JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2025
·
IF 6.2
Ziyi Chen,Chunyan Yin,Falong Wang,Xinlin Sun,Wishwajith Kandegama,Zhenyu Wang,Guangfu Yang
The green peach aphid (Myzus persicae, Hemiptera: Aphididae) is a globally important pest. Isocycloseram (a novel isooxazoline) and fluralaner (a benzamide/sulfonamide) are novel insecticides. A Hainan-resistant population (HNR), exhibited 459- and 316-fold resistance to isocycloseram and fluralaner, respectively, compared with a Yunnan susceptible population (YNS). Synergist bioassays and enzyme activity analyses indicated that enhanced cytochrome P450 monooxygenase activity contributed to resistance. Transcriptomic and qRT-PCR analyses revealed that CYP6CY3 was upregulated 16.4- and 10.8-fold in HNR compared with YNS. RNA interference of CYP6CY3 significantly restored susceptibility to both insecticides. In vitro recombinant enzyme assays confirmed CYP6CY3-mediated metabolism of isocycloseram and fluralaner, and gene copy number analysis demonstrated a 7.09-fold amplification. These results reveal CYP6CY3-mediated cross-resistance to isocycloseram and fluralaner, representing the first mechanistic evidence of isoxazoline resistance in M. persicae.
研究领域:
昆虫学
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农药科学
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分子生物学
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昆虫生物化学
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毒理学
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DOI
PubMed
高影响因子
BIOCHEMICAL GENETICS
·
2020
·Vol. 58
·期 4
·551-565
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IF 2.027
Xie Shaolin,Zhou Aiguo,Feng Yongyong,Zhang Yue,Li Junyi,Sun Zhuolin,Fan Lanfen,Zou Jixing
Cytochrome P4501A (CYP1A) has been used as a specific biomarker for monitoring water contamination such as PAHs, PCBs and dioxins. In the present study, the cyp1a gene of Gambusia affinis was cloned and sequenced and their expressions under PAHs exposure were characterized. The newly identified cyp1a encodes a protein with 521 amino acids that shared 96–80% identity with other Cyprinodontiformes fishes. RT-PCR analysis revealed that the basal mRNA level of cyp1a was highly expressed in liver and intestine. The expression level of cyp1a was significantly induced by exposure to 100 μg/L 3, 4-Benzopyrene (BaP) for 5 days in the muscle, testis, brain, liver and intestine of adult male fish. Except in the testis, the induced mRNA level of cyp1a ultimately decreased after prolonging the exposure time to 25 days. As for testis, the induced mRNA level of cyp1a was maintained at a high level during the entire exposure time under 100 μg/L BaP exposure. Furthermore, the expression of cyp1a increased with exposure time under a relatively low exposure concentrations 1 μg/L. Regarding the effects of other PAHs, D(a,h)A, BbF, and BaA showed a statistically significant effect of induction on mRNA level of cyp1a in the muscle, testis, brain, liver and intestine.
研究领域:
环境科学
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分子生物学
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DOI
PubMed
高影响因子
Metabolites
·
2019
·Vol. 9
·期 4
·77
·
IF 3.303
Xiao Pu,Yiqiao Gao,Ruiting Li,Wei Li,Yuan Tian,Zunjian Zhang,Fengguo Xu
Cytochrome P450 1A2 (CYP1A2) is one of the major CYP450 enzymes (CYPs) in the liver, and participates in the biotransformation of various xenobiotics and endogenous signaling molecules. The expression and activity of CYP1A2 show large individual differences, due to genetic and environmental factors. In order to discover non-invasive serum biomarkers associated with hepatic CYP1A2, mass spectrometry-based, untargeted metabolomics were first conducted, in order to dissect the metabolic differences in the serum and liver between control rats and β-naphthoflavone (an inducer of CYP1A2)-treated rats. Real-time reverse transcription polymerase chain reaction and pharmacokinetic analysis of phenacetin and paracetamol were performed, in order to determine the changes of mRNA levels and activity of CYP1A2 in these two groups, respectively. Branched-chain amino acids phenylalanine and tyrosine were ultimately focalized, as they were detected in both the serum and liver with the same trends. These findings were further confirmed by absolute quantification via a liquid chromatography–tandem mass spectrometry (LC-MS/MS)-based targeted metabolomics approach. Furthermore, the ratio of phenylalanine to tyrosine concentration was also found to be highly correlated with CYP1A2 activity and gene expression. This study demonstrates that metabolomics can be a potentially useful tool for biomarker discovery associated with CYPs. Our findings contribute to explaining interindividual variations in CYP1A2-mediated drug metabolism.Keywords:CYP1A2;endogenous biomarkers;metabolomics;branched-chain amino acids;phenylalanine;tyrosine
研究领域:
药理学
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生物化学
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DOI
PubMed
高影响因子
Microbial Cell Factories
·
2026
·
IF 4.9
Ma Jinglin,Zhuang Yan,Guan Huiting,Zhang Zhenjun,Zhu Ning,Han Changze,Wang Yonggang,Chen Jixiang
Cytochrome P450 monooxygenases (CYPs) constitute a versatile family of biocatalysts capable of regio- and stereoselective hydroxylation, offering significant potential for pharmaceutical synthesis and the environmental remediation of endocrine-disrupting steroids. Although their synthetic utility is widely recognized, the mechanistic basis of their activity, particularly in bacteria, remains insufficiently explored. Building on prior discovery of steroid-degrading activity in Rhodococcus erythropolis KB1, in this study, we functionally characterized three putative P450 enzymes (CYP125ARh1, CYP125ARh2, and CYP153ARh3) identified from its genome. Substrate-binding assays demonstrated strong affinity toward progesterone and testosterone, but negligible affinity for 11β-hydroxysteroids, including cortisol and corticosterone. In vitro conversion assays revealed that all three enzymes specifically catalyze 16α-hydroxylation of progesterone and testosterone, although with varying catalytic efficiencies. Among them, CYP125ARh2 was identified as the most efficient enzyme, achieving near-quantitative conversion of progesterone (88%) and testosterone (62%). Kinetic analysis further underscored the superior catalytic properties of CYP125ARh2, which displayed lower Michaelis constants (K m = 16.41 ± 1.97 µM for progesterone and 24.50 ± 2.60 µM for testosterone) and higher turnover rates (V max = 6.05 ± 0.16 µM/min and 5.09 ± 0.13 µM/min, respectively). The superior catalytic efficiency of CYP125ARh2 originates from its unique active site architecture, which integrates a pre‑organized, synergistic residue network with pronounced induced‑fit plasticity. This structural synergy enables precise substrate positioning and optimal reaction geometry, thereby rationalizing its high 16α‑hydroxylation activity toward both progesterone and testosterone. These findings highlight CYP125ARh2 as a promising template for engineering high-performance biocatalysts. Overall, this work provides foundational insights into the role of the P450 enzymes in steroid chemistry and the biodegradation of steroidal pollutants.
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PubMed
高影响因子
Communications Biology
·
2019
·Vol. 2
·期 1
·1-11
·
IF 0
Lin Yanke,Wang Shuai,Zhou Ziyue,Guo Lianxia,Yu Fangjun,Wu Baojian
Metabolism is a major defense mechanism of the body against xenobiotic threats. Here we unravel a critical role of Bmal1 for circadian clock-controlled Cyp3a11 expression and xenobiotic metabolism. Bmal1 deficiency decreases the mRNA, protein and microsomal activity of Cyp3a11, and blunts their circadian rhythms in mice. A screen for Cyp3a11 regulators identifies two circadian genes Dbp and Hnf4α as potential regulatory mediators. Cell-based experiments confirm that Dbp and Hnf4α activate Cyp3a11 transcription by their binding to a D-box and a DR1 element in the Cyp3a11 promoter, respectively. Bmal1 binds to the P1 distal promoter to regulate Hnf4α transcriptionally. Cellular regulation of Cyp3a11 by Bmal1 is Dbp- and Hnf4α-dependent. Bmal1 deficiency sensitizes mice to toxicities of drugs such as aconitine and triptolide (and blunts circadian toxicity rhythmicities) due to elevated drug exposure. In summary, Bmal1 connects circadian clock and Cyp3a11 metabolism, thereby impacting drug detoxification as a function of daily time.
研究领域:
药理学
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分子生物学
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时间生物学
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DOI
PubMed
DRUG METABOLISM AND DISPOSITION
·
2018
·Vol. 46
·期 11
·1538-1545
·
IF 3.64
Jiangming Deng,Lianxia Guo,Baojian Wu
Human CYP2A6 (Cyp2a5 in mice) plays an important role in metabolism and detoxification of various drugs and chemicals. Here, we investigated a potential role of peroxisome proliferator-activated receptor γ (Ppar-γ) in circadian regulation of the Cyp2a5 enzyme. We first showed that Cyp2a5 mRNA and protein in mouse liver displayed robust circadian oscillations. Consistent with a circadian protein pattern, Cyp2a5-mediated 7-hydroxylation of coumarin was circadian time–dependent. Formation of 7-hydroxycoumarin was more extensive at a dosing time of Zeitgeber time 2 (ZT2) than that at ZT14. Interestingly, the nuclear receptor Ppar-γ was also a circadian gene. Circadian Ppar-γ protein level was strongly correlated with the Cyp2a5 mRNA level (r = 0.989). Furthermore, Ppar-γ activation (by a selective agonist, rosiglitazone) upregulated Cyp2a5 expression in Hepa-1c1c7 cells, whereas Ppar-γ knockdown downregulated Cyp2a5 expression. Also, Ppar-γ knockdown blunted the rhythmicity of Cyp2a5 mRNA in serum-shocked Hepa-1c1c7 cells. In addition, a combination of promoter truncation analysis, mobility shift, and chromatin immunoprecipitation assays revealed that Ppar-γ directly bound to a PPAR response element (i.e., the −1418- to −1396-bp region) within Cyp2a5 promoter and activated the gene transcription. Taken together, Ppar-γ was a transcriptional activator of Cyp2a5, and its rhythmic expression contributed to circadian expression of Cyp2a5.
研究领域:
药理学
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分子生物学
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昼夜节律生物学
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DOI
PubMed
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2019
·
IF 2.87
Kai Tian,Jiang Zhu,Mei Li,Xinghui Qiu
Capsaicin (8-methyl- N -vanillyl-6-nonenamide) is the most abundant capsaicinoids found in hot peppers ( Capsicum annum and Capsicum frutescens ). It has been well documented that capsaicin plays an important role in the defense against the attack of herbivores or pathogens on Capsicum plants. A few insect herbivores such as Helicoverpa armigera and Helicoverpa assulta have been recorded to be capable of feeding on hot pepper fruits, suggesting that these insects evolve mechanisms against the toxicity of capsaicin. Although cytochrome P450-meidated detoxification is considered to be an important mechanism by which cotton bollworms cope with capsaicin, experimental evidence is lacking. In this study, we compared the capacity of four H. armigera P450s (CYP6B6, CYP9A12, CYP9A14 and CYP9A17) in capsaicin metabolism, and the capsaicin metabolites were screened and tentatively identified by liquid chromatography–tandem mass spectrometry (LC–MS/MS). HPLC analyses showed that depletion rates of capsaicin were 21.9 ± 0.1, 11.9 ± 1.5, 16.3 ± 1.4 and 14.8 ± 0.2 min −1 for CYP6B6, CYP9A12, CYP9A14 and CYP9A17 respectively. The transformation of capsaicin was inhibited by the P450 inhibitor piperonyl butoxide . A total of seven products were detected, and hydroxylation (aromatic and aliphatic) and dehydrogenation were found to be two main pathways in capsaicin metabolism. In addition, capsaicin metabolism was enzyme selective: M1 (ω-hydroxylated N-macrocyclic metabolite) and M3 (ω-hydroxylated metabolite) were uniquely detected in the CYP6B6 catalytic reaction, while M4 (ω-n hydroxylated capsaicin), M5 (diene of capsaicin) and M6 (doubly oxidized metabolite of dehydrogenated capsaicin) were only detectable in CYP9A metabolisms. A capsaicin dimer (5, 5′-dicapsaicin) was found to be the major metabolite of CYP9A reactions, but the minor product produced by CYP6B6. An overall more similar behavior in capsaicin metabolism was observed among CYP9As than between CYP6B6 and CYP9As. Our data demonstrate that CYP6B6 and CYP9As have a potent capability to transform capsaicin, and individual P450 produce unique metabolite profile. These findings help us to understand the molecular basis of capsaicin adaptation in H. armigera .
研究领域:
昆虫学
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植物化学
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分子生物学
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DOI
PubMed
高影响因子
RSC Advances
·
2020
·Vol. 10
·期 11
·6491-6496
·
IF 3.119
Rui-Feng Mei,Ya-Xian Shi,Wei-He Duan,Hao Ding,Xiao-Ran Zhang,Le Cai,Zhong-Tao Ding
α-Terpineol (1), the main volatile constituent in some traditional Chinese medicines, has been reported to be metabolized to 4R-oleuropeic acid by the larvae of common cutworms. The present study verified that α-terpineol could be converted to 4R-oleuropeic acid (2) and (1S,2R,4R)-p-menthane-1,2,8-triol (3) by Alternaria alternata fermentation. Using shortened fermentation times, 7-hydroxy-α-terpineol (2a) was identified as an oxidative intermediate, which was consistent with the hypothesis put forward by previous studies. Cytochrome P450 enzymes were also confirmed to catalyze this biotransformation. This is the first study on the biotransformation of α-terpineol by microbial fermentation.
研究领域:
微生物学
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药理学
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天然产物化学
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DOI
PubMed
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
·
2014
·
IF 2.939
Han-Ming Cui,Qiu-Yan Zhang,Jia-Long Wang,Jian-Long Chen,Yu-Ling Zhang,Xiao-Lin Tong
Ethnopharmacological relevance Berberine (BER) and BER-original herbal medicines have a variety of pharmacological functions and have been widely used in clinical. However, its effect of enzyme induction on cytochrome P450 (CYP) in human hepatocytes is unknown. Material and method Metabolism of berberine and its effect on main metabolic enzymes in HepG2 cell in vitro was investigated. Cocktail probe drugs, mRNA expression and protein expression were used to evaluate the metabolism potency. Meanwhile, an UPLC–MS/MS method was validated for the analysis of BER and four probe drugs in HepG2 cell. Result BER significantly increased the metabolism of midazolam, phenacetin and tolbutamide by inducing the CYP1A2 and 3A4 enzyme in a dose-dependent manner, the mRNA and protein expression of CYP1A2 and 3A4 were increased by berberine at 1000 ng·mL −1 . The activity of CYP1A2 and 3A4 could be induced by BER more than 500 ng·mL −1 in HepG2 cell, which was confirmed by the increase of its mRNA and protein expression. Conclusion BER increases the metabolism of cocktail drugs such as midazolam, phenacetin and tolbutamide by increasing the mRNA and protein expression of CYP1A2 and 3A4.
研究领域:
药理学
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生物化学
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临床医学
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DOI
PubMed
高影响因子
Insects
·
2021
·Vol. 12
·期 7
·651
·
IF 2.769
Xiaorong Xing,Mengwen Yan,Huilin Pang,Fu’an Wu,Jun Wang,Sheng Sheng
Simple SummaryP450s are involved in the detoxification process in insects; however, less attention has been paid for their roles in parasitoid wasps. Here, we identified 28 P450s from the previous constructed transcriptome dataset ofMeteorus pulchricornisand validated their expression patterns under insecticides exposure. Furthermore, we silencedCYP369B3in adult wasps and demonstrated that the knocking downCYP369B3significantly increased of the mortality of adult wasps when they were exposed to each chemical insecticide, phoxim, cypermethrin, and chlorfenapyr, respectively. The present study provides a theoretical base for functional research on P450 genes in parasitoid wasps.AbstractWith the widespread application of insecticides, parasitoid wasps may also be under risk when exposed to insecticides directly at their free-living stages. The endoparasitoid waspMeteorus pulchricornisis the predominant natural enemy of many lepidopteran pests, such asSpodoptera lituraandHelicoverpa armigera. The cytochrome P450 monooxygenases constitute a ubiquitous and complex superfamily of hydrophobic, haem-containing enzymes. P450s are involved in the detoxification of many xenobiotics. However, their exact roles in the tolerance mechanism in parasitoids toward insecticides has received less attention. Here, 28 P450 genes inM. pulchricorniswere identified from a previously constructed transcriptome dataset. These P450 genes belonged to CYP2, -3, and -4, and mitochondrial clans. Subsequently, eight candidateMpulCYPswere selected from four CYP clans to validate their expression patterns under phoxim, cypermethrin, and chlorfenapyr exposure by qRT-PCR. The results showed that all three insecticides had significant effects on the expression ofMpulCYPs. To further study the function of P450s,CYP369B3was silenced, and its expression levels ofCYP369B3were significantly decreased. Survival analysis indicated that after dsRNA injection, the mortality rate of wasps was significantly increased whenM. pulchricornisfemales were exposed to insecticides compared to control groups. Our findings provide a theoretical base for elucidating the mechanism of insecticide tolerance and promote functional research on P450 genes in parasitoid wasps.Keywords:Meteorus pulchricornis;cytochrome P450s;detoxification;insecticide tolerance
研究领域:
昆虫学
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分子生物学
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PubMed
高影响因子
ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY
·
2024
·Vol. 117
·期 3
·e70004
·
IF 1.5
Kai Tian,Jiang Zhu,Xinghui Qiu
Furanocoumarins are a class of chemical compounds with phototoxic properties. For herbivores, efficient detoxification of such defense compounds is the prerequisite to feed successfully on furanocoumarin-containing plants. The cotton bollworm Helicoverpa armigera is a very important polyphagous pest in agriculture, but how it copes with toxic furanocoumarins in some of its host plants is not well understood. Given that cytochrome P450s are well known for their capacity in xenobiotic metabolism, this study attempted to explore the potential roles of cytochrome P450s in furanocoumarin transformation in this pest. Our data showed that two linear structures (psoralen and xanthotoxin) could be metabolized by three recombinant CYP9A enzymes, but no detectable depletion was observed for the linear one with the 8-dimethylallyloxy substituent on the coumarin moiety (imperatorin) and the angular furanocoumarin (angelicin). Initial epoxidation of the double bond connecting C2' and C3' of the furano ring following by cleavage of the epoxidated furan ring, leading to the formation of more soluble, less reactive and nonphotosensitizing metabolites, was identified as a common mechanism of linear furanocoumarin metabolism using a quadrupole/time-of-flight (Q-TOF) mass spectrometry interfaced with a high performance liquid chromatography (HPLC) system. Our data demonstrated that multiple P450s were involved in the detoxification of linear furanocoumarins in the cotton bollworm. These findings contribute to a better understanding of the biochemical basis of adaptation to plant defense chemicals in this economically important pest. Summary CYP9A12, CYP9A14, and CYP9A17 are capable of metabolizing psoralen and xanthotoxin. Epoxidation of the C2'−C3' double bond of the furano ring is the common pathway. The three CYP9A enzymes exhibit no detectable activity against angelicin and imperatorin.
研究领域:
昆虫学
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生化与分子生物学
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产品关联
DOI
PubMed
高影响因子
PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2024
·
IF 4.2
Tao Li,Lisha Yuan,Dun Jiang,Shanchun Yan
Strong multi-host adaptability significantly contributes to the rapid dissemination of Hyphantria cunea. The present study explores the involvement of cytochrome P450 monooxygenase (P450) in the multi-host adaptation of H. cunea and aims to develop RNA pesticides targeting essential P450 genes to disrupt this adaptability. The results showed that inhibiting P450 activity notably reduced larval weight and food-intake across seven plants groups. The P450 gene HcCYP6AE178 was highly upregulated in H. cunea larvae from medium- and low-preference host plant groups. Silencing HcCYP6AE178 significantly decreased H. cunea larval body weight, increased larval mortality, inhibited energy metabolism genes expression and interfered with growth regulatory genes expression. Overexpression of HcCYP6AE178 enhanced the tolerance of Drosophila and Sf9 cells to the plant defensive substances cytisine and coumarin. The RNA pesticide CS-ds HcCYP6AE178 constructed using chitosan (CS) exhibited remarkable stability. Treatment with CS-ds HcCYP6AE178 effectively reduced H. cunea larval body weight, heightened larval mortality, and disrupted growth regulatory genes expression in low-preference host plant groups. Combined treatment of CS-ds HcCYP6AE178 and coumarin significantly elevated H. cunea larval mortality compared to coumarin alone, accompanied by the inhibition of growth regulatory genes expression and an abnormal increase in energy metabolism genes expression. Taken together, HcCYP6AE178 is essential for the adaptation of H. cunea to multiple host plants, and RNA pesticides targeting HcCYP6AE178 can effectively impair the performance of H. cunea in different host plants.
研究领域:
昆虫学
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分子生物学
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植物保护
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DOI
高影响因子
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