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ENZYME AND MICROBIAL TECHNOLOGY · 2022 · IF 3.705

Specific and efficient hydrolysis of all outer glucosyls in protopanaxadiol type and protopanaxatriol type ginsenosides by a β-glucosidase from Thermoclostridium stercorarium

Cheng Zeng,Xiangqing Ji,Yu Shi,Siyu Mu,Yicheng Huang,Meiqiao Zhong,Yan Han,Cuicui Duan,Xiaolei Li,Dan Li

To develop a new method for enzymatic preparation of minor ginsenosides , T. stercorarium β-glucosidase (Tsbgl) was characterized and its activities of deglycosylation towards natural ginsenosides were examined. The substrates of 1 mmol l −1 were incubated with the enzyme of 38.3 U ml −1 at 65 ℃ and pH 5.0. The K m values of Tsbgl for ginsenosides Rb 1 , Rg 1 and p NPG were 0.37 ± 0.03, 3.26 ± 0.19, and 1.24 ± 0.03 mmol l −1 , and the V max values were 183.63 ± 7.15, 85.03 ± 4.90, and 117.66 ± 1.96 μmol mg −1 min −1 , respectively. The molar conversion of ginsenosides Rb 1 , Rb 2 , Rb 3 , Rc, Re, Rg 1 , and Rf by Tsbgl within 6 h was 100%, 50.1%, 42.7%, 92.0%, 57.3%, 67.9%, and 76.8%, respectively. The yield of aglycone protopanaxadiol was 35.5 μmol l −1 h −1 for Rb 1 , while the yields of aglycone protopanaxatriol were 64.2 and 70.4 μmol l −1 h −1 for Rg 1 and Rf. Tsbgl with good organic solvent tolerance, mild reaction conditions and broad substrate specificity , could completely remove all outer glucosyls at the C-3 and C-20 hydroxyls of protopanaxadiol-type ginsenosides, and the C-6 and C-20 hydroxyls of protopanaxatriol-type ginsenosides through various pathways, providing a specific and efficient way to produce minor ginsenosides.

研究领域: 生物化学 · 药理学 · 酶学
✓ 已验证 DOI PubMed 高影响因子
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JOURNAL OF LIQUID CHROMATOGRAPHY & RELATED TECHNOLOGIES · 2019 · IF 0.987

A metabolomic study of Asian and American Ginseng based on RRLC-QTOF/MS methods

Yang-Fang He,Han-Qing Cai,Hong-Mei Zhang,Zhi-Hui Ren,Huan Tang,Yan Li,Xiao-Wei Zhou,Wen-Long Liu,Jin Pei,Shu-Ying Liu

In order to understand the significant differences in activities and effects between Asian ginseng (ASG) and American ginseng (AMG), it is important to study the correlation between chemical structures and biological activities in the two types of herbs. However, more attention has been paid to the analysis of ginsenosides in previous reports distinguishing between ASG and AMG. There are some other bioactive compounds besides ginsenosides, however, few studies have focused on a systematic comparison of these types of compounds. Two metabolomic methods were developed in this study by qualitative data acquisition using normal phase liquid chromatography mass spectrometry (NPLC-MS) and reverse phase liquid chromatography mass spectrometry (RPLC-MS) respectively, in combination with principal component analysis (PCA). Results show that both NPLC-MS and RPLC-MS-based metabolomic methods are feasible in composition profiling, biomarker screening as well as in discrimination of ASG and AMG. 17 ginsenosides were identified as analytical markers in RPLC-MS-based metabolomic method. In comparison, using NPLC-MS-based method, 5 ginsonosides, two amino acids as well as 1 oligosaccharide were identified as analytical markers. Therefore, RPLC-MS-based metabolomic method exhibits better profiling in ginsenosides, while NPLC-MS-based metabolomic method offers the advantage that multiple active ingredients can be determined simultaneously. The two methods are both helpful in identification of biomarker as well as in discrimination of American ginsengs from Asian ginsengs.

研究领域: 植物化学 · 分析化学 · 中药学
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Food Science and Technology · 2020 · IF 1.443

Improved bioactivity and composition of Cordyceps militaris cultured with Panax ginseng

LI Li,ZUO Jin-Hui,YI Fan,YANG Yun-Li,DONG Yin-Mao,LI Qin-Yu,LI Min-Hui

Cordyceps militaris is a commonly used edible and medicinal fungus in East Asia. Preparation of the mycelium and fermentation liquid by submerged fermentation using additives such as Panax ginseng for skin whitening applications has attracted wide attention. In the present study, we screened traditional Chinese medicines as additives to the liquid fermentation medium for C. militaris and performed systematical research on the changes of bioactivity and ginsenoside levels during the fermentation process. Total polysaccharide contents, cordycepin content, and polysaccharide molecular weight in the fermentation products were measured by UV spectrophotometry, high-performance liquid chromatography-diode array detection (HPLC/DAD), and gel permeation chromatography-multi angle laser light scattering (GPC-MALLS), respectively. In addition, the ginsenosides Rb1, Re, Rd, Rg1, and pseudoginsenoside F11 in the fermentation products were quantitatively analyzed by ultra-performance liquid chromatography/tandem mass spectrometry (UPLC/MS/MS). Panax ginseng significantly increased the cordycepin and total sugar contents, and the average molecular weight of polysaccharides in the fermentation liquid and the mycelium of C. militaris. At the same time, C. militaris enhanced the production of ginsenoside pseudoginsenoside F11, which is a unique component of Panax quinquefolium. The research provides scientific evidence for the application of C. militaris and P. ginseng as raw materials for food and medicines.

CAS / MDL: 9002-10-2
研究领域: 药用真菌学 · 生物化学 · 中药学
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Journal of Ginseng Research · 2021 · IF 6.06

Evaluation of storage period of fresh ginseng for quality improvement of dried and red processed varieties

Na Zhang,Xin Huang,Yun-Long Guo,Hao Yue,Chang-Bao Chen,Shu-Ying Liu

Background Dried and red ginseng are well-known types of processed ginseng and are widely used as healthy food. The dried and red ginseng quality may vary with the storage period of raw ginseng. Therefore, herein, the effect of the storage period of fresh ginseng on processed ginseng quality was evaluated through multicomponent quantification with statistical analysis. Methods A method based on ultrahigh performance liquid chromatography coupled to triple quadrupole mass spectrometry in multiple-reaction monitoring mode (UPLC-MRM-MS) was developed for quantitation of ginsenosides and oligosaccharides in dried and red ginseng. Principal component analysis and partial least squares discriminant analysis were conducted to evaluate the dynamic distributions of ginsenosides and oligosaccharides after different storage periods. Results Eighteen PPD, PPT and OLE ginsenosides and nine reducing and nonreducing oligosaccharides were identified and quantified. With storage period extension, the ginsenoside content in the processed ginseng increased slightly in the first 2 weeks and decreased gradually in the following 9 weeks. The content of reducing oligosaccharides decreased continuously as storage time extending, while that of the nonreducing oligosaccharides increased. Chemical conversions occurred during storage, based on which potential chemical markers for the storage period evaluation of fresh ginseng were screened. Conclusion According to ginsenoside and oligosaccharide distributions, it was found that the optimal storage period was 2 weeks and that the storage period of fresh ginseng should not exceed 4 weeks at 0 °C. This study provides deep insights into the quality control of processed ginseng and comprehensive factors for storage of raw ginseng.

研究领域: 药用植物学 · 食品科学 · 中药学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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Journal of Food Measurement and Characterization · 2024 · IF 2.9

Identification and visualization of component changes during the processing of ginseng to red ginseng using UPLC-Q-Orbitrap HRMS and MALDI-MSI techniques

Wang Jin,Ren Qiao,Zhou Houqin,He Chenghao,Huang Qinwan

Red ginseng, a processed product of ginseng, has been shown to possess various pharmacological effects. To investigate the changes in ginsenosides during the processing of ginseng into red ginseng and to visualize their spatial distribution characteristics in tissues, ultra-high performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was used to detect and identify the extracts of ginseng and red ginseng. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was used to visually analyze the spatial distribution and temporal changes of prototype ginsenosides and their metabolites in ginseng. The UPLC-Q-Orbitrap HRMS results showed the identification of 42 major ginsenosides in both ginseng and red ginseng. Additionally, PCA and OPLS-DA analyses identified the processing-related markers for ginseng and red ginseng. During the processing of ginseng into red ginseng, the content of protopanaxadiol ginsenosides decreased (Rb1, Rc, Rd, etc.), while the types and content of rare ginsenosides increased (Rg3, Rg5, Rh1, etc.). The MALDI-MSI tissue distribution results demonstrated that ginsenosides in ginseng were mainly distributed in the cambium and phloem, while the processing led to the transformation of ginsenosides and an increase in distribution in the xylem and medulla. This study comprehensively reveals the chemical diversity and dynamic transformation patterns during the processing of ginseng, providing more substantial foundational information for research on red ginseng processing techniques. The transformation of these rare ginsenosides also indicates that ginseng is more beneficial for pharmacological applications such as anti-tumor, anti-allergic, and liver protection after being processed into red ginseng. Graphical abstract

CAS / MDL: 490-79-9
研究领域: 药用植物学 · 分析化学 · 中药学
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ARCHIVES OF MICROBIOLOGY · 2024 ·Vol. 206 ·期 4 ·1-10 · IF 2.3

Biotransformation approach to produce rare ginsenosides F1, compound Mc1, and Rd2 from major ginsenosides

Diao Mengxue,Chen Yanchi,Meng Lijun,Li Jianxiu,Xie Nengzhong

The stems and leaves of Panax notoginseng contain high saponins, but they are often discarded as agricultural waste. In this study, the predominant ginsenosides Rg1, Rc, and Rb2, presented in the stems and leaves of ginseng plants, were biotransformed into value-added rare ginsenosides F1, compound Mc1 (C-Mc1), and Rd2, respectively. A fungal strain YMS6 ( Penicillium sp.) was screened from the soil as a biocatalyst with high selectivity for the deglycosylation of major ginsenosides. Under the optimal fermentation conditions, the yields of F1, C-Mc1, and Rd2 were 97.95, 68.64, and 79.58%, respectively. This study provides a new microbial resource for the selective conversion of protopanaxadiol-type and protopanaxatriol-type major saponins into rare ginsenosides via the whole-cell biotransformation and offers a solution for the better utilization of P . notoginseng waste.

研究领域: 药用植物学 · 微生物学 · 天然产物化学
✓ 已验证 DOI PubMed 高影响因子
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Frontiers in Bioengineering and Biotechnology · 2024 · IF 4.3

Exploring the allelopathic autotoxicity mechanism of ginsenosides accumulation under ginseng decomposition based on integrated analysis of transcriptomics and metabolomics

Wang Rui,Zhou Tingting,Wang Yikai,Dong Jinxu,Bai Yutao,Huang Xin,Chen Changbao

Continuous cropping obstacles seriously constrained the sustainable development of the ginseng industry. The allelopathic autotoxicity of ginsenosides is the key “trigger” of continuous cropping obstacles in ginseng. During harvest, the ginseng plants could be broken and remain in the soil. The decomposition of ginseng residue in soil is one of the important release ways of ginsenosides. Therefore, the allelopathic mechanism of ginsenosides through the decomposed release pathway needs an in-depth study. To investigate this allelopathic regulation mechanism, the integrated analysis of transcriptomics and metabolomics was applied. The prototype ginsenosides in ginseng were detected converse to rare ginsenosides during decomposition. The rare ginsenosides caused more serious damage to ginseng hairy root cells and inhibited the growth of ginseng hairy roots more significantly. By high-throughput RNA sequencing gene transcriptomics study, the significantly differential expressed genes (DEGs) were obtained under prototype and rare ginsenoside interventions. These DEGs were mainly enriched in the biosynthesis of secondary metabolites and metabolic pathways, phytohormone signal transduction, and protein processing in endoplasmic reticulum pathways. Based on the functional enrichment of DEGs, the targeted metabolomics analysis based on UPLC-MS/MS determination was applied to screen endogenous differential metabolized phytohormones (DMPs). The influence of prototype and rare ginsenosides on the accumulation of endogenous phytohormones was studied. These were mainly involved in the biosynthesis of diterpenoid, zeatin, and secondary metabolites, phytohormone signal transduction, and metabolic pathways. After integrating the transcriptomics and metabolomics analysis, ginsenosides could regulate the genes in phytohormone signaling pathways to influence the accumulation of JA, ABA, and SA. The conclusion was that the prototype ginsenosides were converted into rare ginsenosides by ginseng decomposition and released into the soil, which aggravated its allelopathic autotoxicity. The allelopathic mechanism was to intervene in the response regulation of genes related to the metabolic accumulation of endogenous phytohormones in ginseng. This result provides a reference for the in-depth study of continuous cropping obstacles of ginseng.

研究领域: 植物学 · 药理学 · 分子生物学
✓ 已验证 DOI 高影响因子
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Frontiers in Pharmacology · 2022 · IF 5.988

Identification of key pharmacodynamic markers of American ginseng against heart failure based on metabolomics and zebrafish model

Dong Rong,Zhang Yougang,Chen Shanjun,Wang Huan,Hu Kaiqing,Zhao Huanxin,Tian Qingping,Zeng Kewu,Wang Songsong,Han Liwen

Background: American ginseng (Panax quinquefolium L., AG) is a traditional Chinese medicine with multiple cardiovascular protective properties. Many bioactive components have been discovered in AG over these years. However, the understanding of these key pharmacodynamic components of activity against heart failure is insufficient.Methods: A heart failure model was established using AB line wild-type zebrafish (Danio rerio) to evaluate the anti-heart failure activity of AG. Untargeted metabolomics analysis based on ultra-high performance liquid chromatography-quadrupole electrostatic field orbitrap-mass spectrometry technology (UHPLC-QE-Orbitrap-MS) was performed to screen differential components from AG samples. The potential active components were verified using the zebrafish model. Simultaneously, network pharmacology and molecular docking techniques were used to predict the possible mechanism. Finally, the key targets of six key pharmacodynamic components were verified in zebrafish using quantitative real-time-polymerase chain reaction (Q-PCR) techniques.Results: The heart failure model was successfully established in 48 h of post-fertilization (hpf) zebrafish larvae by treating with verapamil hydrochloride. The zebrafish assay showed that the anti-heart failure effects of AG varied with producing regions. The result of the herbal metabolomic analysis based on UHPLC-QE-Orbitrap-MS indicated that ginsenoside Rg3, ginsenoside Rg5, ginsenoside Rg6, malic acid, quinic acid, L-argininosuccinic acid, 3-methyl-3-butenyl-apinosyl (1→6) glucoside, pseudoginsenoside F11, and annonaine were differential components, which might be responsible for variation in efficacy. Further analysis using zebrafish models, network pharmacology, and Q-PCR techniques showed that ginsenoside Rg3, ginsenoside Rg5, ginsenoside Rg6, malic acid, quinic acid, and pseudoginsenoside F11 were the pharmacodynamic markers (P-markers) responsible for anti-heart failure.Conclusion: We have rapidly identified the P-markers against heart failure in AG using the zebrafish model and metabolomics technology. These P-markers may provide new reference standards for quality control and new drug development of AG.

研究领域: 药理学 · 心脏病学 · 代谢组学 · 斑马鱼模型
✓ 已验证 产品关联 DOI PubMed 高影响因子
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FOOD CHEMISTRY · 2025 · IF 9.8

Enhancement in the intestinal absorption of ginsenoside Re by ginseng polysaccharides and its mechanisms

Yan Zhu,Jiahui Yue,Ruyu Yan,Wenxin Xia,Tingting Li,Xueyan Fu

Ginseng, a widely used functional food and herbal medicine, contains ginsenosides as its primary active component. However, the low oral bioavailability of ginsenosides limits their therapeutic applications. Recent studies show that ginsenoside polysaccharides (GPSs) promote metabolism and biotransformation. Therefore, the objectives of this study are to investigate whether GPSs could effectively improve the oral absorption of ginsenoside components and to explore the underlying mechanisms of this effect. A GPS–ginsenoside Re complex was prepared and characterized. In vivo and in vitro experiments verified its absorption-enhancing effect. Results showed that the GPS–ginsenoside Re complex enhanced absorption through hydrogen bonding and protected ginsenoside Re from degradation. Mechanistic studies revealed that the GPS–ginsenoside Re complex promoted the absorption of ginsenoside Re by regulating the expression of four efflux proteins and two tight junction proteins. Therefore, GPSs can enhance the bioavailability and efficacy of ginsenosides, particularly for liver-related diseases.

CAS / MDL: 108-95-2 7664-93-9
研究领域: 药理学 · 中药学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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DRUG DEVELOPMENT AND INDUSTRIAL PHARMACY · 2020 · IF 2.365

Panax quinquefolium saponin liposomes prepared by passive drug loading for improving intestinal absorption

Bing Zhang,Wenli Pan,Yaxin Deng,Haibing He,Jingxin Gou,Yanjiao Wang,Yu Zhang,Tian Yin,Dongchun Liu,Xing Tang

Panax quinquefolium saponin (PQS) composed of 45% pseudo-ginsenoside F11 (PF11), is a natural mixture of sterol compounds obtained from the American ginseng plant, having numerous promising benefits for health. However, low solubility and permeability limit the development of PQS as a therapeutic agent for oral administration. In this study, PQS liposomes (PQS-Lips) were prepared by thin layer hydration, an in situ single-pass intestinal perfusion (SPIP) model was used to verify the improvement of membrane permeability of PQS-Lips. PQS-Lips had a high encapsulation efficiency (EE) of 65%∼70%, a particle size about 100.0 nm, and a zeta potential of −60 mV with regular spherical surface. FTIR and DSC showed the PQS in liposomes were amorphous, indicating that hydrogen bonds formed between one or several hydroxyl groups in PQS and C-O group at the phospholipid polar terminal. In addition, PQS-Lips showed sustained release in vitro than PQS at pH 1.2 and pH 6.8, and PQS-Lips had good stability in simulated gastric and intestinal fluid. Then, the absorption rate (Ka) and effective permeability coefficient (Peff) of PQS-Lips in the whole small intestine were significantly higher than those in PQS solution (PQS-Sol), which proved that the PQS-Lips could significantly increase the membrane permeability of PQS and promote its absorption in the small intestine. From the experimental results, it could be known that liposome technology could effectively improve the absorption of PQS in the small intestine.

CAS / MDL: 9002-07-7
研究领域: 药剂学 · 生物药剂学 · 中药学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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APPLIED MICROBIOLOGY AND BIOTECHNOLOGY · 2024 ·Vol. 108 ·期 1 ·1-13 · IF 3.9

Novel glycosidase from Paenibacillus lactis 154 hydrolyzing the 28-O-β-d-glucopyranosyl ester bond of oleanane-type saponins

Wu Zongzhan,Dou Wenyu,Yang Xiaolin,Niu Tengfei,Han Zhuzhen,Yang Li,Wang Rufeng,Wang Zhengtao

Abstract Oleanane-type ginsenosides are a class of compounds with remarkable pharmacological activities. However, the lack of effective preparation methods for specific rare ginsenosides has hindered the exploration of their pharmacological properties. In this study, a novel glycoside hydrolase Pl GH3 was cloned from Paenibacillus lactis 154 and heterologous expressed in Escherichia coli . Sequence analysis revealed that Pl GH3 consists of 749 amino acids with a molecular weight of 89.5 kDa, exhibiting the characteristic features of the glycoside hydrolase 3 family. The enzymatic characterization results of Pl GH3 showed that the optimal reaction pH and temperature was 8 and 50 °C by using p -nitrophenyl- β - d -glucopyranoside as a substrate, respectively. The K m and k cat values towards ginsenoside Ro were 79.59 ± 3.42 µM and 18.52 s −1 , respectively. Pl GH3 exhibits a highly specific activity on hydrolyzing the 28- O - β - d -glucopyranosyl ester bond of oleanane-type saponins. The mechanism of hydrolysis specificity was then presumably elucidated through molecular docking. Eventually, four kinds of rare oleanane-type ginsenosides (calenduloside E, pseudoginsenoside RP1, zingibroside R1, and tarasaponin VI) were successfully prepared by biotransforming total saponins extracted from Panax japonicus . This study contributes to understanding the mechanism of enzymatic hydrolysis of the GH3 family and provides a practical route for the preparation of rare oleanane-type ginsenosides through biotransformation. Key points • The glucose at C-28 in oleanane-type saponins can be directionally hydrolyzed. • Mechanisms to interpret PlGH3 substrate specificity by molecular docking. • Case of preparation of low-sugar alternative saponins by directed hydrolysis.

研究领域: 生物化学 · 药理学 · 微生物学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY · 2024 · IF 5.7

Desorption Electrospray Ionization-Mass Spectrometry Imaging-Based Spatial Metabolomics for Visualizing and Comparing Ginsenosides and Lipids among Multiple Parts and Positions of the Panax ginseng Root

Hongda Wang,Lili Hong,Feifei Yang,Yuying Zhao,Qi Jing,Wei Wang,Min Zhang,Yang Yang,Qinhua Chen,Ying Hu,Yadan Zou,Xue Li,Wenzhi Yang

Ginsenosides and lipids are both bioactive ingredients for ginseng. Targeting these two categories of components, this study was designed to develop desorption electrospray ionization-mass spectrometry imaging approaches and spatial metabolomics strategies, achieving the visualization and differentiation among different parts of Panax ginseng root (e.g., rhizome, main root, lateral root, fibrous root, and adventitious root). Potential chemical markers were identified by searching an in-house ginsenoside library and online Lipid Maps database, together with high-resolution MS2 data analysis. Six ginsenosides and 11 lipids were diagnostic to differentiate five different parts of the P. ginseng root. Additionally, three ginsenosides and 20 lipids were identified as differential markers among the six positions of the main root of P. ginseng. High-abundance malonyl- and oleanolic acid-ginsenosides were observed in the rhizome. These results assist in understanding the accumulation of bioactive molecules all through the root of P. ginseng, which can benefit its quality control and rational use.

CAS / MDL: 631-61-8
研究领域: 植物化学 · 分析化学 · 中药学
✓ 已验证 产品关联 DOI PubMed 高影响因子
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