PLANT PHYSIOLOGY
·
2023
·
IF 7.4
Liao Zhenkun,Liu Xiaojuan,Zheng Juan,Zhao Chenning,Wang Dengliang,Xu Yang,Sun Chongde
Polymethoxylated flavones (PMFs) have received extensive attention due to their abundant bioactivities. Citrus peels specifically accumulate abundant PMFs, and methylation modification is a key step in PMF biosynthesis; however, the function of reported O-methyltransferase (OMT) in citrus is insufficient to elucidate the complete methylation process of PMFs. In this study, we analyzed the accumulation pattern of PMFs in the flavedo of the sweet orange (Citrus sinensis) cultivar “Bingtangcheng” at different developmental stages. We found that accumulation of PMFs was completed at the early stage of fruit development (60-d after flowering). Furthermore, we characterized a true caffeoyl-CoA O-methyltransferase (named CsCCoAOMT1) from C. sinensis. Functional analysis in vitro showed that CsCCoAOMT1 preferred flavonoids to caffeoyl-CoA and esculetin. This enzyme efficiently methylated the 6-, 7- 8-, and 3′-OH of a wide array of flavonoids with vicinal hydroxyl groups with a strong preference for quercetin (flavonol) and flavones. The transient overexpression and virus-induced gene silencing experiments verified that CsCCoAOMT1 could promote the accumulation of PMFs in citrus. These results reveal the function of true CCoAOMTs and indicate that CsCCoAOMT1 is a highly efficient multifunctional O-methyltransferase involved in the biosynthesis of PMFs in citrus.
研究领域:
植物生物化学
·
分子生物学
·
植物生理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MOLECULES
·
2020
·Vol. 25
·期 4
·858
·
IF 3.267
Xiaojuan Liu,Yue Wang,Yezhi Chen,Shuting Xu,Qin Gong,Chenning Zhao,Jinping Cao,Chongde Sun
O-methylation of flavonoids is an important modification reaction that occurs in plants.O-methylation contributes to the structural diversity of flavonoids, which have several biological and pharmacological functions. In this study, anO-methyltransferase gene (CrOMT2) was isolated from the fruit peel ofCitrus reticulata, which encoding a multifunctionalO-methyltransferase and could effectively catalyze the methylation of 3’-, 5’-, and 7-OH of flavonoids with vicinal hydroxyl substitutions. Substrate preference assays indicated that this recombinant enzyme favored polymethoxylated flavones (PMF)-type substrates in vitro, thereby providing biochemical evidence for the potential role of the enzyme in plants. Additionally, the cytotoxicity of the methylated products from the enzymatic catalytic reaction was evaluated in vitro using human gastric cell lines SGC-7901 and BGC-823. The results showed that the in vitro cytotoxicity of the flavonoids with the unsaturated C2-C3 bond was increased after being methylated at position 3’. These combined results provide biochemical insight regarding CrOMT2 in vitro and indicate the in vitro cytotoxicity of the products methylated by its catalytic reaction.Keywords:citrus;flavonoids;O-methyltransferase;bioactivity;cytotoxicity
研究领域:
植物生物化学
·
药理学
·
分子生物学
·
细胞生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2025
·
IF 8
Zhanming Tang,Yanyan Miao,Huimin Lu,Chao Chen,Shiming Li,Wensheng Zhang
Citri Reticulatae Pericarpium (Xinhui Chenpi) , a representative product with a protected geographical indication from Xinhui District, Guangdong Province, China, is a dual-use medicinal and edible material and a traditional medicinal resource whose quality is traditionally believed to improve with prolonged aging. Increasing evidence indicates that the succession of surface microbial communities and their metabolic activities play pivotal roles in these transformations. This study systematically investigated Xinhui Chenpi aged 1–10 years to elucidate the dynamics of dominant fungal genera and experimentally verify their involvement in flavonoid transformation. Ultra-performance liquid chromatography (UPLC), third-generation full-length ITS sequencing (PacBio) , and strain isolation with culture assays were employed to characterize flavonoid profiles, fungal community succession, and the transformation capabilities of key isolates. Flavonoids exhibited a “degradation to resynthesis” trajectory, with polymethoxyflavones (PMFs) steadily accumulating throughout the aging process. Late-stage samples were dominated by the xerophilic genus Xeromyces (Aspergillaceae), which showed strong positive correlations with PMF enrichment, whereas correlations with Penicillium progressively declined over time. Importantly, Aspergillus tubingensis was isolated and confirmed to convert nobiletin into 3′-demethylnobiletin, providing direct mechanistic evidence for fungal-driven PMF structural modification. Community profiling further revealed that multiple Aspergillaceae genera ( Aspergillus , Penicillium , Xeromyces ) may participate in flavonoid transformation. These findings elucidate the microbial mechanisms underlying quality enhancement in aged Chenpi and provide a scientific basis for microbial-assisted optimization of modern aging processes, thereby offering theoretical support for the traditional principle that “aged Chenpi becomes more efficacious.”
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Food Measurement and Characterization
·
2021
·Vol. 15
·期 5
·3873-3883
·
IF 2.431
Qian Zheng-ming,Huang Qi,Feng Xiao-long,Li De-qiang
Given the safety of synthetic antioxidants, people are increasingly inclined to develop and use natural antioxidants. However, rapid screening and identification of active ingredients from the complex systems of natural products is one of the major challenges for the modernization of traditional Chinese medicine. In this study, online extraction-high performance liquid chromatography-diode array detection-quadrupole time-of-flight mass spectrometry coupled with ABTS antioxidant assay (OLE-HPLC-DAD-QTOF-MS/MS-ABTS), in which sample preparation, separation, detection and antioxidant assay were integrated in a single step, was used to rapidly identify antioxidant components from the different parts of Citrus reticulata , including its peels, leaves, vascular bundies and seeds. After method validation (system adaptability, extraction yield and repeatability), the four samples of different botanical parts were analyzed. As a result, among 49 found chromatographic peaks, 33 peaks displayed varying degrees of antioxidant activity and 39 peaks were tentatively or unambiguously identified based on their retention time, UV spectrum and mass spectrometry information and the comparison with reference substance and data in literature, including 10 phenolic acids, 9 polymethoxy flavonoids, 7 flavonoid- O -glycosides and 13 other ingredients. It is concluded that the established method is simple, fast, and reliable, which not only saves the materials and solvents as well as time, but also reduces the risk of degradation of chemical components during the extraction process, suitable for rapid screening and identification of antioxidant components of complex mixture.
研究领域:
生药学
·
分析化学
·
天然产物
·
中药学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF EXPERIMENTAL BOTANY
·
2020
·Vol. 71
·期 10
·3066-3079
·
IF 5.908
Liu Xiaojuan,Zhao Chenning,Gong Qin,Wang Yue,Cao Jinping,Li Xian,Grierson Donald,Sun Chongde
Polymethoxylated flavones (PMFs), which accumulate exclusively in fruit peel of citrus, play important physiological and pharmacological roles but the genetic basis for the methylation of flavonoids has not been fully elucidated in citrus. Here we characterize a caffeoyl-CoA O-methyltransferase-like enzyme, designated CrOMT1. The expression pattern of CrOMT1 was highly correlated with the concentration of the three major PMFs in two different citrus fruit tissues during fruit maturation. Exposure of fruit to UV-B radiation sharply increased the level of CrOMT1 transcripts and also led to the accumulation of three PMFs. The potential role of CrOMT1 was studied by testing the catalytic activity of recombinant CrOMT1 with numerous possible substrates in vitro. The enzyme could most efficiently methylate flavones with neighboring hydroxy moieties, with high catalytic efficiencies found with 6-OH- and 8-OH-containing compounds, preferences that correspond precisely with the essential methylation sites involved in the synthesis of the three naturally occurring PMFs in Citrus reticulata. This indicates that CrOMT1 is capable of in vitro methylation reactions required to synthesize PMFs in vivo. Furthermore, transient overexpression of CrOMT1 increased levels of the three major PMFs in fruit, indicating that CrOMT1 is likely to play an essential role in the biosynthesis of PMFs in citrus.
研究领域:
植物生物学
·
生物化学
·
分子遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Food Bioscience
·
2025
·
IF 5.9
Hongyang Chen,Zimao Ye,Yanchi Liu,Zhiqin Zhou
An integrated physiological and non-targeted metabolomics approach was used to construct a eugenol-based polymethoxyflavones nanoemulsion delivery system and examine its increased antibacterial activity on E. coli . Compared with pure eugenol nanoemulsion (EG, MIC: 640 μg/mL), the polymethoxyflavones-eugenol nanoemulsion (EGP) exhibited significantly enhanced antimicrobial activity (MIC: 320 μg/mL). EGP treatment disrupted the integrity of E. coli cell membranes and walls, increased cell permeability. Non-targeted metabolomics analysis (compared with CK) revealed 685 upregulated and 642 downregulated differentially accumulating metabolites, lysine and glutamine are reduced in E. coli , and glutaric acid, aminoadipic acid, and D-lactic acid are increased. Pathway enrichment analysis showed that amino acid metabolism and energy metabolism were significantly disturbed by TCA cycle disruption. EGP exhibits superior antimicrobial efficacy against E. coli through a multi-targeted mechanism involving cell membrane disruption and metabolic pathway interference. This study provides valuable insights into the development of natural antimicrobial nanoemulsion as potential alternatives to synthetic preservatives for food safety applications.
研究领域:
食品科学
·
微生物学
·
纳米技术
·
药理学
·
代谢组学
✓ 已验证
产品关联
DOI
高影响因子
Fungal Biology
·
2022
·
IF 2.91
Long Guo,Xiaoxue Mao,Yi Li,Zhiqin Zhou
Citrus is susceptible to Penicillium digitatum ( P. digitatum ) infection in post-harvest storage, resulting in enormous economic losses. This study aimed to investigate the antifungal activity and potential mechanism of the combination of Polymethoxylated flavones (PMFs) and citral (two natural antifungal components derived from citrus) against P. digitatum in vitro and citrus fruit. The results show that PMFs can enhance the antifungal activity of citral nanoemulsion, and PMFs-loaded citral nanoemulsion (PCT) has significant antifungal activity in a concentration-dependent manner. PCT can evidently inhibit spore germination and mycelial growth in vitro , and effectively control the growth of green mold on postharvest citrus fruit. Furthermore, PCT treatment resulted in the alteration of mycelia morphology, accumulation of reactive oxygen species, and membrane lipid peroxidation. These changes can disrupt the normal structure and function of the cell membrane, as evidenced by the reduction of total lipid and ergosterol content in the mycelia and the stronger red fluorescence of the cells emitted after PI staining. Based on the above results, we infer that PCT has a strong inhibitory effect on P. digitatum , and its potential mechanism is related to the destruction of the cell membrane. Therefore, PCT can be considered as a botanical fungicide for citrus preservation.
研究领域:
植物科学
·
食品科学
·
微生物学
·
植物病理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
·
2024
·
IF 8.5
Hanliang Li,Lianzhu Lin,Mouming Zhao
This study aimed at recycling flavonoids with glycolipid lowering activity from Chenpi processing residue produced by essential oil extraction. The crude Chenpi flavonoids obtained by ethanol reflux mainly contained flavonone (hesperidin) and polymethoxyflavonoids (nobiletin, tangeretin, sinensetin, and isosinensetin), which complementally exerted strong glycolipid lowering activity as revealed by digestive enzyme inhibitory activity, molecular docking and network pharmacology. The reducing sugar and water-soluble impurities were efficiently removed by yeast fermentation and aqueous two-phase system (ATPS) extraction, respectively. The optimal combination use of yeast (2 % w / v , CN36) and ATPS formed by 22 % ( w /w) (NH 4 ) 2 SO 4 and 25 % (w/w) ethanol improved flavonoid purity to 60.25 %, enriched flavonone and polymethoxyflavonoids by 6.69-fold, and enhanced digestive enzyme inhibitory activity, which was superior to the use of individual yeast or ATPS, unoptimized combination, macroporous resin (XAD-16), and ethyl acetate. This study was the first to develop a recyclable biophysical method for simultaneous enrichment of flavonone and polymethoxyflavonoids.
研究领域:
药学
·
生物技术
·
食品科学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2022
·
IF 5.895
Xiaoqi Wang,Dongli Li,Yong Cao,Chi-Tang Ho,Qingrong Huang
5-Demethylated polymethoxyflavones (5-OH PMFs) are the most unique monodemethylated PMFs with relatively low polarities and are proved to possess better anticancer and anti-inflammatory effects than their respective permethoxylated ones. However, their detailed in vivo metabolic fates have not been fully studied. 5-Demethylsinensetin (5-OH Sin), being one of the 5-demethylated citrus PMFs, was used in the present research to investigate its biotransformation in pharmacokinetics and excretion in rats. The results showed that 5-OH Sin was mostly accumulated in the large intestine, indicating its poor absorption in the small intestine. In addition, 5,3′-didemethylsinensetin and 5,4′-didemethylsinensetin were identified as two dominated metabolites of 5-OH Sin, and the C-3′ position of 5-OH Sin was more facile to be demethylated in systemic circulation. Moreover, other than demethylation reactions, the methylation transformation of 5-OH Sin and its metabolites were also observed and quantified, suggesting that the bidirectional biotransformation between 5-OH Sin and its parent compound, Sin, occurred under in vivo conditions.
研究领域:
药理学
·
生物化学
·
药学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Journal of Fungi
·
2022
·Vol. 8
·期 4
·388
·
IF 5.724
Long Guo,Yi Li,Xiaoxue Mao,Rui Tao,Boyun Tao,Zhiqin Zhou
A major citrus postharvest pathogen,Penicilliumitalicum(P. italicum), causes substantial economic losses in citrus. In this study, a citral nanoemulsion containing polymethoxylated flavonoids (PMFs), the antimicrobial compounds from citrus, was prepared. The antifungal activity and potential antifungal mechanisms of the nanoemulsion againstP. italicumwere evaluated. The results showed that the growth ofP. italicumwas effectively inhibited by the nanoemulsion, with a minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of 62.5 and 250 mg L−1, respectively. The nanoemulsion significantly inhibited spore germination and mycelial growth, and it altered the morphology ofP. italicum. In addition, the permeability of the cell membrane increased with increasing nanoemulsion concentrations, as evidenced by a rapid rise in extracellular electric conductivity and stronger red fluorescence from mycelia (propidium iodide staining). Compared with the control, the nanoemulsion treatment induced a decrease in total lipid and ergosterol contents inP. italicumcells by 64.61% and 60.58%, respectively, demonstrating that membrane integrity had been disrupted. The results indicated that the PMFs-loaded nanoemulsion exerted antifungal activity againstP. italicumby disrupting cell membrane integrity and permeability; such a nanoemulsion may be used as a potential fungicide substitute for preservation in citrus fruits.Keywords:antifungal mechanism;citrus blue mold;membrane integrity;membrane permeability;P. italicum
研究领域:
植物病理学
·
食品科学
·
药理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF FOOD AND DRUG ANALYSIS
·
2022
·
IF 6.157
Ping-Ting Xiao,Yu-Jia Kuang,Shi-Yu Liu,Zhi-Shen Xie,Jin-Hua Hao,E-Hu Liu
Since the combinatorial components responsible for the antihyperlipidemic activity of Citrus reticulata ‘Chachi’ (CRC) peels remains unclear, we herein developed a bioactive equivalence oriented feedback screening method to discover the bioactive equivalent combinatorial components (BECCs) from CRC peels. Using palmitic acid (PA)-stimulated hepatocyte model, a combination of 5 polymethoxyflavones (PMFs) including tangeretin, sinensetin, nobiletin, 5,7,8,4′-tetramethoxyflavone and 3,5,6,7,8,3′,4′-heptamethoxyflavone was identified to be responsible for the antihyperlipidemic effect of CRC peels. Via evaluation of combination effect by combination index (CI), these 5 PMFs were found to take effect via a synergistic mode. Our data indicated that the antihyperlipidemic mechanism of PMF combination was associated with the inhibition of fatty acid and cholesterol synthesis, and inflammation. Also, the PMF combination exhibited robust antihyperlipidemic effects in HFD-fed rats in vivo . Our study offers evidence-based data to uncover the pharmacological effect of CRC peels.
研究领域:
药理学
·
生物化学
·
营养学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2021
·
IF 4.952
Lingrong Wen,Mingyang He,Chunxiao Yin,Yueming Jiang,Donghui Luo,Bao Yang
Sour orange ( Citrus aurantium ) is a popular fruit with health benefits. Phenolics are the dominant bioactive compounds in this fruit. However, the full phenolics composition remains unclear. In this work, UHPLC-MS/MS was used to qualify and quantify the phenolics in sour orange fruits of cv. Morocco (MSO) and United (USO). Fifty-eight phenolics, including 26 flavonoid glycosides , 22 polymethoxylated flavonoids, 8 coumarins and 2 phenolic acids , were identified. Four limonoids were characterized. Flavonoid glycosides were the leading phenolics in MSO, while polymethoxylated flavonoids (including nobiletin, tangeritin and sinensetin) were dominant in USO. The phenolic extracts showed good antioxidant activities . USO extract had a significantly ( p < 0.05) stronger neuroprotective activity than MSO extract. Auraptene, nobiletin and tangeritin were highly responsible for the neuroprotective activity. The results were helpful for the development of functional foods with sour orange.
研究领域:
食品科学
·
药理学
·
分析化学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）