PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2019
·
IF 2.87
Bojun Yan,Yuhua Zhang,Jun Li,Jiapeng Fang,Tingting Liu,Liyao Dong
Cytochrome P450s (P450s) confer resistance against herbicides, and this is increasingly becoming a concern for weed control. As a widespread Gramineae weed in paddy fields , Echinocloa glabrescens has become resistant to the acetolactate synthase (ALS)-inhibiting triazolopyrimidine herbicide penoxsulam. In this study, we found that the GR 50 of the resistant population (SHQP-R) decreased substantially from 25.6 to 5.0 and 6.2 g a.i. ha −1 after treatment with the P450 inhibitors piperonyl butoxide (PBO) and malathion , respectively. However, P450 inhibitors almost had no effects on the susceptibility of the sensitive population (JYJD-S) to penoxsulam. To investigate the mechanisms of metabolic resistance, transcriptome sequencing analysis was performed to find candidate genes that may confer resistance to penoxsulam in E. glabrescens . A total of 233 P450 differentially expressed genes (DEGs) were identified by transcriptome sequencing. We found that the metabolic process and metabolic pathways were the most highly enriched in DEGs. Further, twenty-seven candidate P450 DEGs were selected for qPCR validation analyses. After penoxsulam treatment, the relative expression levels were significantly higher in SHQP-R than in JYJD-S. Among these, the relative expression of twenty-three P450 DEGs (eighteen from the CYP72A-71C-74A-96A-734A subfamily; five from CYP81E1-94C1–94B3-714C1-714C2) were upregulated and four P450 DEGs (from CYP724B1-711A1-707A7-97B2) were downregulated. Changes in the expression of these candidate P450 genes in E. glabrescens were in response to penoxsulam, which provides preliminary evidence for the role of P450s in herbicide metabolism in E. glabrescens. However, further functional studies on metabolic resistance to penoxsulam in a resistant E. glabrescens population are required.
研究领域:
植物科学
·
分子生物学
·
杂草科学
·
药理学
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产品关联
DOI
PubMed
高影响因子
PHARMACEUTICAL BIOLOGY
·
2018
·
IF 1.918
Huaqiao Tang,Nana Long,Min Dai,Lin Lin,Jianlong Li,Fenghui Sun,Lijuan Guo
Context: Citral is used as a potential natural treatment for various infectious diseases.Objective: To examine the effect of citral on the mRNA expression and activities of cytochrome P450 (CYP450) enzymes and establish the relationship between citral-induced liver injury and oxidative stress.Materials and methods: ICR mice were randomly divided into citral (20, 200, and 2000 mg/kglow), Tween-80, and control groups (0.9% saline), 10 mice in each group. The citral-treated groups were intragastrically administered citral for 3 d, control groups treated with 0.5% Tween-80 and 0.9% saline in the same way. Liver injury and CYP450 enzymes were analyzed by analyzing the histopathological changes and the changes of related enzymes.Results: Citral treatment (2000 mg/kg) for 3 d increased serum glutamic pyruvic transaminase and glutamic oxaloacetic transaminase levels, as well as glutathione, gydroxyl radicals, malonaldehyde and total superoxide dismutase contents, but decreased the content of total antioxidant capacity. In doses of 20 and 200 mg/kg groups mice, the contents of NO were decreased significantly and other changes were similar to the 2000 mg/kg group mice, but the liver damage was most severe in the 2000 mg/kg group. Citral induced the mRNA expression and activities of CYP450 1A2, 2D22, and 2E1 in the liver of mice at doses of 20 and 200 mg/kg. There were no changes in testing indexes in Tween-80 treated group mice. Due to its toxic effects, the CYP induction effect of citral negatively correlated with its dose. Although the mRNA expression of CYP450 3A11 was induced by citral, its activity was not affected by low and moderate doses of citral. CYP450 3A11 activity was significantly decreased by high-dose citral.Conclusions: Citral is hepatotoxic and induced oxidative stress in higher dose, which has a negative effect on CYP450 enzymes. These data suggest caution needs to be taken in order to avoid citral-drug interactions in human beings.
研究领域:
药理学
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毒理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF TOXICOLOGICAL SCIENCES
·
2014
·Vol. 39
·期 4
·609-614
·
IF 1.378
Xiao Run-mei,Wang Jun-jun,Chen Jing-ya,Sun Li-juan,Chen Yong
Betel-quid use is associated with the risk of liver cirrhosis and hepatocellular carcinoma. The aim of the present work was to evaluate the impact of arecoline on human hepatic cytochrome P450 (CYP) enzymes in vitro and rat hepatic CYP enzymes, as well as the hepatic oxidative stress and liver injury of rats in vivo. The in vitro results indicated that arecoline hydrobromide (AH) has no significant effect on the activities of CYP2B, 2C9, 3A4, 1A2, 2E1 and 2D6 in human liver microsome (HLM). However, oral administration of AH at 4 and 20 mg/kg/d for seven consecutive days significantly increased the activities of rat hepatic CYP2B, 2E1, 2D, 3A, 2C and 1A2. In addition, AH at 100 mg/kg/d significantly increased the levels of ALT, AST and MDA, decreased the levels of SOD, CAT, GSH-Px and GSH, in rat liver. The in vivo induction of AH on rat hepatic CYP isoforms suggested that the high risk of metabolic interaction should be existed when the substrate drugs of the six kinds of CYP isoforms was administered in betel-quid use human. Furthermore, the in vivo results also suggested that AH-induced hepatoxicity should be associated with the induction of AH on rat hepatic CYP2E1 and 2B.
研究领域:
药理学
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毒理学
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肝脏病学
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产品关联
DOI
PubMed
高影响因子
CHEMISTRY-A EUROPEAN JOURNAL
·
2023
·Vol. 30
·期 10
·e202302936
·
IF 4.3
Feiyan Yuan,Jing Ding,Yiyang Sun,Jianhua Liang,Yunzi Luo,Yang Yu
Graphical The carbene transfer reaction was realized in the biosynthetic pathway. Using L-limonene as the precursor, L-carveol was biotransformed by P411-PFA to generate unnatural terpenoid derivatives with trifluoromethyl group. Protein engineering of cytochrome P450s has enabled these biocatalysts to promote a variety of abiotic reactions beyond nature‘s repertoire. Integrating such non-natural transformations with microbial biosynthetic pathways could allow sustainable enzymatic production of modified natural product derivatives. In particular, trifluoromethylation is a highly desirable modification in pharmaceutical research due to the positive effects of the trifluoromethyl group on drug potency, bioavailability, and metabolic stability. This study demonstrates the biosynthesis of non-natural trifluoromethyl-substituted cyclopropane derivatives of natural monoterpene scaffolds using an engineered cytochrome P450 variant, P411-PFA. P411-PFA successfully catalyzed the transfer of a trifluoromethyl carbene from 2-diazo-1,1,1-trifluoroethane to the terminal alkenes of several monoterpenes, including L-carveol, carvone, perilla alcohol, and perillartine, to generate the corresponding trifluoromethylated cyclopropane products. Furthermore, integration of this abiotic cyclopropanation reaction with a reconstructed metabolic pathway for L-carveol production in Escherichia coli enabled one-step biosynthesis of a trifluoromethylated L-carveol derivative from limonene precursor. Overall, amalgamating synthetic enzymatic chemistry with established metabolic pathways represents a promising approach to sustainably produce bioactive natural product analogs.
研究领域:
生物化学
·
合成生物学
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药物化学
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DOI
PubMed
高影响因子
CHEMISTRY & BIODIVERSITY
·
2021
·Vol. 18
·期 4
·e2000910
·
IF 2.408
Bingbing Ma,Qianwen Wang,Bing-Nan Han,Haruo Ikeda,Chunfang Zhang,Lian-Hua Xu
Cytochrome P450 enzymes (P450s) are versatile biocatalysts, which insert a molecular oxygen into inactivated C−H bonds under mild conditions. CYP105D7 from Streptomyces avermitilis has been reported as a bacterial substrate-promiscuous P450 which catalyzes the hydroxylation of 1-deoxypentalenic acid, diclofenac, naringenin, compactin and steroids. In this study, CYP105D7 catalyzes hydroxylation, epoxidation and dehydrogenation of capsaicin, a pharmaceutical agent, revealing its functional diversity. The kinetic parameters of the CYP105D7 oxidation of capsaicin were determined as K m =311.60±87.30 μM and k cat =2.01±0.33 min −1 . In addition, we conducted molecular docking, mutagenesis and substrate binding analysis, indicating that Arg81 plays crucial role in the capsaicin binding and catalysis. To our best knowledge, this study presents the first report to illustrate that capsaicin can be catalyzed by prokaryotic P450s.
研究领域:
生物化学
·
药物代谢
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DOI
PubMed
高影响因子
CHEMBIOCHEM
·
2014
·Vol. 15
·期 16
·2443-2449
·
IF 3.06
Yue-Cai Yin,Hui-Lei Yu,Zheng-Jiao Luan,Ren-Jie Li,Peng-Fei Ouyang,Jing Liu,Jian-He Xu
Graphical Catalytic versatility of P450 La MO : A new redox-self-sufficient P450 of the CYP116B subfamily was discovered by data mining. A substrate spectrum study showed that it was a catalytic versatile monooxygenase, mediating oxidation reactions as diverse as hydroxylation, alkene epoxidation, O -dealkyl- ation, sulfoxidation, and even hydration. A new member of the CYP116B subfamily—P450 La MO —was discovered in Labrenzia aggregata by genomic data mining. It was successfully overexpressed in Escherichia coli , purified, and subsequently characterized spectroscopically, and its catalytic properties were assessed. Substrate profiling of the P450 La MO revealed that it was a versatile catalyst, exhibiting hydroxylation and epoxidation activities as well as O -dealkylation and asymmetric sulfoxidation activities. Diverse compounds, including alkylbenzenes, aromatic bicyclic molecules, and terpenoids, were shown to be hydroxylated by P450 La MO . Such diverse catalytic activities are uncommon for the bacterial P450s, and the P450 La MO -mediated stereoselective hydroxylation of inactivated C H bonds—ubiquitous and relatively unreactive in organic molecules—is particularly unusual. The self-sufficient nature of P450 La MO , coupled with its broad substrate range, highlights it as an ideal template for directed evolution towards various applications.
研究领域:
生物化学
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酶学
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微生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2019
·
IF 3.571
Zi-Guo Wang,Shan-Shan Jiang,David Mota-Sanchez,Wei Wang,Xin-Ru Li,Yu-Lin Gao,Xu-Peng Lu,Xue-Qing Yang
Resistance to pyrethroid and organophosphate insecticides has been a growing problem in the management of cotton bollworm Helicoverpa armigera (Hübner) populations in the Yangtze River and Yellow River valleys of China, but resistance status and mechanisms of H. armigera populations from northeast China are less documented. In this study, a field strain collected from Shenyang in northeast China (SYR) is up to 16-fold more resistant than a susceptible strain (SS) to λ-cyhalothrin, while the resistance level to phoxim remained low (2.6-fold). Synergist tests and enzymatic assays show that increased cytochrome P450 monooxygenase (P450) activity is the main mechanism for λ-cyhalothrin resistance. Eight out of 10 genes from CYP6 and CYP9 subfamilies were significantly overexpressed in the SYR strain, and CYP6AE11 was the mostly overexpressed P450 (59-fold). These results suggest that overexpression of multiple P450 enzymes contributes to λ-cyhalothrin resistance in the SYR strain of H. armigera from northeast China.
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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产品关联
DOI
PubMed
高影响因子
ACS Catalysis
·
2025
·
IF 13.1
Tianjian Sun,Mingyu Liu,Shuai Li,Jiawei Guo,Lei Liu,Lixin Zhang,Li Ma,Shengying Li
Avermectins (AVMs) are a class of 16-membered ring macrolides produced by Streptomyces avermitilis. Renowned for their potent insecticidal and acaricidal properties, AVMs are widely used as environmentally friendly biopesticides. Although the biosynthetic gene aveE encoding a cytochrome P450 monooxygenase was identified 30 years ago, its exact catalytic function and mechanism have remained elusive due to a lack of biochemical characterization. Here, we overcome the long-standing challenge in soluble and functional protein expression of AveE in Escherichia coli and reconstitute the in vitro activity of this P450 enzyme using surrogate redox partner proteins. Time-course studies reveal monohydroxylation at the C8a position of the substrates as the initial step, subsequently leading to tetrahydrofuran (THF) ring formation. Isotopic labeling experiments provide significant insight into the catalytic mechanism of AveE, revealing the cooperation of the C8a,C6-diol pathway and the C6-nucleophilic attack pathway. This study not only presents an effective strategy for heterologous expression of difficult P450 enzymes in E. coli but also elucidates the exact process of THF ring formation during AVM biosynthesis.
研究领域:
微生物学
·
酶学
·
生物化学
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2022
·
IF 5.895
Zhuang Zhang,Zanrong Wen,Kaixin Li,Wei Xu,Ni Liang,Xinyue Yu,Changyou Li,Dong Chu,Lei Guo
Bemisia tabaci is an important agricultural sucking pest, and it develops serious resistance to various insecticides. Although cytochrome P450 was involved in the resistance to cyantraniliprole, limited studies have been conducted on B. tabaci. In the present study, piperonyl butoxide significantly increased the toxicity of cyantraniliprole. P450 activities in two resistant populations were 1.97- and 2.17-fold higher than that in the susceptible population. Among 79 P450 genes, CYP6CX3 expressions in two resistant populations were 3.08- and 3.67-fold higher than that in the susceptible population. When CYP6CX3 was knocked down, the toxicity of cyantraniliprole increased significantly. The LC50 value of cyantraniliprole to the Drosophila melanogaster line overexpressing B. tabaciCYP6CX3 increased 7.34-fold. The content of cyantraniliprole was decreased by 25.74 ± 4.27% after mixing with CYP6CX3 and CPR for 2 h. These results suggested that the overexpression of CYP6CX3 was likely involved in the resistance to cyantraniliprole in B. tabaci.
研究领域:
农业昆虫学
·
分子生物学
·
农药科学
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产品关联
DOI
PubMed
高影响因子
TOXICOLOGY LETTERS
·
2025
·
IF 2.9
Siyu Liu,Guode Zhao,Yingyun Xu,Yang Wang,Zifang Ding,Weiwei Li,Ying Peng,Jiang Zheng
Osthole (OST), a coumarin derivative, is one of the major components of Cnidium monnieri (L.) Cussion. OST was reported to induce apoptosis in hepatocytes. Elevated serum ALT and AST were documented in Sprague-Dawley rats after administration of OST. In the present study, OST was found to be metabolized to a phenol metabolite which was further metabolically oxidized to the corresponding quinone methide intermediate. A glutathione conjugate derived from the reactive metabolite was detected in vitro and in vivo . The structures of the metabolites were verified by chemical analysis. CYP3A4 and CYP1A2 were the major enzymes to catalyze the oxidation reactions. Pre-treatment with 1-aminobenzotriazole or ketoconazole decreased the susceptibility of primary hepatocytes to the cytotoxicity of OST. The findings provided solid evidence that the metabolic activation of OST correlated with the cytotoxicity of OST.
研究领域:
药理学
·
毒理学
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产品关联
DOI
PubMed
高影响因子
PLANTA
·
2021
·Vol. 254
·期 5
·1-10
·
IF 4.116
Wang Jiadian,Su Ping,Gao Linhui,Zhang Yifeng,Wang Jian,Tu Lichan,Zhao Yujun,Lu Yun,Yin Yan,Huang Luqi,Gao Wei
Main conclusion A novel cytochrome P450 from Tripterygium wilfordii , CYP81AM1, specifically catalyses the C-15 hydroxylation of dehydroabietic acid. This is the first CYP450 to be found in plants with this function. Abstract Cytochrome P450 oxygenases (CYPs) play an important role in the post-modification in biosynthesis of plant bioactive terpenoids. Here, we found that CYP81AM1 can catalyze the formation of 15-hydroxydehydroabietic acid by in vitro enzymatic reactions and in vivo yeast feeding assays. This is the first study to show that CYP81 family enzymes are involved in the hydroxylation of abietane diterpenoids. At the same time, we found that CYP81AM1 could not catalyse abietatriene and dehydroabietinol, suggesting that the occurrence of the reaction may be related to the carboxyl group. Through molecular docking and site mutations, it was found that some amino acid sites (F104, K107) near the carboxyl group had an important effect on enzyme activity, also suggesting that the carboxyl group played an important role in the occurrence of the reaction.
研究领域:
植物学
·
生物化学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2018
·Vol. 19
·期 7
·2140
·
IF 3.687
Huaqiao Tang,Min Xu,Fei Shi,Gang Ye,Cheng Lv,Jie Luo,Ling Zhao,Yinglun Li
Although nano-copper is currently used extensively, the adverse effects on liver cytochrome P450 (CYP450) enzymes after oral exposure are not clear. In this study, we determined the effects and mechanisms of action of nano- and micro-copper on the expression and activity of CYP450 enzymes in rat liver. Rats were orally exposed to micro-copper (400 mg/kg), Cu ion (100 mg/kg), or nano-copper (100, 200 and 400 mg/kg) daily for seven consecutive days. Histopathological, inflammatory and oxidative stress were measured in the livers of all rats. The mRNA levels and activity of CYP450 enzymes, as well as the mRNA levels of select nuclear receptors, were determined. Exposure to nano-copper (400 mg/kg) induced significant oxidative stress and inflammation relative to the controls, indicated by increased levels of interleukin (IL)-2, IL-6, interferon (IFN)-γ, macrophage inflammatory protein (MIP-1), total antioxidant capacity (T-AOC), malondialdehyde (MDA), inducible nitric oxide synthase (iNOS) and nitric oxide (NO) after exposure. The levels of mRNA expression of pregnane X receptor (PXR), constitutive androstane receptor (CAR) and aryl hydrocarbon receptor (AHR) were significantly decreased in 400 mg/kg nano-copper treated rats. Nano-copper activated the expression of the NF-kappa B (NF-κB), mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription (STAT)3 signaling pathways. Nano-copper decreased the mRNA expression and activity of CYP 1A2, 2C11, 2D6, 2E1 and 3A4 in a dose-dependent manner. The adverse effects of micro-copper are less severe than those of nano-copper on the CYP450 enzymes of rats after oral exposure. Ingestion of large amounts of nano-copper in animals severely affects the drug metabolism of the liver by inhibiting the expression of various CYP450 enzymes, which increases the risk of drug-drug interactions in animals.Keywords:nano-copper;oral exposure;liver toxicity;CYP450 enzymes;mechanism
研究领域:
毒理学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）