BIOMEDICAL CHROMATOGRAPHY
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2021
·Vol. 36
·期 1
·e5239
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IF 1.902
Xiao-fang Ma,Qi Zhao,Yan Cheng,Dong-Mei Yan,Wei-Feng Zhu,Fei Li
Coumarins are a group of natural compounds commonly found in the families of Rutaceae and Umbelliferae. 7-Isopentenyloxycoumarin (ISC), auraptene (AUR), and umbelliprenin (UM) belong to prenyloxycoumarins (PYCs), which link isopentenyl, geranyl, and farnesyl group at C7 position, respectively. The substituent of 7-ethoxycoumarin (ETC) is the ethyl group. In this study, UPLC–ESI–QTOF–MS (ultra-performance liquid chromatography–electrospray ionization–quadrupole time of flight–MS)-based metabolomics was used to evaluate the in vivo and in vitro metabolism of PYCs. Results showed that ETC produced 10 known metabolites, and ISC was transformed into 17 metabolites in vivo and in vitro , which were undescribed compounds. A total of 35 AUR metabolites, including 34 undescribed metabolites were identified, and 21 metabolites were reported for the first time in UM. The results indicated that hydroxylation and N -acetylcysteine conjugation were the common metabolic reactions for PYCs. The metabolic rates of ETC, ISC, AUR and UM were 26%, 36%, 81%, and 38%, respectively, in human liver microsome, while they were 24%, 40%, 80%, and 37%, respectively, in mouse liver microsomes. In addition, recombinant cytochrome P450s (CYPs) screening showed that CYP1A1, 2C19, 3A4, and 3A5 were the major metabolic enzymes involved in the formation of hydroxylation metabolites. Together, these results suggest that the isopentenyl group plays an important role in the metabolism of PYCs.
研究领域:
药理学
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植物化学
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代谢研究
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PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2023
·
IF 4.7
Tian-yi Zhang,Chang-wei Gong,Jian Pu,An-chun Peng,Xu-yang Li,Yu-meng Wang,Xue-gui Wang
Solenopsis invicta is a main issue in southern China and is causing significant damage to the local ecological environment. The extensive use of insecticides has resulted in the development of tolerance in S. invicta . In our study, ten S. invicta colonies from Sichuan Province exhibited varying degrees of tolerance against flonicamid, with LC 50 values from 0.49 mg/L to 8.54 mg/L. The sensitivity of S. invicta to flonicamid significantly increased after treatment with the P450 enzyme inhibitor piperonyl butoxide (PBO). Additionally, the activity of P450 in S. invicta was significantly enhanced after being treated with flonicamid. Flonicamid induced the expression levels of CYP4aa1 , CYP9e2 , CYP4C1 , and CYP6A14 . The expression levels of these P450 genes were significantly higher in the tolerant colonies compared to the sensitive colonies, and the relative copy numbers of CYP6A14 in the tolerant colonies were 2.01–2.15 fold. RNAi feeding treatment effectively inhibited the expression of P450 genes, thereby reducing the tolerance of S. invicta against flonicamid. In addition, the overexpression of CYP6A14 in D. melanogaster resulted in reduced sensitivity to flonicamid. Our investigations revealed hydrophobic interactions between flonicamid and seven amino acid residues of CYP6A14, along with the formation of a hydrogen bond between Glu306 and flonicamid. Our findings suggest that flonicamid can effectively control S. invicta and P450 plays a pivotal role in the tolerance of S. invicta against flonicamid. The overexpression of CYP6A14 also increased tolerance to flonicamid.
研究领域:
昆虫学
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分子生物学
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毒理学
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Synthetic and Systems Biotechnology
·
2026
·
IF 4.8
Jian Yang,Rong Li,Qilin Gao,Li Ma,Qiang Wang,Guiying Ma,Yikang Fan,Shengying Li,Lian-Hua Xu
C21-hydroxylation is a crucial step in the synthesis of corticosteroids. For instance, C21-hydroxylase catalyzes the conversion of progesterone (PRO) into 11-deoxycorticosterone (DOC, the precursor of cortisol and aldosterone). Through structure-guided rational design combined with focused rational iterative site-directed mutagenesis (FRISM), we engineered the wild-type CYP154C5 from Nocardia farcinica to obtain the highly efficient variant M6a (F92A/V291L/L294I/Q239K/F180W/Q398M). This variant demonstrated a complete regioselectivity shift from C16α to C21 hydroxylation (98% selectivity) of PRO with high catalytic efficiency (>99% conversion). To further enhance catalytic performance, we engineered a redox fusion variant (M6a-RhFRED L3) that demonstrated the highest catalytic activity, achieving a 1.43-fold improvement compared to the M6a-RhFRED. Additionally, M6a-RhFRED L3 catalyzed the C21-hydroxylation of three steroid analogs (dydrogesterone, 16-dehydroprogesterone, and pregna-4,6-diene-3,20-dione), with selectivity reaching 98% and conversion rates exceeding 60%. Molecular docking and molecular dynamics (MD) simulations revealed that the PRO substrate undergoes conformational rearrangement in the M6a active site. This structural reorganization underscores the critical role of key residues in modulating regioselectivity, particularly the shift from C16α to C21 hydroxylation. This study not only provides an efficient biocatalyst for steroids C21-hydroxylation but also offers valuable insight for the rational engineering of the P450 enzymes.
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CHEMICO-BIOLOGICAL INTERACTIONS
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2026
·
IF 5.2
Yiting You,Pengxiang Wang,Jingqian Huang,Xiaoyong Zhang,Feng Han,Jun Liu,Danhong Xu,Yujian Liu,Xiaochen Bao,Xiaoyan Zhu
Although hyperbaric oxygen (HBO) therapy is a clinically established intervention for various pathological conditions, excessive oxygen exposure can induce acute lung injury (ALI), the underlying mechanisms of which remain elusive. In this study, we demonstrate that neutrophil extracellular traps (NETs) contribute to the pathogenesis of HBO-induced ALI. HBO exposure upregulates pulmonary expression of CYP1A1 and CYP1A2, two isoforms of the cytochrome P450 (CYP) 1A subfamily. Genetic deletion of both CYP1A1 and CYP1A2 exacerbates, whereas the CYP1A inducer attenuates NET formation and ALI in HBO-exposed mice. Mechanistically, HBO-induced upregulation of CYP1A protects against lung injury by suppressing oxidative stress-driven NET formation via an NRF2(nuclear factor erythroid 2-related factor 2)-dependent pathway. Notably, the inhibitory effect of the CYP1A/NRF2 signaling axis on NET formation is mediated indirectly through its antioxidant actions in alveolar epithelial cells, rather than via direct modulation of neutrophils, thereby establishing an antioxidative microenvironment that limits NET formation. Our findings reveal a novel protective mechanism whereby activation of the CYP1A/NRF2 pathway mitigates HBO-induced lung injury through regulation of intercellular crosstalk between alveolar epithelial cells and neutrophils within the oxidative milieu.
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PLOS BIOLOGY
·
2026
·Vol. 24
·期 9
·e3004013
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IF 6.9
Huidong Wang,Yajie Kong,Xinyue Su,Yu Shi,Jianpeng Zhang,Yinjia Wang,Yajuan Xiao,Xing Geng,Jing Song,Yueru Ye,Kairan Zuo,Chris Bass,Shutang Zhou
Plants and herbivorous insects are engaged in a continuous evolutionary arms race driven by chemical defences and counter-defences. How insects integrate distinct detoxification systems to overcome diverse phytochemicals, and how such mechanisms contribute to host-range divergence, remains poorly understood. Here, we uncover a cooperative cytochrome P450–UDP-glycosyltransferase (UGT) cascade that mediates furanocoumarin tolerance in Helicoverpa armigera . Comparative genomic analyses across Helicoverpa and related noctuid species revealed that the UGT33 family is the most extensively expanded and dynamically diversified UGT lineage in Helicoverpa . Targeted CRISPR–Cas9 knockouts of UGT33 gene clusters in polyphagous H. armigera demonstrated their essential roles in detoxifying the furanocoumarins xanthotoxin and imperatorin. Combined metabolic and functional assays further established a sequential detoxification pathway, in which the cytochrome P450 CYP6AE19 catalyses either the O-dealkylation of xanthotoxin to yield xanthotoxol or the aromatic-carbon hydroxylation of xanthotoxin to form 5-hydroxyxanthotoxin, which are subsequently glycosylated by UGT33 enzymes to form their less-toxic glucosides. In contrast, impaired CYP6AE19–UGT33 coordination in the oligophagous Helicoverpa assulta was associated with reduced xanthotoxin detoxification and high sensitivity to this compound. Together, these findings provide direct evidence for coordinated Phase I–Phase II detoxification of a plant defensive compound in insects and show that functional divergence in this pathway contributes to interspecific differences in plant toxin tolerance, which are associated with contrasting dietary breadth in closely related herbivorous insects.
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ACS Catalysis
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2026
·
IF 13.6
Jun-Bin He,Lian Wu,Wenli Yuan,Hai-Xue Pan,Binju Wang,Gong-Li Tang
Achieving highly regioselective C–H bond functionalization at the bridgehead carbon of a single molecule is a fascinating, yet formidable challenge. BcmD, a P450 enzyme, catalyzes the regioselective hydroxylation at the sp3-hybridized bridgehead carbon of the bicyclo[4.2.2]piperazinedione ring scaffold during bicyclomycin biosynthesis. Herein, we provide mechanistic details of BcmD to understand its substrate binding and the origin of its regioselectivity. The high-resolution structure of SoBcmD from Streptomyces ossamyceticus in complex with its natural substrate, together with site-directed mutagenesis and computational simulations, reveals that the precise substrate positioning, enabled by specific hydrogen-bonding and hydrophobic interactions with crucial residues, determines the regioselectivity. Intriguingly, we identify SoBcmD as a peroxygenase that could utilize H2O2 as an oxidant to catalyze regioselective hydroxylation. Mechanistic analysis indicates that H2O2 activation in SoBcmD involves a homolytic cleavage process, mediated by a hydrogen-bonding network comprising residues T277, A273, and the substrate. Furthermore, structure-based channel analysis and mutagenesis demonstrate that the I157V mutant exhibits significantly enhanced peroxygenase activity. These findings advance our understanding of regioselectivity control in BcmD and provide valuable guidance for engineering P450 peroxygenases for the C–H bond activation at the chemically challenging bridgehead carbon sites.
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Journal of Integrative Agriculture
·
2026
·
IF 4.4
Xiaolan Liu,Zhuang Zhang,Kaixin Li,Zanrong Wen,Wei Xu,Huiwen Tan,Xichao Hu,Lei Guo
The whitefly, Bemisia tabaci , is a major agricultural pest that has developed resistance to a broad range of insecticides. Despite the promising efficacy of cyantraniliprole (CYA) against B . tabaci , medium to high levels of resistance have emerged after prolonged field use. However, the mechanisms driving CYA resistance remain poorly understood. In this study, four B . tabaci strains exhibiting 24.9-fold to 28.9-fold resistance ratio to CYA were investigated. Synergist assays and enzyme activity measurements indicated cytochrome P450 enzymes contribute to this resistance. RNA sequencing and RT-qPCR analysis identified five P450 genes ( CYP305H2, CYP6EM1, CYP3133D3, CYP3133D5 , and CYP3133E2 ) as significantly overexpressed in resistant strains. Targeted silencing of these genes led to a 22.3% to 50.3% increase in CYA toxicity. The metabolic rates of these P450 enzymes against CYA were 2.5- to 6.0-fold higher than that in the control group within two hours. These results provide new insights into the molecular basis of CYA resistance in B . tabaci and highlight the pivotal role of cytochrome P450 enzymes in metabolic adaptation to diamide insecticides.
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Theranostics
·
2020
·
IF 8.579
Qiu Wang,Mengchi Sun,Dan Li,Chang Li,Cong Luo,Zhaomeng Wang,Wenjuan Zhang,Zimeng Yang,Yao Feng,Shuang Wang,Zhonggui He,Haotian Zhang,Qiming Kan,Wei Sun,Jin Sun
Reactive oxygen species (ROS)-based photodynamic therapy (PDT) has a widespread application in cancer therapy. Nevertheless, the efficiency of PDT is far from satisfactory. One major impediment is the overexpression of glutathione (GSH) in tumor cells, which could deplete the level of PDT-generated ROS. Herein, we develop a novel type of cytochrome P450 enzyme-mediated auto-enhanced photodynamic co-nanoassembly between clopidogrel (CPG) and photosensitizer pyropheophorbide a (PPa). Methods: In this work, we prepare the co-assembled nanoparticles of CPG and PPa (CPG/PPa NPs) by using one-step precipitation method. The assembly mechanism, drug release behavior, GSH consumption, ROS generation, cellular uptake, cytotoxicity of CPG/PPa NPs are investigated in vitro . The mice bearing 4T1 tumor are employed to evaluate in vivo biodistribution and anti-tumor effect of CPG/PPa NPs. Results: Such CPG/PPa NPs could disrupt the intracellular redox homeostasis, resulting from the elimination of GSH by CPG active metabolite mediated by cytochrome P450 enzyme (CYP2C19). The in vivo assays reveal that CPG/PPa NPs not only increase the drug accumulation in tumor sites but also significantly suppress tumor growth in BALB/c mice bearing 4T1 tumor. With CPG-mediated GSH consumption and PPa-triggered ROS generation, CPG/PPa NPs show the enhanced PDT treatment effect by breaking intracellular redox balance. Conclusion: Our findings provide a valuable knowledge for the rational design of the PDT-based combinational cancer therapy.
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PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY
·
2025
·
IF 4
Fang-Rui Guo,Shu-Chao Wang,Yan Liu,Shuai Wang,Jing-Mei Huang,Hao Sun,Lin-Feng He,Yuan Xie,Song-Tao Qiao,Feng-Xia Yang,Chris Bass,Cong-Fen Gao,Shun-Fan Wu
The development of insecticide resistance in insect populations is a major challenge to sustainable agriculture and food security worldwide. Methoxyfenozide, an insect growth regulator that acts as an agonist of 20-hydroxyecdysone (20E), has severely declined in its efficacy against the rice stem borer ( Chilo suppressalis ), a notorious pest of rice crops in East and Southeast Asia. To date, however, the genes involved in methoxyfenozide resistance in target pests remain unclear. We conducted a long-term (seven years from 2017 to 2023) and large geographical scale (8 provinces and 45 cities in China) resistance monitoring program for methoxyfenozide in C. suppressalis . Resistance was seen to arise rapidly in this species, with >100-fold resistance being detected in nearly all the field populations after 2018 . Piperonyl butoxide (PBO), an inhibitor of cytochrome P450 enzymes (P450s), significantly increased the sensitivity of resistant strains of C. suppressalis to methoxyfenozide, implicating P450s in resistance. Six P450 genes: CYP321F3, CYP6CV5, CYP9A68 , CYP6AB45, CYP324A12 and CYP6SN2 were identified as highly expressed in resistant C. suppressalis by transcriptome profiling. Of these, ectopic expression of CYP321F3 in Drosophila melanogaster resulted in a 7.0-fold increase in resistance to methoxyfenozide demonstrating its causal role in resistance. Collectively, these findings provide insight into the mechanisms mediating resistance to insect growth regulators and will inform the development of future pest and resistance management strategies.
研究领域:
昆虫学
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分子生物学
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农业科学
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INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
·
2015
·
IF 3.45
Lulu Dai,Mingyuan Ma,Chunyan Wang,Qi Shi,Ranran Zhang,Hui Chen
Bark beetles oxidize the defensive allelochemicals from their host trees to both detoxify and convert these materials into components of their pheromone system. The ability of this insect to adapt to its chemical environment might be explained by the action of major detoxification enzymes such as cytochrome P450s (or CYPs). Sixty-four sequences coding for P450s were identified, and most of the transcripts were found to be expressed in the larvae, pupae and adults of Dendroctonus armandi . To gain information on how these genes help D. armandi overcome the host defense, differential transcript levels of the CYP genes were observed between sexes and within the sexes. Significant differences were observed among developmental stages, in feeding on the phloem of Pinus armandi and in exposure to stimuli ((±)-α-pinene, (S)-(−)-α-pinene, (S)-(−)-β-pinene, (+)-3-carene, (±)-limonene and turpentine oil) for 8 h. We investigated the effect of sex and generations on the survivorship of individual D. armandi that were exposed to host volatiles at concentrations comparable to constitutive and induced levels of defense using fumigant exposure to understand the ability of the beetles to tolerate host defensive chemicals. The differential transcript accumulation patterns of CYP genes of these bark beetle provided insight into the ecological interactions of D. armandi with its host pine.
研究领域:
昆虫学
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分子生物学
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高影响因子
Nature Chemical Biology
·
2025
·
IF 13.7
Xie Jiali,Tian Jiayu,Khan Salman,Chen Feilong,Yu Jingwei,Hao Yuqiong,Xiong Hao-Ming,Zhang Feng,Zhou Qian,Zhu Guo-Yuan,Huang Ancheng C.
Glyceollins are structurally complex potent antimicrobial isoflavonoid phytoalexins produced by the crop soybean ( Glycine max ), yet their biosynthesis remains elusive, making it impossible to carry out synthetic biology-based production and engineering for further development. Here, via assembling synergistic engineering strategies, we successfully rewired the metabolic fluxes in Nicotiana benthamiana leaves for high-yield production of isoflavonoid precursor daidzein (7.04 g kg −1 dry weight (dw)), allowing for efficient screening and identification of six cytochrome P450 monooxygenases, namely glyceollin synthases, that furnish the pyrano/furano E ring and complete the 15-step biosynthetic pathways of diverse glyceollins. We establish that purified glyceollins are important for plant defense as they can effectively suppress the growth of Phytophthora sojae in vitro. Our engineered plant chassis can provide facile access to bioactive isoflavonoids, as manifested by the de novo total biosynthesis of glyceollins (for example, I, II, III and VII at up to 5.9 g kg −1 , dw) and medicarpin (0.72 g kg −1 , dw) for enhanced pathogen resistance and medicinal value.
研究领域:
植物学
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代谢工程
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合成生物学
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药用植物
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高影响因子
DRUG METABOLISM AND DISPOSITION
·
2020
·Vol. 48
·期 5
·345-352
·
IF 3.231
Xiaohua Zhao,Hong Ma,Qiusha Pan,Haiyi Wang,Xingkai Qian,Peifang Song,Liwei Zou,Mingqing Mao,Shuyue Xia,Guangbo Ge,Ling Yang
Doxophylline (DOXO) and theophylline are widely used as bronchodilators for treating asthma and chronic obstructive pulmonary disease, and DOXO has a better safety profile than theophylline. How DOXO’s metabolism and disposition affect its antiasthmatic efficacy and safety remains to be explored. In this study, the metabolites of DOXO were characterized. A total of nine metabolites of DOXO were identified in vitro using liver microsomes from human and four other animal species. Among them, six metabolites were reported for the first time. The top three metabolites were theophylline acetaldehyde (M1), theophylline-7-acetic acid (M2), and etophylline (M4). A comparative analysis of DOXO metabolism in human using liver microsomes, S9 fraction, and plasma samples demonstrated the following: 1) The metabolism of DOXO began with a cytochrome P450 (P450)–mediated, rate-limiting step at the C ring and produced M1, the most abundant metabolite in human liver microsomes. However, in human plasma, the M1 production was rather low. 2) M1 was further converted to M2 and M4, the end products of DOXO metabolism in vivo, by non-P450 dismutase in the cytosol. This dismutation process also relied on the ratio of NADP+/NADPH in the cell. These findings for the first time elucidated the metabolic sites and routes of DOXO metabolism in human.
研究领域:
药理学
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生物化学
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高影响因子
危险品化学品经营许可证（带存储）