PEST MANAGEMENT SCIENCE
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2022
·Vol. 79
·期 4
·1452-1466
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IF 4.462
Pei-Rong Li,Yu Shi,Di Ju,Yu-Xi Liu,Wei Wang,Ying-Shi He,Yu-Yun Zhang,Xue-Qing Yang
BACKGROUND The evolution of insect resistance to pesticides poses a continuing threat to sustainable pest management. While much is known about the molecular mechanisms that confer resistance in model insects and few agricultural pests, far less is known about fruit pests. Field-evolved resistance to synthetic insecticides such as lambda -cyhalothrin has been widely documented in Cydia pomonella , a major invasive pest of pome fruit worldwide, and the increased production of cytochrome P450 monooxygenases (P450s) has been linked to resistance in field-evolved resistant populations. However, the underlying molecular mechanisms of P450-mediated insecticide resistance remain largely unknown . RESULTS Here we found that functional redundancy and preference of metabolism by P450s genes in the CYP9A subfamily confer resistance to lambda -cyhalothrin in Cydia pomonella . A total of four CYP9A genes, including CYP9A61 , CYP9A120 , CYP9A121 , and CYP9A122 , were identified from Cydia pomonella . Among these, CYP9A120 , CYP9A121 , and CYP9A122 were predominantly expressed in the midgut of larvae. The expression levels of these P450 genes were significantly induced by a lethal dose that would kill 10% (LD 10 ) of lambda -cyhalothrin and were overexpressed in a field-evolved lambda -cyhalothrin resistant population. Knockdown of CYP9A120 and CYP9A121 by RNA-mediated interference (RNAi) increased the susceptibility of larvae to lambda -cyhalothrin. In vitro assays demonstrated that recombinant P450s expressed in Sf9 cells can metabolize lambda -cyhalothrin, but with functional redundancy and divergence through regioselectivity of metabolism. CYP9A121 preferred to convert lambda -cyhalothrin to 2′-hydroxy- lambda -cyhalothrin, whereas CYP9A122 only generated 4′-hydroxy metabolite of lambda -cyhalothrin. Although possesses a relatively low metabolic capability, CYP9A120 balanced catalytic competence to generate both 2′- and 4′-metabolites. CONCLUSION Collectively, these results reveal that metabolic functional redundancy of three members of the CYP9A subfamily leads to P450-mediated lambda -cyhalothrin resistance in Cydia pomonella , thus representing a potential adaptive evolutionary strategy during its worldwide expansion. © 2022 Society of Chemical Industry.
研究领域:
昆虫学
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分子生物学
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杀虫剂抗性
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DOI
PubMed
高影响因子
ENVIRONMENTAL RESEARCH
·
2025
·
IF 7.7
Lingmin Jin,Yuchen Lu,Jiawei Huang,Jiangmin Liu,Xiaoxuan Wei,Guangcai Ma,Haiying Yu
The nitrogen-contained disinfection by-products, halonitromethanes (HNMs), are known for their high cytotoxicity and genotoxicity. Although HNMs can be metabolized by cytochrome P450 enzymes (P450s), the specific mechanism has remained unclear. To shed light on this, density functional theory (DFT) calculations were performed to elucidate the potential oxidative P450-catalytic activation of the nine HNMs. Our findings reveal that active species of P450s (Cpd I) predominantly react with halogen-substituted nitromethanes via hydrogen abstraction and bromine atom abstraction, rather than chlorosylation. As a result of these reactions, oxidized HNMs are produced and can undergo further hydrolysis, leading to nitro-formaldehyde, nitro formyl halogen, halogen hydride, hypobromous acid, and nitroformic acid. To experimentally validate the computational predictions, in vitro experiments were conducted on five typical nitromethanes using human liver microsomes and the results reveal that DCNM, BCNM and DBCNM form nitroformyl chlorine (NO 2 CClO), while BCNM, DBNM and TBNM are transferred into nitroformyl bromide (NO 2 CBrO). Nitroformic acid is also identified as a metabolite in the TBNM metabolism reaction. Toxicity assessment reveals that metabolic transformation leads to an overall reduction in the ecological toxicity. However, metabolites showed similar toxicity to Fathead minnow and even higher acute toxicity to rat, as well as larger probability of hERG inhibition effects than HNMs, underscoring the need for caution in health risk assessment. By integrating in silico and in vitro approaches, this work has provided a comprehensive understanding of the metabolism of HNMs and offered potential toxicity data basis of these compounds.
研究领域:
环境毒理学
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生物化学
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计算化学
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药理学
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水处理化学
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DOI
PubMed
高影响因子
XENOBIOTICA
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2023
·
IF 1.8
Fengping Chen,Haizhu Zou,Ping Zhang,Yuqi Yan
In vitro metabolism of bicyclol was studied using liver microsomes, hepatocytes and human recombinant cytochrome P450 enzymes. Liquid chromatography-benchtop orbitrap mass spectrometry technique was utilised to identify the metabolites.A total of 19 metabolites, including 5 new metabolites (M2, M3, M4, M5 and M16) were tentatively identified. Among these metabolites, M6&M8 (demethylenation), M9&M10 (demethylation) and M19 (glucuronidation) were the major metabolites.In glutathione (GSH)-supplemented liver microsomes, 5 new GSH conjugates were found and tentatively identified. The formation was assumed to be through demethylenation of methylenedioxyphenyl to form catechol derivatives, which further underwent oxidation to form ortho-quinone intermediates, reacting with GSH to form stable adducts.CYP3A4 and 2C19 were demonstrated to be the major enzymes responsible for the bioactivation of bicyclol.This study provided valuable information on the metabolic fate of bicyclol in liver microsomes and hepatocytes, and the bioactivation pathways were reported for the first time, which would be helpful for us to understand the potential drug-drug interactions and the possible side effect of this drug.
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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DOI
PubMed
高影响因子
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
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2023
·Vol. 89
·期 3
·
IF 4.4
Ling Wang,Xiaowei Wu,Chenghua Gao,Lingrui Wei,Qian Li,Aitao Li
In this study, a new cytochrome P450 enzyme, namely, CYP68J5_Fusarium graminearum (CYP68J5_fg), was identified from Fusarium graminearum via a combination of transcriptome sequencing and heterologous expression in Saccharomyces cerevisiae. The biotransformation of progesterone by whole-cells of S. cerevisiae expressing CYP68J5_fg revealed that the CYP68J5_fg possessed steroidal 12β- and 15α-hydroxylase activities. To the best of our knowledge, this is the first time that a fungal P450 enzyme with 12β-hydroxylase activity has been identified. This advance offers opportunities to boost the efficiency and selectivity of the CYP68J5_fg hydroxylating system and thus shows great potential for further applications of this enzyme for the synthesis of steroid drugs.IMPORTANCE Regioselective and stereoselective hydroxylation is of vital importance in the functionalization of steroids, which remains challenging in organic synthesis. In particular, the C12-hydroxy steroids play a significant role in the synthesis of many important steroidal drugs. In this study, a novel fungal P450 enzyme with 12β-hydroxylation activity was identified, and it shows different substrate specificity and regioselectivity, compared to the bacterial and fungal steroidal hydroxylases that are known to date. This lays the foundation for the creation of effective biocatalysts for the process of 12β-hydroxylation, although further understanding of the molecular structural basis of this fungal P450 is needed to facilitate the engineering of this enzyme for industrial applications.
研究领域:
生物化学
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酶学
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药学
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微生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2022
·Vol. 23
·期 19
·11578
·
IF 6.208
Bin Liu,Ming Tang,Hui Chen
Bark beetles mainly rely on detoxification enzymes to resist the host tree’s defense against oleoresin terpenes. Cytochrome P450 enzymes (CYPs) play an important role in the detoxification of plant allelochemicals and pesticides in insect. One P450 gene (DaCYP4BQ1) is associated with the response of (+)-α-pinene inDendroctonus armandi. However, the regulatory mechanism of this P450 gene response to (+)-α-pinene is still unknown. In this study, spatiotemporal expression profiling indicated thatCYP4BQ1was highly expressed in adult and larval stages ofD. armandi, and it was predominantly expressed in fat body, midgut, and Malpighian tubules of adults. Moreover, the expression ofCYP4BQ1significantly increased after exposure to (+)-α-pinene, and depletion of it decreased the tolerance of adults to (+)-α-pinene. In addition, (+)-α-pinene treatment induced the expression of the transcription factors cap ‘n’ collar isoform C (CncC) and its binding factor muscle aponeurosis fibromatosis (Maf), elevated the level of hydrogen peroxide (H2O2), and increased the activities of antioxidant enzymes. SilencingCncCsuppressedCYP4BQ1expression and enhanced the susceptibility of beetles to (+)-α-pinene. Similarly, application of the reactive oxygen species (ROS) scavenger N-acetylcysteine reduced the production and accumulation of H2O2, suppressed the expression ofCncC, Maf, andCYP4BQ1and led to decreased tolerance of adults to (+)-α-pinene. In contrast, ingestion of the CncC agonist curcumin elevatedCYP4BQ1expression and enhanced (+)-α-pinene tolerance. The results demonstrate that, inD. armandi, (+)-α-pinene inducesCYP4BQ1via activation of the ROS/CncC signaling pathway.Keywords:Dendroctonus armandi;phytochemical tolerance;cytochrome P450;oxidative burst;CncC pathway
研究领域:
昆虫学
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生物化学
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分子生物学
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DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
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2023
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IF 15.1
Xiang Wang,Zhong-Hong Zhang,Kuan-Kuan Yuan,Hui-Ying Xu,Guo-Hui He,Libin Yang,Joseph Buhagiar,Wei-Dong Yang,Yalei Zhang,Carol Sze Ki Lin,Hong-Ye Li
Microalgae-based antibiotic removal treatment has attracted attention because of its low carbon and sustainable advantages. The microalgal co-metabolism system with a suitable carbon source leads to enhanced performance of pollutant removal. However, currently, limited knowledge is available for the removal of fluoroquinolone using a microalgae-mediated co-metabolism system. In this study, we first investigated that the biotic processes by alga Haematococcus lacustris in the co-metabolism system by adding glycerol would be the main contributors responsible for the removal of 10 mg/L ofloxacin (OFL) with the efficiency of 79.73% and the removal of 10 mg/L enrofloxacin (ENR) with the efficiency of 54.10%, respectively. Furthermore, we found that pyruvate from glycerol was converted into substrates and precursors, thereby resulting in the significant accumulations of microalgal astaxanthin and lipid. The astaxanthin content of H. lacustris was achieved at 4.81% and 4.69% treated with OFL and ENR in the presence of glycerol, with 16.04% and 14.55% of lipid content, respectively. The proposed metabolites and pathways were identified to plausibly explain the biodegradation of fluoroquinolone by H. lacustris . The molecular analyses demonstrated that cytochrome P450 (CYP450) enzymes are responsible for the biodegradation of fluoroquinolone, and it was further verified that fluoroquinolones might insert into CYP450 to finally form an efficient and tight binding conformation by molecular dynamic simulation. These findings provide a microalgae-based route for feasible and sustainable biodegradation of antibiotics using a co-metabolism strategy comprising glycerol as a carbon source, with the synergistic accumulation of valuable products.
研究领域:
环境科学
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微生物学
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生物技术
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药理学
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产品关联
DOI
高影响因子
Frontiers in Pharmacology
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2021
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IF 5.811
Wang Jinhui,Chen Feifei,Jiang Hui,Xu Jia,Meng Deru,Geng Peiwu,Dai Dapeng,Hu Jingbo,Zhou Yunfang,Zhou Quan,Wang Shuanghu
Poziotinib is an orally active, irreversible, pan-HER tyrosine kinase inhibitor used to treat non-small cell lung cancer, breast cancer, and gastric cancer. Poziotinib is currently under clinical investigation, and understanding its drug-drug interactions is extremely important for its future development and clinical application. The cocktail method is most suitable for evaluating the activity of cytochrome P450 enzymes (CYPs). As poziotinib is partially metabolized by CYPs, cocktail probes are used to study the interaction between drugs metabolized by each CYP subtype. Midazolam, bupropion, dextromethorphan, tolbutamide, chlorzoxazone, phenacetin, and their metabolites were used to examine the effects of poziotinib on the activity of cyp1a2, 2b1, 2d1, 2c11, 2e1, and 3a1/2, respectively. The in vitro experiment was carried out by using rat liver microsomes (RLMs), whereas the in vivo experiment involved the comparison of the pharmacokinetic parameters of the probes after co-administration with poziotinib to rats to those of control rats treated with only probes. UPLC-MS/MS was used to detect the probes and their metabolites in rat plasma and rat liver microsomes. The in vitro results revealed that the half-maximal inhibitory concentration values of bupropion and tolbutamide in RLMs were 8.79 and 20.17 μM, respectively, indicating that poziotinib showed varying degrees of inhibition toward cyp2b1 and cyp2c11. Poziotinib was a competitive inhibitor of cyp2b1 and cyp2c11, with Ki values of 16.18 and 17.66 μM, respectively. No time- or concentration-dependence of inhibition by poziotinib was observed toward cyp2b1 and cyp2c11 in RLMs. Additionally, no obvious inhibitory effects were observed on the activity of cyp1a2, cyp2d1, cyp2e1, and cyp3a1/2. In vivo analysis revealed that bupropion, tolbutamide, phenacetin, and chlorzoxazone showed significantly different pharmacokinetic parameters after administration (p < 0.05); there was no significant difference in the pharmacokinetic parameters of dextromethorphan and midazolam. These results show that poziotinib inhibited cyp2b1 and cyp2c11, but induced cyp1a2 and cyp2e1 in rats. Thus, poziotinib inhibited cyp2b1 and cyp2c11 activity in rats, suggesting the possibility of interactions between poziotinib and these CYP substrates and the need for caution when combining them in clinical settings.
研究领域:
药理学
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临床医学
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生物化学
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DOI
高影响因子
CHEMICAL RESEARCH IN TOXICOLOGY
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2022
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IF 3.973
Min Zhao,Junzu Shi,Wei Li,Chunjing Guan,Chen Sun,Ying Peng,Jiang Zheng
Gemfibrozil (GEM), a lipid regulator, is a fibric acid derivative widely used in the treatment of hyperlipidemia. It has been reported that GEM can induce acute liver injury in the course of therapy in clinical practice, so it is necessary to elucidate the mechanisms of toxic action. The present study focused on metabolic activation of GEM, possibly participating in GEM-mediated liver injury. A benzylic alcohol metabolite (M1), along with a phenol metabolite (M2), was detected in microsomal incubations, rat primary hepatocyte culturing, and rats given GEM. A GSH conjugate (M3) was detected in cultured rat hepatocytes after exposure to GEM. Formation of M1 was found to be NADPH dependent, and generation of M3 required M1 and 3′-phosphoadenosine-5′-phosphosulfate. It is most likely that GEM was biotransformed to M1, which was further metabolized to a sulfate. The resulting sulfate was reactive to bio-thiols. Cytochrome P450 and sulfotransferases participated in the phase I and phase II reactions, respectively. M1 and M3 were chemically synthesized, and their structures were characterized by mass spectrometry and NMR. The present study has particular value for elucidating the mechanism of liver injury caused by GEM.
研究领域:
药理学
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毒理学
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药物代谢
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DOI
PubMed
高影响因子
TOXICOLOGY AND APPLIED PHARMACOLOGY
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2017
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IF 3.791
Jun-Jun Chen,Jing-Xian Zhang,Xiang-Qi Zhang,Mei-Juan Qi,Mei-Zhi Shi,Jiao Yang,Ke-Zhi Zhang,Cheng Guo,Yong-Long Han
Diosmetin (3′, 5, 7-trihydroxy-4′-methoxyflavone), a natural flavonoid from traditional Chinese herbs, has been used in various medicinal products because of its anticancer, antimicrobial, antioxidant, estrogenic and anti-inflammatory activity. However, flavonoids could affect the metabolic enzymes and cause drug-drug interactions (DDI), reducing the efficacy of co-administered drugs and potentially resulting in serious adverse reactions. To evaluate its potential to interact with co-administered drugs, the IC 50 value of phase I cytochrome P450 enzymes (CYPs), phase II UDP-glucuronyltransferases (UGTs) and hepatic uptake transporters (organic cation transporters (OCTs), organic anion transporter polypeptides (OATPs) and Na + -taurocholate cotransporting polypeptides (NTCPs)) were examined in vitro by LC-MS/MS. Diosmetin showed strong inhibition of CYP1A2 in a concentration-dependent manner. The intensity of the inhibitory effect was followed by CYP2C8 , CYP2C9 , CYP2C19 and CYP2E1 . For CYP2A6 , CYP2B6 , CYP2D6 and CYP3A4 , diosmetin was found to have no significant inhibitory effects, and the induction effect on CYPs was not significant. For UGTs, diosmetin had a minimal inhibitory effect. In addition, the inhibitory effects of diosmetin on OATP and OCT1 were weak, and it had little effect on NTCP. This finding indicated that drug-drug interactions induced by diosmetin may occur through co-administration of drugs metabolized by CYP1A2.
研究领域:
药理学
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毒理学
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生物化学
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DOI
PubMed
高影响因子
Frontiers in Chemistry
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2021
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IF 5.221
Zhang Xiaodong,Hu Yun,Peng Wei,Gao Chenghua,Xing Qiong,Wang Binju,Li Aitao
Cytochrome P450 enzyme CYP109B1 is a versatile biocatalyst exhibiting hydroxylation activities toward various substrates. However, the regio- and stereoselective steroid hydroxylation by CYP109B1 is far less explored. In this study, the oxidizing activity of CYP109B1 is reconstituted by coupling redox pairs from different sources, or by fusing it to the reductase domain of two self-sufficient P450 enzymes P450RhF and P450BM3 to generate the fused enzyme. The recombinant Escherichia coli expressing necessary proteins are individually constructed and compared in steroid hydroxylation. The ferredoxin reductase (Fdr_0978) and ferredoxin (Fdx_1499) from Synechococcus elongates is found to be the best redox pair for CYP109B1, which gives above 99% conversion with 73% 15β selectivity for testosterone. By contrast, the rest ones and the fused enzymes show much less or negligible activity. With the aid of redox pair of Fdr_0978/Fdx_1499, CYP109B1 is used for hydroxylating different steroids. The results show that CYP109B1 displayed good to excellent activity and selectivity toward four testosterone derivatives, giving all 15β-hydroxylated steroids as main products except for 9 (10)-dehydronandrolone, for which the selectivity is shifted to 16β. While for substrates bearing bulky substitutions at C17 position, the activity is essentially lost. Finally, the origin of activity and selectivity for CYP109B1 catalyzed steroid hydroxylation is revealed by computational analysis, thus providing theoretical basis for directed evolution to further improve its catalytic properties.
研究领域:
生物化学
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生物催化
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酶学
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类固醇化学
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DOI
PubMed
高影响因子
Nature Communications
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2020
·Vol. 11
·期 1
·1-12
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IF 12.121
Tu Lichan,Su Ping,Zhang Zhongren,Gao Linhui,Wang Jiadian,Hu Tianyuan,Zhou Jiawei,Zhang Yifeng,Zhao Yujun,Liu Yuan,Song Yadi,Tong Yuru,Lu Yun,Yang Jian,Xu Cao,Jia Meirong,Peters Reuben J.,Huang Luqi,Gao Wei
Triptolide is a trace natural product of Tripterygium wilfordii . It has antitumor activities, particularly against pancreatic cancer cells. Identification of genes and elucidation of the biosynthetic pathway leading to triptolide are the prerequisite for heterologous bioproduction. Here, we report a reference-grade genome of T. wilfordii with a contig N50 of 4.36 Mb. We show that copy numbers of triptolide biosynthetic pathway genes are impacted by a recent whole-genome triplication event. We further integrate genomic, transcriptomic, and metabolomic data to map a gene-to-metabolite network. This leads to the identification of a cytochrome P450 (CYP728B70) that can catalyze oxidation of a methyl to the acid moiety of dehydroabietic acid in triptolide biosynthesis. We think the genomic resource and the candidate genes reported here set the foundation to fully reveal triptolide biosynthetic pathway and consequently the heterologous bioproduction.
研究领域:
植物生物学
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基因组学
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药理学
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分子生物学
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DOI
PubMed
高影响因子
BMC BIOLOGY
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2024
·Vol. 22
·期 1
·1-14
·
IF 4.4
Jia Qiangqiang,Yang Liu,Wen Jiamin,Liu Suning,Wen Di,Luo Wei,Wang Weihua,Palli Subba Reddy,Sheng Li
Background Juvenile hormones (JH) play crucial role in regulating development and reproduction in insects. The most common form of JH is JH III, derived from MF through epoxidation by CYP15 enzymes. However, in the higher dipterans, such as the fruitfly, Drosophila melanogaster , a bis-epoxide form of JHB3, accounted most of the JH detected. Moreover, these higher dipterans have lost the CYP15 gene from their genomes. As a result, the identity of the P450 epoxidase in the JH biosynthesis pathway in higher dipterans remains unknown. Results In this study, we show that Cyp6g2 serves as the major JH epoxidase responsible for the biosynthesis of JHB3 and JH III in D. melanogaster . The Cyp6g2 is predominantly expressed in the corpus allatum (CA), concurring with the expression pattern of jhamt , another well-studied gene that is crucial in the last steps of JH biosynthesis. Mutation in Cyp6g2 leads to severe disruptions in larval-pupal metamorphosis and exhibits reproductive deficiencies, exceeding those seen in jhamt mutants. Notably, Cyp6g2 −/− ::jhamt 2 double mutants all died at the pupal stage but could be rescued through the topical application of JH analogs. JH titer analyses revealed that both Cyp6g2 −/− mutant and jhamt 2 mutant lacking JHB3 and JH III, while overexpression of Cyp6g2 or jhamt caused a significant increase in JHB3 and JH III titer. Conclusions These findings collectively established that Cyp6g2 as the major JH epoxidase in the higher dipterans and laid the groundwork for the further understanding of JH biosynthesis. Moreover, these findings pave the way for developing specific Cyp6g2 inhibitors as insect growth regulators or insecticides.
研究领域:
昆虫学
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分子生物学
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发育生物学
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遗传学
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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）