Biotransformation of ginsenosides Re and Rg1 into ginsenosides Rg2 and Rh1 by recombinant β-glucosidase

Ginsenosides Re and Rg1 were transformed by recombinant β-glucosidase (Bgp1) to ginsenosides Rg2 and Rh1, respectively. The bgp1 gene consists of 2,496?bp encoding 831 amino acids which have homology to the glycosyl hydrolase families 3 protein domain. Using 0.1?mg enzyme ml(-1) in 20?mM sodium phosphate buffer at 37°C and pH 7.0, the glucose moiety attached to the C-20 position of ginsenosides Re and Rg1, was removed: 1?mg ginsenoside Re ml(-1) was transformed into 0.83?mg Rg2?ml(-1) (100% molar conversion) after 2.5?h and 1?mg ginsenoside Rg1?ml(-1) was transformed into 0.6?mg ginsenoside Rh1?ml(-1) (78% molar conversion) in 15?min. Using Bgp1 enzyme, almost all initial ginsenosides Re and Rg1 were converted completely to ginsenosides Rg2 and Rh1. This is the first report of the conversion of ginsenoside Re to ginsenoside Rg2 and ginsenoside Rg1 to ginsenoside Rh1 using the recombinant β-glucosidase.     

Bioconversion of major ginsenosides Rg(1) to minor ginsenoside F(1) using novel recombinant ginsenoside hydrolyzing glycosidase cloned from Sanguibacter keddieii and enzyme characterization

This study focuses on the cloning, expression, and characterization of recombinant ginsenoside hydrolyzing glycosidase from Sanguibacter keddieii in order to biotransform ginsenosides efficiently. The gene, termed bglSk, consists of 1857bp and revealed significant homology to that of glycoside hydrolase family 3. The enzyme was over-expressed in Escherichia coli BL21 (DE3) using a GST-fused pGEX 4T-1 vector system. The over-expressed recombinant enzymes could convert six major ginsenosides Rb(1), Rb(2), Rc, Rd, Re and Rg(1) into more pharmacologically active rare ginsenosides such as C-Y, C-Mc, C-K, Rg(2)(S), and F(1). Especially, BglSk could completely convert the Rg(1) into F(1). The GST-fused BglSk was purified with GST·bind agarose resin and then characterized. The kinetic parameters for β-glucosidase had apparent K(m) values of 0.456±0.009 and 0.167±0.003mM and V(max) values of 30.2±0.7 and 4.1±0.1μmolmin(-1)mg of protein(-1) against p-nitrophenyl-β-d-glucopyranoside and Rb(1), respectively.     

Simple and efficient preparation of ginsenoside (S)-Rg2 from ginsenoside Re by biotransformation with Cellulosimicrobium sp. 21

A novel ginsenoside-hydrolyzing strain was isolated from ginseng-cultivation soil in Changbai Mountain (China). The strain was identified as Cellulosimicrobium sp. 21 by 16S rDNA sequencing. Using the β-glucosidases secreted from Cellulosimicrobium sp. 21, protopanaxatriol-type ginsenoside Re was converted to the highly active neuroprotective molecule (S)-Rg2 by removal of the C-20-glucopyranosyl residue. The α-L-rhamnopyranosyl-(1→2)-β-D-glucopyranose at the C-6 position of Rg2 was not further attacked by Cellulosimicrobium sp. 21, so the transformation shows high specificity. To simplify the transformation and product-preparation process, a simple and efficient transformation system was developed in a phosphate buffer system instead of organic media. The optimum conditions for transforming ginsenoside Re into Rg2 by Cellulosimicrobium sp. 21 were determined through single-factor experiments and response surface methodology. Under the optimized conditions: transformation buffer, 50 mM phosphate buffer, at pH: 7.00; temperature: 27.6°C; substrate concentration: 0.50 mg/ml; biotransformation period: 12 h; the biotransformation efficiency reached 89.8% (molar ratio) in 2-L reaction system. This simple biotransformation with high specificity and efficiency has potential for use in Rg2 preparation in the pharmaceutical industry.     

Hydrolysis of the outer β-(1,2)-d-glucose linkage at the C-3 position of ginsenosides by a commercial β-galactosidase and its use in the production of minor ginsenosides

Commercial β-galactosidase from Aspergillus oryzae (SUMILACT L^TM) was used for the bioconversion of the ginsenosides Rb1, Rb2, Rc, Rd, and Rg3 to gypenoside-XVII, compound-O, compound-MC1, F2, and Rh2, respectively. The optimal conditions were pH 4.5, 50?°C, 60?U·mL^?1 enzyme, and 8.0?mM substrate. Interestingly, the enzyme hydrolyzed only the outer β-(1,2)-d-glucose linkage at the C-3 position of ginsenosides. Under optimum conditions, the enzyme completely converted Rb1, Rb2, Rc, Rd, and Rg3 to gypenoside-XVII, compound-O, compound-MC1, F2, and Rh2, respectively, with the highest productivity.     

Transformation of Ginsenosides Rb1 and Re from <Emphasis Type="Italic">Panax ginseng</Emphasis> by Food Microorganisms

Ginsenosides Rb1 and Re, respectively belonging to the major protopanaxadiol and protopanaxatriol ginsenosides, were transformed using cell-free extracts from food microorganisms. Rb1 was transformed into compound K via Rd and F2 by Bifidobacterium sp. Int57, Bif. sp. SJ32, Aspergillus niger and A.␣usamii. Lactobacillus delbrueckii, and Leuconostoc paramesenteroides transformed Rb1 into Rh2 via Rd and F2. Bifidobacterium sp. SH5 transformed Rb1 into F2 via Rd. Re was transformed into Rh1 via Rg2 by Bif. sp. Int57 and Bif. sp. SJ32. A. niger transformed Re into Rh1 via Rg1. A. usamii transformed Re into Rg2. Transformation of Rb1 proceeded at a higher rate and needed less amount of enzymes than that of Re. Taken together, these processes would allow a specific bioconversion process possible to obtain specific ginsenosides using an appropriate combination of ginsenoside substrates and specific microbial enzymes.     

Neuroprotective effect of individual ginsenosides on astrocytes primary culture

Most of the known pharmacological effects of Panax ginseng on the central nervous system are due to its major components - ginsenosides. Although the antioxidant ability of ginseng root has already been established, this activity has never been evaluated for isolated ginsenosides on astrocytes. The activity of protopanaxadiols Rb(1), Rb(2), Rc and Rd, and protopanaxatriols Re and Rg(1) was evaluated in vitro on astrocytes primary culture by means of an oxidative stress model with H(2)O(2). The viability of astrocytes was determined by the MTT reduction assay and by the LDH release into the incubation medium. The effects on the antioxidant enzymes catalase, superoxide dismutase (SOD), glutathione peroxidases (GPx) and glutathione reductase (GR) and on the intracellular reactive oxygen species (ROS) formation were also investigated. Exposure of astrocytes to H(2)O(2) decreased cell viability as well as the antioxidant enzymes activity and increased ROS formation. Oxidative stress produced significant cell death that was reduced by previous treatment with the tested ginsenosides. Ginsenosides Rb(1), Rb(2), Re and Rg(1) were effective in reducing astrocytic death, while Rb(1), Rb(2), Rd, Re and Rg(1) decreased ROS formation, ginsenoside Re being the most active. Ginsenosides from P. ginseng induce neuroprotection mainly through activation of antioxidant enzymes.     

Application of microwave-irradiation technique in deglycosylation of ginsenosides for improving apoptosis induction in human melanoma SK-MEL-2 cells

To increase the contents of medicinally effective ginsenosides, we used high-temperature and high-pressure thermal processing of ginseng by exposing it to microwave irradiation. To determine the anti-melanoma effect, the malignant melanoma SK-MEL-2 cell line was treated with an extract of microwave-irradiated ginseng. Microwave irradiation caused changes in the ginsenoside contents: the amounts of ginsenosides Rg1, Re, Rb1, Rb2, Rc, and Rd were disappeared, while those of less polar ginsenosides, such as Rg3, Rg5, and Rk1, were increased. In particular, the contents of Rk1 and Rg5 markedly increased. Melanoma cells treated with the microwave-irradiated ginseng extract showed markedly increased cell death. The results indicate that the microwave-irradiated ginseng extract induced melanoma cell death via the apoptotic pathway and that the cytotoxic effect of the microwave-irradiated ginseng extract is attributable to the increased contents of specific ginsenosides.     

Can ginsenosides protect human erythrocytes against free-radical-induced hemolysis?

Many studies have focused on the free-radical-initiated peroxidation of membrane lipid, which is associated with a variety of pathological events. Panax ginseng is used in traditional Chinese medicine to enhance stamina and capacity to deal with fatigue and physical stress. Many reports have been devoted to the effects of ginsenosides, the major active components in P. ginseng, on the lipid metabolism, immune function and cardiovascular system. The results, however, are usually contradictory since the usage of mixture of ginsenosides cannot identify the function of every individual ginsenosides on the experimental system. On the other hand, every individual ginsenosides is not compared under the same experimental condition. These facts motivate us to evaluate the antioxidant effect of various individual ginsenosides on the experimental system of free-radical-initiated peroxidation: the hemolysis of human erythrocyte induced thermally by water-soluble initiator, 2,2′-azobis(2-amidinopropane hydrochloride) (AAPH). The inhibitory concentration of 50% inhibition (IC₅₀) of AAPH-induced hemolysis of the erythrocyte has been studied firstly and found that the order of IC₅₀ is Rb3∼Rb1≪Rg2<Re<Rg1∼Rc<Rh1<R1. Rb1, Rc and Rg2, as antioxidants, can prolong the lag time of hemolysis. Contrarily, Rg3, Rd and Rh1, together with high concentration of Rb3, Rg1 and Rh2, function as prooxidants to accelerate AAPH-induced hemolysis. The addition of Re does not influence the lag time of hemolysis. The R1 with the concentration ranging from 10 to 20 μM decreases the lag time of hemolysis. These results suggest that there is a mutual interaction that existed in the molecule of ginsenosides since the difference of the structure of ginsenosides is only due to the connective position and type of sugar moieties to the ring of a triterpene dammarane. Moreover, the synergistic antioxidative properties of various individual ginsenosides with α-tocopherol (TOH) are also discussed, and it was found that the order of synergistic antioxidative properties with TOH is Rb1>Rc>Re>Rh1>R1>Rg2>Rb3. Rg3, Rd and Rh2, however, act as synergistic prooxidants in the above experimental system. Rg1 does not show any synergistic antioxidative property. Although the antioxidative and prooxidative mechanism of various ginsenosides with or without TOH in AAPH-induced hemolysis of human erythrocytes will be further studied in detail, this information may be useful in the clinical usage of ginsenosides.     

黑根霉对人参皂苷Re的生物转化

利用菌种黑根霉Rhizopus sp.对人参皂苷Re进行生物转化,并对人参皂苷Re及其发酵产物进行HPLC系统分析比较,经液相色谱-质谱分析得出人参皂苷Re转化率为92.16%,并制备出人参皂苷Re发酵产物中峰值升高的成分,转化后的人参皂苷发酵产物中化合物1确定为人参皂苷Rg2,化合物2为Rg2的同分异构体,得率为10.13%;化合物3和化合物4确定为人参皂苷Rg5/Rk1,得率为29.23%。从结果初步推测得出人参皂苷Re被黑根霉转化为人参皂苷Rg2的机理,人参皂苷Re转化成人参皂苷Rg5/Rk1的机理还有待于进一步研究。     

Can ginsenosides protect human erythrocytes against free-radical-induced hemolysis?

Many studies have focused on the free-radical-initiated peroxidation of membrane lipid, which is associated with a variety of pathological events. Panax ginseng is used in traditional Chinese medicine to enhance stamina and capacity to deal with fatigue and physical stress. Many reports have been devoted to the effects of ginsenosides, the major active components in P. ginseng, on the lipid metabolism, immune function and cardiovascular system. The results, however, are usually contradictory since the usage of mixture of ginsenosides cannot identify the function of every individual ginsenosides on the experimental system. On the other hand, every individual ginsenosides is not compared under the same experimental condition. These facts motivate us to evaluate the antioxidant effect of various individual ginsenosides on the experimental system of free-radical-initiated peroxidation: the hemolysis of human erythrocyte induced thermally by water-soluble initiator, 2,2'-azobis(2-amidinopropane hydrochloride) (AAPH). The inhibitory concentration of 50% inhibition (IC(50)) of AAPH-induced hemolysis of the erythrocyte has been studied firstly and found that the order of IC(50) is Rb3 - Rb1Rc>Re>Rh1>R1>Rg2>Rb3. Rg3, Rd and Rh2, however, act as synergistic prooxidants in the above experimental system. Rg1 does not show any synergistic antioxidative property. Although the antioxidative and prooxidative mechanism of various ginsenosides with or without TOH in AAPH-induced hemolysis of human erythrocytes will be further studied in detail, this information may be useful in the clinical usage of ginsenosides.     

人参生殖器官皂苷含量动态变化

为了明确从现蕾、开花到结实过程中的人参生殖器官中各单体皂苷含量的动态变化,应用HPLC法测定了人工栽培的五年生人参不同时期生殖器官中的人参单体皂苷Rb1、Rb2、Rb3、Rc、Rd、Re、Rg1和Rg3的含量。结果显示:从现蕾到果实成熟的过程中,人参单体皂苷Rb1、Rb2、Rb3、Rc、Rd、Re、Rg1和Rg3的含量的平均值分别为0.643%,0.189%,1.026%,1.014%,1.941%,8.381%,0.724%和0.041mg.g-1。从现蕾到果实成熟的过程中,人参单体皂苷Rb1含量的最高值在7月16日,单体皂苷Rb3、Rc、Rd和Rg1含量的最高值在7月11日,单体皂苷Rb2和Rg2含量的最高值在8月7日。     

Identification and characterization of a novel bacterial β-glucosidase that is highly specific for the β-1,2-glucosidic linkage of sesaminol triglucoside

A gene (PSTG2) coding for a novel β-glucosidase belonging to glycoside hydrolase family 3 was identified in the vicinity of the previously identified β-glucosidase gene [sesaminol triglucoside (STG)-hydrolyzing β-glucosidase, PSTG1] in the genome of Paenibacillus sp. strain KB0549. Compared with PSTG1, recombinant PSTG2 more specifically acted on the β-1,2-glucosidic linkage of the STG molecule to transiently accumulate a larger amount of 6-O-(β-D-glucopyranosyl)-β-D-glucopyranosylsesaminol.     

人参皂甙Rb1,Rg1,Re和Rh1对细胞脱氢酶活性的影响

应用显微分光光度术,定量地分析了人参皂甙Rb_1、Rg_1、Re、Rh_1对人胚肺成纤维细胞(2BS)和HeLa细胞脱氢酶活性的影响。结果表明,4种单体皂甙增加了高代龄2BS细胞内乳酸脱氢酶(LDH),琥珀酸脱氢酶(SDH),葡萄糖-6-磷酸脱氢酶(G-6-PDH)和丙酮酸脱氢酶(PVO)的活性,降低了HeLa细胞内这几种酶的活性。     

Conversion of major ginsenoside Rb1 to 20(S)-ginsenoside Rg3 by Microbacterium sp. GS514

Ginseng saponin, the most important secondary metabolite in ginseng, has various pharmacological activities. Many studies have been directed towards converting major ginsenosides to the more active minor ginsenoside, Rg3. Due to the difficulty in preparing ginsenoside Rg3 enzymatically, the compound has been mainly produced by either acid treatment or heating. A microbial strain GS514 was isolated from soil around ginseng roots in a field and used for enzymatic preparation of the ginsenoside Rg3. Blast results of the 16S rRNA gene sequence of the strain GS514 established that the strain GS514 belonged to the genus Microbacterium. Its 16S rRNA gene sequence showed 98.7%, 98.4% and 96.1% identity with those of M. esteraromaticum, M. arabinogalactanolyticum and M. lacticum. Strain GS514 showed a strong ability to convert ginsenoside Rb1 or Rd into Rg3. Enzymatic production of Rg3 occurred by consecutive hydrolyses of the terminal and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1 showing the biotransformation pathway: Rb1-->Rd-->Rg3.     

A novel ginsenosidase from an Aspergillus strain hydrolyzing 6-O-multi-glycosides of protopanaxatriol-type ginsenosides, named ginsenosidase type IV

Herein, a novel ginsenosidase, named ginsenosidase type IV, hydrolyzing 6-O-multi-glycosides of protopanaxatrioltype ginsenosides (PPT), such as Re, R1, Rf, and Rg2, was isolated from the Aspergillus sp. 39g strain, purified, and characterized. Ginsenosidase type IV was able to hydrolyze the 6-O-alpha-L-(1-->2)-rhamnoside of Re and the 6-O-beta-D- (1-->2)-xyloside of R1 into ginsenoside Rg1. Subsequently, it could hydrolyze the 6-O-beta-D-glucoside of Rg1 into F1. Similarly, it was able to hydrolyze the 6-O-alpha-L-(1-->2)- rhamnoside of Rg2 and the 6-O-beta-D-(1-->2)-glucoside of Rf into Rh1, and then further hydrolyze Rh1 into its aglycone. However, ginsenosidase type IV could not hydrolyze the 3-O- or 20-O-glycosides of protopanaxadioltype ginsenosides (PPD), such as Rb1, Rb2, Rb3, Rc, and Rd. These exhibited properties are significantly different from those of glycosidases described in Enzyme Nomenclature by the NC-IUBMB. The optimal temperature and pH for ginsenosidase type IV were 40°C and 6.0, respectively. The activity of ginsenosidase type IV was slightly improved by the Mg(2+) ion, and inhibited by Cu(2+) and Fe(2+) ions. The molecular mass of the enzyme, based on SDS-PAGE, was noted as being approximately 56 kDa.     

Cytostatic effects of structurally different ginsenosides on yeast cells with altered sterol biosynthesis and transport

Triterpene glycosides are a diverse group of plant secondary metabolites, consisting of a sterol-like aglycon and one or several sugar groups. A number of triterpene glycosides show membranolytic activity, and, therefore, are considered to be promising antimicrobial drugs. However, the interrelation between their structure, biological activities, and target membrane lipid composition remains elusive. Here we studied the antifungal effects of four Panax triterpene glycosides (ginsenosides) with sugar moieties at the C-3 (ginsenosides Rg3, Rh2), C-20 (compound K), and both (ginsenoside F2) positions in Saccharomyces cerevisiae mutants with altered sterol plasma membrane composition. We observed reduced cytostatic activity of the Rg3 and compound K in the UPC2-1 strain with high membrane sterol content. Moreover, LAM gene deletion reduced yeast resistance to Rg3 and digitonin, another saponin with glycosylated aglycon in the C-3 position. LAM genes encode plasma membrane-anchored StARkin superfamily-member sterol transporters. We also showed that the deletion of the ERG6 gene that inhibits ergosterol biosynthesis at the stage of zymosterol increased the cytostatic effects of Rg3 and Rh2, but not the other two tested ginsenosides. At the same time, in silico simulation revealed that the substitution of ergosterol with zymosterol in the membrane changes the spatial orientation of Rg3 and Rh2 in the membranes. These results imply that the plasma membrane sterol composition defines its interaction with triterpene glycoside depending on their glycoside group position. Our results also suggest that the biological role of membrane-anchored StARkin family protein is to protect eukaryotic cells from triterpenes glycosylated at the C-3 position.     

Cloning,expression and biochemical characterization of a GH1 β-glucosidase from Cellulosimicrobium cellulans

β-Glucosidase plays an important role in the degradation of cellulose. In this study, a novel β-glucosidase ccbgl1b gene for a glycosyl hydrolase (GH) family 1 enzyme was cloned from the genome of Cellulosimicrobium cellulans and expressed in Escherichia coli BL21 cells. The sequence contained an open reading frame of 1494?bp, encoded a polypeptide of 497?amino acid residues. The recombinant protein CcBgl1B was purified by Ni sepharose fastflow affinity chromatography and had a molecular weight of 57?kDa, as judged by SDS-PAGE. The optimum β-glucosidase activity was observed at 55?°C and pH 6.0. Recombinant CcBgl1B was found to be most active against aryl-glycosides p-nitrophenyl-β-D-glucopyranoside (pNPβGlc), followed by p-nitrophenyl-β-D-galactopyranoside (pNPβGal). Using disaccharides as substrates, the enzyme efficiently cleaved β-linked glucosyl-disaccharides, including sophorose (β-1,2-), laminaribiose (β-1,3-) and cellobiose (β-1,4-). In addition, a range of cello-oligosaccharides including cellotriose, cellotetraose and cellopentaose were hydrolysed by CcBgl1B to produce glucose. The interaction mode between the enzyme and the substrates driving the reaction was modelled using a molecular docking approach. Understanding how the GH1 enzyme CcBgl1B from C. cellulans works, particularly its activity against cello-oligosaccharides, would be potentially useful for biotechnological applications of cellulose degradation.     

人参皂苷Rb1、Rg1、Re对白血病细胞株KG1α增殖的影响

目的:探讨人参皂苷Rb1、Rg1、Re对急性髓系白血病细胞株(KG1α)增殖的影响.方法:取对数生长期KG1α细胞,分设人参皂苷Rb1、Rg1、Re组和常规培养组,以MTT比色法检测作用24h、48h、72h时对KG1α细胞增殖抑制作用,并计算Rb1的IC_(50)值,以此浓度为工作浓度,设常规培养组和处理组,台盼蓝计数法观察对KG1α细胞增殖的影响;流式细胞术测定细胞周期分布的变化.结果:MTT、台盼蓝计数显示人参皂苷单体Rb1、Rg1可抑制KG1α细胞增殖,呈浓度依赖性,以Rb1抑制效应最佳,于作用48h抑制率最高.台盼蓝计数显示人参皂苷单体Rb1-120μmol/L作用48h时抑制率达50.22%;流式细胞术结果提示,与对照组比较,Rb1-120μmol/L组G_2/M期KG1α细胞比例增加(P<0.05).结论:Rb1可抑制KG1α细胞体外增殖,其增殖抑制作用与将KG1α细胞阻滞于G_2/M期有关.     

Effects of ginsenosides on glycine receptor alpha1 channels expressed in Xenopus oocytes

Ginsenosides, major active ingredients of Panax ginseng, are known to regulate the excitatory ligand-gated ion channel activity. Recent reports showed that ginsenosides attenuate nicotinic acetylcholine and NMDA receptor channel activity. However, it is not known whether ginsenosides also affect the inhibitory ligand-gated ion channel activity. We investigated the effect of ginsenosides on human glycine alpha1 receptor channel activity expressed in Xenopus oocytes using a two-electrode voltage clamp technique. Treatment of ginsenoside Rf enhances glycine-induced inward peak current (IGly) with dose dependent and reversible manner but ginsenoside Rf itself did not elicit membrane currents. The half-stimulatory concentrations (EC50) of ginsenoside Rf was 49.8 +/- 8.9 microM. Glycine receptor antagonist strychnine completely blocked the inward current elicited by glycine plus ginsenoside Rf. Cl- channel blocker 4,4'-disothiocyanostilbene-2,2'-disulfonic acid (DIDS) also blocked the inward current elicited by glycine plus ginsenoside Rf. We also tested the effect of eight individual ginsenosides (i.e., Rb1, Rb2, Rc, Rd, Re, Rg1, Rg2, and Ro) in addition to ginsenoside Rf. We found that five of them significantly enhanced the inward current induced by glycine with the following order of potency: Rb1 > Rb2 > Rg2 > or = Rc > Rf > Rg1 > Re. These results indicate that ginsenosides might regulate gylcine receptor expressed in Xenopus oocytes and this regulation might be one of the pharmacological actions of Panax ginseng.     

Ginsenoside content and variation among and within American ginseng (Panax quinquefolius L.) populations

The contents of five ginsenosides (Rg1, Re, Rb1, Rc and Rd) were measured in American ginseng roots collected from 10 populations grown in Maryland. Ginsenoside contents and compositions varied significantly among populations and protopanaxatriol (Rg1 and Re) ginsenosides were inversely correlated within root samples and among populations. The most abundant ginsenoside within a root and by population was either Rg1 or Re, followed by Rb1. Ginseng populations surveyed grouped into two chemotypes based on the relative compositions of Rg1 and Re. Four populations, including the control population in which plants were grown from TN and WI seed sources, contained roots with the recognized chemotype for American ginseng of low Rg1 composition relative to Re. The remaining 6 populations possessed roots with a distinctive chemotype of high relative Rg1 to Re compositions. Chemotype did not vary by production type (wild versus cultivated) and roots within a population rarely exhibited chemotypes different from the overall population chemotype. These results provide support for recent evidence that relative Rg1 to Re ginsenoside contents in American ginseng roots vary by region and that these differences are likely influenced more by genotype than environmental factors. Because the physiological and medicinal effects of different ginsenosides differ and can even be oppositional, our findings indicate the need for fingerprinting ginseng samples for regulation and recommended usage. Also, the High Rg1/Low Re chemotype discovered in MD could potentially be used therapeutically for coronary health based on recent evidence of the positive effects of Rg1 on vascular growth.