PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2026
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IF 4.8
Xinyao Chen,Xuekai Shi,Yuemin Ma,Yichao Zhang,Haihua Wu
Cytochrome P450 monooxygenases (P450s) play pivotal roles in diverse insect physiology, including growth, development, xenobiotic detoxification, and olfaction. In the present study, we identified and characterized a novel P450, LmCYP3136A1, from Locusta migratoria , which lacks the conserved cysteine residue critical for canonical P450 activity in its heme-binding motif. Phylogenetic analyses indicated that LmCYP3136A1 clusters within the CYP3 clan. LmCYP3136A1 was predominantly expressed in the gastric caecum and midgut, with moderate expression in the antenna, and its transcript levels peaked in the 5th-instar nymphal stage. Its expression was significantly induced 6.22-fold by carbaryl and 9.44-time by chlorpyrifos and was also upregulated 1.51-fold by the aggregation pheromone 4-vinylanisole (4-VA) and 1.41-fold by the plant volatile 2-ethylpyrazine. RNAi-mediated silencing enhanced locust susceptibility to malathion and carbaryl, increasing mortality by 35% and 42%, respectively, but not to deltamethrin and chlorpyrifos. Furthermore, LmCYP3136A1 silencing significantly reduced electroantennogram (EAG) responses to eight volatiles, including 4-VA and decanal, with reductions of up to 83%. Molecular docking revealed ligand binding via hydrogen bonds and π-π stacking. Our findings demonstrate putative dual roles for LmCYP3136A1 in mediating specific insecticide detoxification and regulating olfactory sensitivity, expanding understanding of P450 functional diversity and structural plasticity in insects.
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INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
·
2026
·
IF 4.3
Haolin Li,Xue Kong,Han Yang,Dongyu Yang,Jiguang Wei,Xuesheng Li
Juvenile hormones (JHs) play a crucial role in regulating development and reproduction in insects. In the cowpea aphid ( Aphis craccivora ), JH Ⅲ acid (JH ⅢA) can be converted into JH Ⅲ skipped bisepoxide (JHSB 3 ); however, the enzymes and pathways involved in this conversion remain elusive. In this study, we identified Ac -CYP6A14 as a key JH epoxidase and characterized its role in the JHSB 3 biosynthetic pathway in A. craccivora . In addition, we found that JHSB 3 titers were significantly higher in embryonic tissues than in maternal tissues. Transcriptomic analysis revealed six upregulated genes in the embryo. Temporal expression pattern analysis revealed that only Ac - CYP6A14 showed concordance with JHSB 3 titers. Silencing Ac - CYP6A14 decreased JHSB 3 titers. In vitro catalytic experiments demonstrated that Ac -CYP6A14 catalyzes the conversion of JH Ⅲ into JHSB 3 . Furthermore, we found that the “epoxidation then esterification” sequence is not feasible. Collectively, our findings elucidated the JHSB 3 biosynthetic pathway, in which JH ⅢA is first esterified to form JH Ⅲ, followed by epoxidation to JHSB 3 by Ac -CYP6A14.
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Revista Brasileira de Farmacognosia-Brazilian Journal of Pharmacognosy
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2023
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IF 1.6
Huang Mingjun,Wu Fanchang,Zuo Xue,Liu Jinman,Yu Wenqian,Xie Ruiye,Liu Guoqing,Tan Qiwen,Wang Qi,Liang Yong
Plant-based medicines derived from traditional Chinese medicine have garnered widespread attention for their potential in the prevention and treatment of various diseases, offering superior oral bioavailability compared to the use of single drug formulations. Nonetheless, the precise biological mechanisms and interactions between compounds and enzymes remain inadequately understood. This study delves into the inhibitory interactions of quercetin with cytochrome P450 and P-glycoprotein, leading to a synergistic enhancement of osthole bioavailability in rat plasma. The results indicate that quercetin demonstrates concentration-dependent and uncompetitive inhibition of liver microsomal cytochrome P450 proteins 3A, with an observed IC 50 value of 34.17 μM. Additionally, quercetin notably suppresses the expression of P-glycoprotein in the small intestines of rats, resulting in a significant 52.9% reduction ( p < 0.001) and a corresponding downregulation of its associated mRNA Mdr1b level. The administration of quercetin alongside osthole in rats leads to a marked increase of over 80% ( p < 0.001) in osthole blood concentration, as determined by pharmacokinetic analysis, surpassing the effects of pure osthole usage. This multifunctional attribute of quercetin, along with the observed synergistic changes in pharmacokinetics, holds promise for shedding light on the interplay of drug–drug/enzyme interactions in the context of medicinal plants. Graphical abstract
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Expert Opinion on Drug Metabolism & Toxicology
·
2015
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IF 2.831
Danyi Lu,Dong Dong,Zhong Liu,Yifei Wang,Baojian Wu
Objective: The aim of this article is to elucidate the metabolic pathways of BJ-B11, a heat shock protein 90 inhibitor, in human liver microsomes (HLM) and determine the main enzymes responsible for formation of each metabolite.Methods: Metabolites of BJ-B11 were identified using the ultra performance liquid chromatography- quadrupole time-of-flight/mass spectrometry (UPLC-QTOF/MS) method. Esterase contributing to the hydrolysis of BJ-B11 was identified by chemical inhibition and activity correlation assays. Reaction phenotyping and kinetic studies using expressed cytochrome P450 (CYP) enzymes were performed to determine the contributions of CYP isozymes to BJ-B11 metabolism.Results: BJ-B11 was rapidly hydrolyzed to generate a deacetylated product M1-1. M1-1 was subsequently metabolized to form eight metabolites. Hydrolysis of BJ-B11 was markedly inhibited by vinblastine (a dual inhibitor of arylacetamide deacetylase and carboxylesterase 2). By contrast, digitonin and telmisartan (the specific inhibitors for carboxylesterase 1 and carboxylesterase 2, respectively) did not inhibit BJ-B11 hydrolysis at all. Further, BJ-B11 hydrolysis was significantly correlated with hydrolysis of phenacetin (an activity marker of arylacetamide deacetylase). Moreover, reaction phenotyping revealed that metabolism of M1-1 in HLM was attributable to several CYP enzymes, including CYP1A1, 1B1, 3A4 and 3A5.Conclusion: BJ-B11 was subjected to efficient metabolism in the liver, generating nine metabolites. BJ-B11 metabolism was contributed mainly by arylacetamide deacetylase and multiple CYP enzymes.
研究领域:
药理学
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生物化学
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Systems Microbiology and Biomanufacturing
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2023
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IF 0
Li Xuemei,Zhang Rui,Li Jianjiong,Feng Jinhui,Wu Qiaqing,Zhu Dunming,Ma Yanhe
Cholesterol and phytosterols can be served as the substrates for Actinobacteria to produce the precursors used in the synthesis of steroidal medicines. Cytochrome P450s from CYP125 and CYP142 families initiates the metabolism of cholesterol and/or phytosterols. In this study, we demonstrate the functional redundancy and substrate preference of five cytochrome CYP125s and CYP142s by the unmarked cyp genes deletion strains of Mycobacterium neoaurum NRRL B-3805. CYP125-3 was found to be responsible for the catalytic hydroxylation of both cholesterol and the C24-branched side-chains of phytosterols, while CYP142-2 can only oxidize cholesterol and/or cholest-4-en-3-one. The strain harboring the CYP125-3 produced 6.47 g l − 1 and 5.13 g l − 1 of 4-androstene-3,17-dione (AD) when 15 g l − 1 cholesterol or 17.5 g l − 1 phytosterols used, respectively. While, the strain harboring the CYP142-2 produced 7.10 g l − 1 AD when 15 g l − 1 cholesterol used. The other three cytochrome P450s have a smaller contribution for the degradation of cholesterol and phytosterols. CYP125-3 has the hydroxylation activity on more substrates than that of CYP142-2. This study provided possible guidance to improve the efficiency of AD and other metabolites production from different kinds of sterols by the genetic manipulation.
研究领域:
微生物学
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生物化学
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分子生物学
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生物技术
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PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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2025
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IF 4
Jing Liu,Zhengjie Wang,Shuo Yin,Zhonghao Wang,Meng Wu,Jianhua Qin,Junlong Liu,Chuanwen Wang
Dermanyssus gallinae is a significant haematophagous ectoparasite of laying hens. Despite the worldwide application of beta-cypermethrin for mite control, resistance to this compound has been developed by D. gallinae . P450s, as key detoxification enzymes involved in the metabolism of exogenous and endogenous chemical substances, have not been functionally characterized in D. gallinae . Here, the role of the P450s gene (Deg-CYP-3) of D. gallinae in acaricide resistance was studied. Firstly, P450s activities of the five beta-cypermethrin resistant strains (RR) against p-nitroanisole were significantly higher than that of the susceptible strain (RS), suggesting enhanced enzymatic activities might contribute to mite beta-cypermethrin resistance. Both mRNA expression level and DNA copy number of Deg-CYP-3 were significantly higher in RR than in RS, except for DK RR strain. Moreover, recombinant Deg-CYP-3 (rCYP-3) expressed in Escherichia coli displayed high enzymatic activity against p-nitroanisole, while Qulaton-100, acetone, isoacetone, avermectin, and ivermectin were found to inhibit this activity. Immunohistochemical analysis revealed the presence of Deg-CYP-3 protein in the digestive tract and Malpighian tubules. Finally, silencing of Deg-CYP-3 via RNAi resulted in decreased enzyme activity and increased susceptibility of PG RR strain to beta-cypermethrin, confirming that Deg-CYP-3 is crucial for beta-cypermethrin detoxification. RNAi also resulted in enhanced mortality, as well as reduced oviposition and fecundity in the PG RR strain. Collectively, these findings indicate that resistance of D. gallinae to beta-cypermethrin is associated with elevated P450s protein activity and increased Deg-CYP-3 expression levels. The results provide insights into the metabolic resistance mechanisms of D. gallinae and offer scientific guidance for the management and control of mite.
研究领域:
寄生虫学
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农药毒理学
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分子生物学
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兽医昆虫学
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生物化学
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ACS Synthetic Biology
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2025
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IF 3.9
Baodong Hu,Jingwen Zhou,Jianghua Li,Jian Chen,Guocheng Du,Fang Zhong,Yucheng Zhao,Xinrui Zhao
Marmesin, a plant dihydrofuranocoumarin, is an important intermediate in the synthesis of linear furanocoumarins and exhibits a variety of pharmacological activities. However, due to the lack of efficient prenyltransferases, the incompatibility of redox partners for P450 enzymes, and the insufficient supply of precursor (DMAPP), the microbial synthesis of marmesin remained at an extremely low level. Here, we report the efficient biosynthesis of marmesin in Escherichia coli by screening the robust 6-prenyltransferase PpPT1 and marmesin synthase PpDCΔ2–29 from Peucedanum praeruptorum. Next, the activities of PpPT1 and PpDCΔ2–29 were enhanced using fusion protein tags and redox partner engineering, respectively. In addition, the synthesis of marmesin was further improved by strengthening the methylerythritol phosphate (MEP) pathway to increase the availability of DMAPP and by optimizing the modular pathway in the engineered strain. Finally, the titer of marmesin reached 203.69 mg L–1 in the fed-batch fermentation with a molar conversion rate of umbelliferone of 81.4%, which is the highest titer for marmesin production using engineered microorganisms. The applied strategy and marmesin-producing strain constructed in this study lay the foundation for the green production of valuable complex furanocoumarins.
研究领域:
生物化学
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合成生物学
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药理学
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METABOLIC ENGINEERING
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2025
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IF 6.8
Dan He,Yan Chen,Junfeng Shen,Han Yu,Jay D. Keasling,Xiaozhou Luo
10-hydroxy-2-decenoic acid (10-HDA), a unique unsaturated fatty acid present in royal jelly, has attracted considerable interest due to its potential medical applications. However, its low concentration in royal jelly and complex conformational structure present challenges for large-scale production. In this study, we designed and constructed a de novo biosynthetic pathway for 10-HDA in Escherichia coli . Initially, we introduced the heterologous thioesterase UaFatB1 to hydrolyze trans-2-decenoyl ACP to produce trans-2-decenoic acid, a key precursor for 10-HDA. Subsequently, we employed the bacterial cytochrome P450 enzyme CYP153AMaq to catalyze the terminal hydroxylation of trans-2-decenoic acid. Furthermore, through redox partner engineering and directed evolution, we identified the optimal combination for 10-HDA production: CYP153AMaq Q129R/V141L mutant with redox partner FdR0978/Fdx0338. Finally, we optimized the fermentation conditions and achieved a 10-HDA titer of 18.8 mg/L using glucose as primary carbon source. Our work establishes a platform for producing α , β -unsaturated fatty acids and their derivatives, facilitating further studies on these compounds.
研究领域:
生物化学
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合成生物学
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微生物学
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ENVIRONMENTAL POLLUTION
·
2024
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IF 7.6
Guangcai Ma,Kan Ma,Jing Zhang,Xianglong Zhao,Qiuyi Wang,Yewen Chen,Jiayu Lu,Xiaoxuan Wei,Xueyu Wang,Haiying Yu
The escalating focus on the environmental occurrence and toxicology of emerging pollutants underscores the imperative need for a profound exploration of their metabolic transformations mediated by human CYP450 enzymes. Such investigations have the potential to unravel the intricate metabolite profiles, substantially altering the toxicological outcomes. In this study, we integrated the computational simulations with in vitro metabolism experiments to investigate the metabolic activity and mechanism of an emerging pollutant, 1,3,5-tris(2,3-dibromopropyl)-1,3,5-triazinane-2,4,6-trione (TDBP-TAZTO), catalyzed by human CYP450s. The results highlight the important contributions of CYP2E1, 3A4 and 2C9 to the biotransformation of TDBP-TAZTO, leading to the identification of four distinct metabolites. The effective binding conformations governing biotransformation reactions of TDBP-TAZTO within active CYP450s are unveiled. Structural instability of primary hydroxyTDBP-TAZTO products suggests three potential outcomes: (1) generation of an alcohol metabolite through successive debromination and reduction reactions, (2) formation of a dihydroxylated metabolite through secondary hydroxylation by CYP450, and (3) production of an N-dealkylated metabolite via decomposition and isomerization reactions in the aqueous environment. The formation of a desaturated debrominated metabolite may arise from H-abstraction and barrier-free Br release during the primary oxidation, potentially competing with the generation of hydroxyTDBP-TAZTO. These findings provide detailed mechanistic insight into TDBP-TAZTO biotransformation by CYP450s, which can enrich our understanding of the metabolic fate and associated health risk of this chemical.
研究领域:
环境科学
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毒理学
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计算生物学
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Plants-Basel
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2021
·Vol. 10
·期 7
·1309
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IF 3.935
Xiaoyue Yu,Hanwen Wu,Jianping Zhang,Yongjie Yang,Wei Tang,Yongliang Lu
Polypogon fugaxis a common winter weed in China and other Asia countries. We have previously found aP. fugaxbiotype (R) resistant to acetyl co-enzyme A carboxylase (ACCase) herbicides also cannot be effectively controlled by some acetolactate synthase (ALS) herbicides. This study evaluated the level of resistance to four ALS herbicides (metsulfuron-methyl, chlorsulfuron, monosulfuron, pyribambenz isopropyl) in the R biotype and the associated resistance mechanism. The R biotype exhibited moderate level of resistance to metsulfuron-methyl (6.0-fold) compared with the sensitive biotype (S). Sequence analysis ofALSgene revealed that twoALSgenes existed inP. fugax. However, no substitution associated with ALS resistance mechanism were found inALSgenes between the S and R biotypes. The activity of ALS enzyme isolated from the R biotype was inherently higher and less sensitive to metsulfuron-methyl than the S biotype. Glutathione S-transferases (GST) activity was also less sensitive to metsulfuron-methyl in the R than as the S biotypes. Malathion, a cytochrome P450 (CYP) monooxygenase inhibitor, had much greater synergistic effect with metsulfuron-methyl on the R than as the S plants, reducing the ED50value (herbicide dose to inhibit growth by 50%) of metsulfuron-methyl by 23- and 6-fold, respectively, suggesting that CYP mediated enhanced metabolism might contribute to the resistance to ALS herbicides. These results suggest that metsulfuron-methyl resistance in the R biotype was associated with the up-regulated ALS enzymatic activity and the GST and CYP-mediated enhanced herbicide metabolism.Keywords:metsulfuron-methyl;ALS enzyme activity;Polypogon fugax;glutathione S-transferases;cytochrome P450
研究领域:
植物学
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分子生物学
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农业科学
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PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
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2021
·
IF 3.963
Kai Tian,Jing Feng,Jiang Zhu,Jiagao Cheng,Mei Li,Xinghui Qiu
The cotton bollworm , Helicoverpa armigera , is a polyphagous pest threatening many economically important crops worldwide. Until recently, synthetic pyrethroids remain in wide use for controlling pest insects including the cotton bollworm. Understanding the metabolic mechanism of pyrethroids in a given pest can provide significant implication for a smart choice of insecticides, and such information is useful for the development of novel selective and safe insecticides. In this study, we used complexes of recombinant H. armigera cytochrome P450 CYP9A and NADPH-dependent cytochrome P450 reductase to investigate the capacity of three CYP9A paralogs in the transformation of seven structurally different pyrethroids by metabolism assays. The results showed that the three paralogous CYP9As were able to metabolize multiple pyrethroids. Interestingly, all the three CYP9As transformed pyrethrin-resembling pyrethroids (e.g. bioallethrin) more efficiently than the heavily modified ones (e.g. bifenthrin). These findings suggest that herbivorous insects can cope with synthetic insecticides using their physiological systems that initially evolved to survive exposure to the defensive chemicals in their host plants, adding support to the pre-adaptation hypothesis.
研究领域:
昆虫学
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农药科学
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JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
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2017
·
IF 2.603
Lingli He,Chuan Li,Xuyuan Liu,Qingqing Yang,Haizhi Zhang,Weiren Xu,Luyong Zhang,Changxiao Liu
Benzbromarone is a uricosuric drug metabolized predominantly by cytochrome P450 2C9 from in vitro findings. Human CYP2C9 exhibits extensive genetic polymorphism and numbers of clinic studies have demonstrated that CYP2C9 genetic polymorphism has a significant influence on the pharmacokinetics of benzbromarone. But in vitro study on the interaction between CYP2C9 allelic isoforms and benzbromarone was rare. Here, an LC–MS/MS method was established and validated to determine the concentration of benzbromarone in different CYP2C9 enzyme incubation systems for the drug-enzyme interaction study. By selecting appropriate internal standard and optimizing separation system, including mobile phase , sample solvent and gradient elution condition, this LC–MS/MS method was developed with fine linearity (r 2 ≥ 0.996), good reproducibility (RSD ≤ 6.6%), high stability (92.37–114.67%), efficient recovery (91.23–109.82%) and acceptable matrix effect (110.54–115.31%). Based on this method, the interaction between 3 CYP2C9 allelic isoforms and benzbromarone was researched by kinetics parameters (K m , V max , Cl int ). As a result, CYP2C9*1 displayed the highest metabolic activity towards benzbromarone, CYP2C9*2 showed a little lower catalytic activity than CYP2C9*1 (relative clearance/*1 = 85.86%), CYP2C9*3 showed the lowest catalytic activity (relative clearance/*1 = 21.57%). The result illustrated that various CYP2C9 allelic isoforms showed different enzymatic activities towards benzbromarone, which could offer effective consultation for personalized administration in clinic.
研究领域:
药理学
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医学遗传学
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危险品化学品经营许可证（带存储）