leishmanicidal Activities

Parthenolide (PTN) is a secondary metabolite of the plant Tanacetum parthenium (L.) Schulz Bip. and is considered the chemical marker of this species. This sesquiterpene lactone (germacrene skeleton) has been described as responsible for the biological activity of the leaf extract. In addition, several studies in the literature have demonstrated its antiparasitic and antineoplastic activity. However, there is a need for knowledge of other safety parameters, such as pharmacokinetic, pharmacodynamic and toxicity evaluations. Therefore, in this investigation, the in vitro metabolism of parthenolide was performed with rat liver microsomes and biomimetic metabolism (such as cytochrome P-450 system) using organometallic catalysts. The biomimetic procedure was validated since the major compound of biomimetic oxidative reaction with meta-chloroperbenzoic acid catalyzed by metalloporphyrin 5,10,15,20-tetrakis(pentafluorophenyl)-porphyrin iron(III) chloride was also observed with rat liver microsomes. Previous chemoenzymatic synthesis studies of PTN afforded the same epoxide formation, this major oxidative compound was isolated and fully characterized as (1R,10R)-epoxyparthenolide based on experimental and theoretical calculations of infrared (IR) and vibrational circular dichroism (VCD) data at the B3PW91/6-311G. Furthermore, for the first time, this metabolite was evaluated for trypanocidal and leishmanicidal activity. The mean inhibitory concentration (IC50) values were lower for PTN than its metabolite and showed significant cytotoxic effects for both parasites. This finding holds promise for addressing neglected diseases.

Cytochrome P450 induced formation of hydroxylated polybrominated diphenyl ether and dibenzo-p-dioxin derivates from brominated phenols.

Methoxylated polybrominated diphenyl ethers (MeO-BDEs) are a class of environmentally relevant halogenated natural products. The two most relevant isomers, 2'-MeO-BDE 68 and 6-MeO-BDE 47, were repeatedly detected at levels comparable with persistent organic pollutants in marine environmental and food samples. MeO-BDEs were suspected to be biosynthesized by bromoperoxidases through the merging of two bromophenol units, three of which (2,4-dibromophenol, 2,6-dibromophenol and 2,4,6-tribromophenol) are abundant in marine environments, followed by O-methylation to give MeO-BDEs. However, not all abundant MeO-BDEs can be explained by the direct coupling of bromophenols. Therefore, several bromophenols were incubated with CYP109B1 from Bacillus subtilis. Incubations of 2,4-dibromophenol with CYP109B1 resulted in both 2'-OH-BDE 68 and 6-OH-BDE 47. Mechanistic considerations indicated that both compounds emerged from an epoxidized intermediate. After subsequent methylation this leads to the two most relevant MeO-BDEs in marine environments. By contrast, incubations with 2,6-dibromophenol and 2,4,6-tribromophenol did not form any OH-BDEs but tri- or tetrabrominated dibenzo-p-dioxins (PBDDs), i.e. another known class of environmentally relevant halogenated natural products whose toxicity has shown to be similar to their chlorinated analogues. SYNOPSIS: Treatment of bromophenols with cytochrome P450 monooxygenase CYP109B1 led to the formation of hydroxylated polybrominated diphenyl ethers and polybrominated dibenzo-p-dioxins.

Inheritance mode and metabolic mechanism of bromopropylate resistance in a field strain of Panonychus citri (Acari: Tetranychidae)

ABSTRACT Panonychus citri is a serious pest of citrus orchards all around the world. In this study, biochemical and resistance inheritance mechanisms of bromopropylate were evaluated on a field population of P. citri. The resistance inheritance of bromopropylate in P. citri has been studied by crossing resistant (As) and susceptible (So) populations, derived from collected populations in northern cities of Iran. The resistance ratios were estimated at 11.45, 3.45, 3.49, 3.72, and 6.16- folds in As, F1, F´1, F2, and F´2 in comparison to the So, respectively. The results of the response showed that resistance was inherited as a bromopropylate incomplete recessive inheritance (D = −0.01 and −0.02) that was detected as polygenic autosomal. The DEM, TPP, and PBO synergism assays in the As population reached 2.27, 2.24, and 1.66- folds, respectively. The in vivo enzyme assays suggested minor involvement of cytochrome P450 monooxygenases, esterases, and GSTs in the P. citri population resistance to bromopropylate, with the highest ratio related to esterases.

Insights into the ALS inhibitor resistance in Amaranthus retroflexus: A matter of concern for CONVISO® SMART sugar beet cultivation

AbstractAmaranthus retroflexus L. has recently become a problematic weed in CONVISO® SMART sugar beet fields in the Czech Republic, largely due to the rapid adoption of this system and widespread application of ALS inhibiting herbicides in almost all crops, including other systems using ALS herbicide tolerant varieties. In this study, two A. retroflexus biotypes, suspected to be resistant to ALS inhibitors, were collected from CONVISO® SMART sugar beet field. Studies were conducted to confirm their resistance status and investigate the underlying mechanisms. The findings reveal that these biotypes have developed resistance to foramsulfuron + thiencarbazone‐methyl, with cross‐resistance to imazamox and tribenuron‐methyl. The dose–response studies revealed that the resistant (R) biotypes AMARE1 and AMARE2 are 351 and 12 times less sensitive to foramsulfuron + thiencarbazone‐methyl than the susceptible (S) biotype. Upon pretreatment with malathion, the resistance index decreased 2X in AMARE1, indicating the potential involvement of cytochrome P450 in the resistance mechanism. However, no involvement of GSTs was detected following NBD‐Cl pretreatment. Furthermore, a higher ALS enzyme activity was observed in both R biotypes compared to the S biotype. Finally, molecular analysis revealed that a single nucleotide substitution had occurred in the ALS gene, conferring a tryptophan (Trp) to Leucine (Leu) substitution at position 574 of the ALS enzyme involved as a probable target‐site resistance mechanism in both R biotypes. In summary, our study identified point mutation and enhanced metabolism by cytochrome P450s are likely mechanisms of resistance to foramsulfuron + thiencarbazone‐methyl in the AMARE1 biotype, while point mutation was found the sole responsible mechanism of resistance involved in AMARE2. The current study reports the first instance of resistance in A. retroflexus against foramsulfuron + thiencarbazone‐methyl. This evolution highlights the dynamic response of weed populations to herbicide pressure and underscores the need for alternative weed control strategies in sugar beet including rotating ALS‐tolerant crops with the traditional (non‐tolerant) crops, to limit ALS inhibitor use and resistance evolution.

Biocatalytic oxyfunctionalization of unsaturated fatty acids to oxygenated chemicals via hydroxy fatty acids.

The selective oxyfunctionalization of unsaturated fatty acids is difficult in chemical reactions, whereas regio- and stereoselective oxyfunctionalization is often performed in biocatalytic synthesis. Fatty acid oxygenases, including hydratases, lipoxygenases, dioxygenases, diol synthases, cytochrome P450 monooxygenases, peroxygenases, and 12-hydroxylases, are used to convert C16 and C18 unsaturated fatty acids to diverse regio- and stereoselective mono-, di-, and trihydroxy fatty acids via selective oxyfunctionalization. The formed hydroxy fatty acids or hydroperoxy fatty acids are metabolized to industrially important oxygenated chemicals such as lactones, green leaf volatiles, and bioplastic monomers, including ω-hydroxy fatty acids, α,ω-dicarboxylic acids, and fatty alcohols, by biocatalysts. For increased oxyfunctionalization of unsaturated fatty acids, enzyme engineering, functional and balanced expression in recombinant cells, selection of suitable catalyst types, and reaction engineering have been suggested. This review describes biocatalysts involved in the oxyfunctionalization of unsaturated fatty acids and the production of hydroxy fatty acids and oxygenated chemicals.

Part 1: Evaluation of Pediatric Cannabis–Drug Interaction Reports

ABSTRACTData addressing safety concerns related to potential drug interactions between cannabis‐derived products and pharmaceutical medications in the pediatric population are lacking. In this study, we retrieved case reports through a published literature search using PubMed and spontaneous reporting data using the Food and Drug Administration's Adverse Event Reporting System (FAERS) to identify potential cannabis– and cannabinoid–drug interactions in individuals younger than 18 years old. To evaluate the published case reports, we used the Drug Interaction Probability Scale (DIPS), a 10‐item questionnaire designed to discern the causal relationship between a potential drug interaction and adverse drug reactions (ADRs). FAERS reports were deduplicated and analyzed to gather information regarding patient demographics, associated drugs, nature of the ADRs, outcomes, professions of the reporters, and reporting timelines. Seven published case reports and 9142 FAERS ADRs reports were included in the final analysis. Based on the findings, caution is warranted when cannabis or cannabinoids are used in combination with prescribed medications, including methadone, everolimus, fluoxetine, and paroxetine. Cannabinoids may inhibit drug‐metabolizing enzymes, including several cytochrome P450s, leading to increased drug exposure and potentially, an increased risk for ADRs.

Non-Canonical Cytochrome P450 Enzymes in Nature.

Cytochrome P450s (CYPs) are a superfamily of thiolate-ligated heme metalloenzymes principally responsible for the hydroxylation of unactivated C-H bonds. The lower-axial cysteine is an obligatory and universally conserved residue for the CYP enzyme class. Herein, we challenge this paradigm by systematically identifying non-canonical CYPs (ncCYPs) that do not harbor a cysteine ligand. Our bioinformatic search reveals 20 distinct ncCYP families with diverse ligands encoded in microbial genomes. We characterize a native serine-ligated CYP with a high-spin ferric resting state. Its crystal structure clearly shows a typical CYP fold and a serine alkoxide as a lower axial heme ligand. In addition, we report the discovery and characterization of the first native selenocysteine-ligated CYP in nature. Our findings radically expand the CYP metalloenzyme family.

Characterisation of the Cytochrome P450 Monooxygenase CYP116B46 from Tepidiphilus thermophilus as a Homogentisic Acid Generating Enzyme and its Conversion to a Peroxygenase.

The heme enzymes of the cytochrome P450 superfamily (CYPs) catalyse the selective hydroxylation of unactivated C-H bonds in organic molecules. There is great interest in applying these enzymes as biocatalysts with a focus on self-sufficient CYP 'fusion' enzymes, comprising a single polypeptide chain with the electron transfer components joined to the heme domain. Here we elucidate the function of the self-sufficient CYP116B46 fusion enzyme, from the thermophilic bacterium Tepidiphilus thermophilus. We demonstrate that it efficiently hydroxylates aromatic organic acids, exemplified by oxidation of 2-hydroxyphenylacetic acid to homogentisic acid (2,5-dihydroxyphenylacetic acid), an important metabolite in bacterial catabolism. In line with the thermophilic nature of the source bacterium, activity increased at higher temperatures, (50 °C), with a catalytic preference for NADPH over NADH. While self-sufficient fusion enzymes simplify biocatalysis; engineered peroxygenase activity is also a key advance in the application of these enzymes as biocatalysts as it eliminates the need for electron transfer partner proteins and nicotinamide cofactors. We demonstrate that a T278E mutation in the heme domain of CYP116B46, confers peroxygenase activity. This engineered peroxygenase enzyme is stable to elevated temperatures and catalytic concentrations of hydrogen peroxide, with an observed optimal activity resulting in a total turnover number of ~650.

The effect of synergists on the inhibition of detoxification enzyme activities and acaricide sensitivity in Rhizoglyphus robini.

The saffron bulb mite, Rhizoglyphus robini Claparede (Acari: Acaridae), is the most important pest of the saffron crop in Iran. This pest attacks and feeds on saffron corms. For this reason, the corms are treated with acaricides before planting. The high activity of detoxification enzymes in arthropods may reduce their pesticide sensitivity. Diethyl maleate (DEM) is an inhibitor of glutathione S-transferases (GSTs), piperonyl butoxide (PBO) is an inhibitor of cytochrome P450 monooxygenases, and triphenyl phosphate (TPP) is an inhibitor of esterase activity. A filter paper method was used to determine the efficiency of these synergists in inhibiting the activity of detoxifying enzymes of R. robini. Adult mites were treated with these three synergists for 6, 12, 24, and 48 h, respectively. The activity of each detoxifying enzyme was measured and compared to the control treatment, and the inhibition percentage was calculated each time. The results showed that DEM reduced GST activity by 59.9% after 48 h, PBO inhibited cytochrome P450 activity by 30%, and TPP suppressed esterase activity by 38.5%. The most statistically significant inhibition occurred 24 h after pretreatment with each synergist. Bioassays with 24 h pretreatment showed that the sensitivity of R. robini to propargite increased by 1.6 times with PBO, 1.7 times with TPP, and 2.5 times with DEM. In conclusion, synergists and efficient inhibition of detoxifying enzymes can play a significant role in increasing the sensitivity of agricultural pests to pesticides and can be considered in managing pesticide resistance.

Genomic Analysis Reveals Novel Genes and Adaptive Mechanisms for Artificial Diet Utilization in the Silkworm Strain Guican No.5.

The transition from traditional mulberry leaf feeding to artificial diet cultivation represents a major advancement in modern sericulture, yet the genetic mechanisms driving this adaptation remain largely unexplored. This study investigates the genomic basis of artificial diet adaptation in the silkworm strain Guican No.5 through whole-genome resequencing and transcriptome analysis. We identified 8,935,179 single-nucleotide polymorphisms (SNPs) across all chromosomes, accounting for 2.01% of the genome, with particularly high densities observed in chromosomes 23, 26, and 28. Our analysis also revealed 879 novel transcripts, many of which are involved in digestion, detoxification, and stress response pathways. Key novel genes, including three carboxylesterases, two cytochrome P450s, one heat shock protein, and one copper/zinc superoxide dismutase, exhibited varying degrees of sequence similarity to known proteins, suggesting modifications to existing genetic frameworks. Notably, one novel P450 gene displayed only 74.07% sequence identity with its closest homolog, indicating the emergence of a new protein sequence. Additionally, several key genes showed high similarity to wild silkworm (Bombyx mandarina) proteins, underscoring their evolutionary origins. These findings provide valuable insights into the molecular mechanisms underpinning artificial diet adaptation in silkworms and offer genomic resources to enhance artificial diet formulations and breeding programs in sericulture.

Functional Expression and Construction of a Self-Sufficient Cytochrome P450 Chimera for Efficient Steroidal C14α Hydroxylation in Escherichia coli.

C14-functionalized steroids enabled diverse biological activities in anti-gonadotropin and anticancer therapy. However, access to C14-functionalized steroids was impeded by the deficiency of chemical synthetic methods. Recently, several membrane-bound fungal cytochrome P450s (CYPs) have been identified with steroid C14α-hydroxylation activity. However, the lack of efficient heterologous overexpression strategy hampered their further characterization and molecular engineering. In the present study, sequences of fungi-derived CYP genes encoding putative 14α-hydroxylase were selected and bioinformatically analyzed. Substitution of the N-terminal hydrophobic helix by a soluble maltose binding protein tag significantly enhanced the soluble expression level in Escherichia coli. A novel CYP originated from Bipolaris oryzae was discovered with high steroidal C14α-hydroxylation activity when coupled with the redox partner CPRlun. A catalytically self-sufficient chimeric CYP-CPR was built by intramolecular fusion, and the electronic transfer rate was improved. A coenzyme NADPH regeneration system was finally constructed by the co-expression of glucose dehydrogenase. The developed soluble multi-enzyme cascade biotransformation system supported the selective C14α-hydroxylation toward progesterone with a final titer of 34.54 mg/L, the highest level achieved in E. coli-based heterologous expression system. This study provides insightful ideas on the functional expression of fungi-derived CYPs and promises an efficient C14α-hydroxylation system for steroidal drugs through protein engineering.

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.

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.