• Journal of Ginseng Research
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• v.38 no.4
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• pp.264-269
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• 2014

Background: In this study, we examined the effects of various enzymes on chemical conversions of ginsenosides in ginseng extract prepared by amylases. Methods: Rapidase, Econase CE, Viscozyme, Ultraflo L, and Cytolase PCL5 were used for secondary enzymatic hydrolysis after amylase treatment of ginseng extract, and ginsenoside contents, skin permeability, and chemical compositions including total sugar, acidic polysaccharide, and polyphenols were determined on the hydrolyzed ginseng extract. Results: Rapidase treatment significantly elevated total ginsenoside contents compared with the control (p < 0.05). In particular, deglycosylated ginsenosides including Rg3, which are known as bioactive compounds, were significantly increased after Rapidase treatment (p < 0.05). The Rapidase-treated group also increased the skin permeability of polyphenols compared with the control, showing the highest level of total sugar content among the enzyme treatment groups. Conclusion: This result showed that Rapidase induced the conversion of ginsenoside glycosides to aglycones. Meanwhile, Cytolase PCL5 and Econase treatments led to a significant increase of uronic acid (acidic polysaccharide) level. Taken together, our data showed that the treatments of enzymes including Rapidase are useful for the conversion and increase of ginsenosides in ginseng extracts or products.

• Journal of Pharmacopuncture
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• v.23 no.2
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• pp.79-87
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• 2020

Objectives: Ginsenosides found in ginseng, and the hydrolysates derived from their conversion, exhibit diverse pharmacological characteristics [1]. These have been shown to include anti-cancer, anti-angiogenic, and anti-metastatic effects, as well as being able to provide hepatic and neuroprotective effects, immunomodulation, vasodilation, promotion of insulin secretion, and antioxidant activity. Therefore, the purpose of this study was to examine how quickly the ginsenosides decompose and what kinds of degradation products are created under physicochemical processing conditions that don't involve toxic chemicals or other treatments that may be harmful. Methods: The formation of ginsenoside-Rg2 and ginsenoside-Rg3 was examined. These demonstrated diverse pharmacological effects. Results: We also investigated physicochemical factors affecting their conversion. The heating temperatures and times yielding the highest concentration of ginsenosides (-Rb1, -Rb2, -Rc, -Rd, -Rf, -Rg1, and -Re) were examined. Additionally, the heating temperatures and rates of conversion of these ginsenosides into new 'ginseng saponins', were examined. Conclusion: In conclusion, obtained provide us with effective technology to control the concentration of both ginsenosides and the downstream converted saponins (ginsenoside-Rg2, Rg3, Rg5, and Rk1 etc.), as well as identifying the processing conditions which enable an enrichment in concentration of these compounds.

• Journal of Ginseng Research
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• v.22 no.3
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• pp.205-210
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• 1998

Glucosidic bonds at the C20 position of the sapogenins were hydrolyzed easily in the lower pH, higher temperatures and longer times to give prosapogenins and sugars. The glucosidic bond of saponin at the C3 of ginsenoside-Rb1, which is secondary carbon, was relatively stable due to the low electron density of -0.2. But the bond of saponin at the C20 position, which is tertiary carbon with the relatively high electron density of -0.3, was liable to be hydrolyzed even in weakly acidic solution by the increase of heating time. On the other hand, red ginseng contained 13.34 mg/g of citric acid, 8.78 mg/g of malonic acid, 3.70 mg/g of oxalic acid, 2.13 mg/g of malic acid and 0.44 mg/g of succinct acid. Ginseng saponins were very stable in ginseng extract neutralized with sodium carbonate or sodium bicarbonate corresponding to the equivalent amount of the total organic acid in the red ginseng.

• Journal of Ginseng Research
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• v.39 no.3
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• pp.221-229
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• 2015

Background: Minor ginsenosides, those having low content in ginseng, have higher pharmacological activities. To obtain minor ginsenosides, the biotransformation of American ginseng protopanaxadiol (PPD)-ginsenoside was studied using special ginsenosidase type-I from Aspergillus niger g.848. Methods: DEAE (diethylaminoethyl)-cellulose and polyacrylamide gel electrophoresis were used in enzyme purification, thin-layer chromatography and high performance liquid chromatography (HPLC) were used in enzyme hydrolysis and kinetics; crude enzyme was used in minor ginsenoside preparation from PPD-ginsenoside; the products were separated with silica-gel-column, and recognized by HPLC and NMR (Nuclear Magnetic Resonance). Results: The enzyme molecular weight was 75 kDa; the enzyme firstly hydrolyzed the C-20 position 20-O-${\beta}$-D-Glc of ginsenoside Rb1, then the C-3 position 3-O-${\beta}$-D-Glc with the pathway $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}C-K$. However, the enzyme firstly hydrolyzed C-3 position 3-O-${\beta}$-D-Glc of ginsenoside Rb2 and Rc, finally hydrolyzed 20-O-L-Ara with the pathway $Rb2{\rightarrow}C-O{\rightarrow}C-Y{\rightarrow}C-K$, and $Rc{\rightarrow}C-Mc1{\rightarrow}C-Mc{\rightarrow}C-K$. According to enzyme kinetics, $K_m$ and $V_{max}$ of Michaelis-Menten equation, the enzyme reaction velocities on ginsenosides were Rb1 > Rb2 > Rc > Rd. However, the pure enzyme yield was only 3.1%, so crude enzyme was used for minor ginsenoside preparation. When the crude enzyme was reacted in 3% American ginseng PPD-ginsenoside (containing Rb1, Rb2, Rc, and Rd) at $45^{\circ}C$ and pH 5.0 for 18 h, the main products were minor ginsenosides C-Mc, C-Y, F2, and C-K; average molar yields were 43.7% for C-Mc from Rc, 42.4% for C-Y from Rb2, and 69.5% for F2 and C-K from Rb1 and Rd. Conclusion: Four monomer minor ginsenosides were successfully produced (at low-cost) from the PPD-ginsenosides using crude enzyme.

• Journal of Ginseng Research
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• v.43 no.1
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• pp.95-104
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• 2019

Background: Oxidized low-density lipoprotein (ox-LDL) causes vascular endothelial cell inflammatory response and apoptosis and plays an important role in the development and progression of atherosclerosis. Ginsenoside compound K (CK), a metabolite produced by the hydrolysis of ginsenoside Rb1, possesses strong anti-inflammatory effects. However, whether or not CK protects ox-LDL-damaged endothelial cells and the potential mechanisms have not been elucidated. Methods: In our study, cell viability was tested using a 3-(4, 5-dimethylthiazol-2yl-)-2,5-diphenyl tetrazolium bromide (MTT) assay. Expression levels of interleukin-6, monocyte chemoattractant protein-1, tumor necrosis factor-${\alpha}$, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1 were determined by enzyme-linked immunosorbent assay and Western blotting. Mitochondrial membrane potential (${\Delta}{\Psi}m$) was detected using JC-1. The cell apoptotic percentage was measured by the Annexin V/ propidium iodide (PI) assay, lactate dehydrogenase, and caspase-3 expression. Apoptosis-related proteins, nuclear factor $(NF)-{\kappa}B$, and mitogen-activated protein kinases (MAPK) signaling pathways protein expression were quantified by Western blotting. Results: Our results demonstrated that CK could ameliorate ox-LDL-induced human umbilical vein endothelial cells (HUVECs) inflammation and apoptosis, $NF-{\kappa}B$ nuclear translocation, and the phosphorylation of p38 and c-Jun N-terminal kinase (JNK). Moreover, anisomycin, an activator of p38 and JNK, significantly abolished the anti-apoptotic effects of CK. Conclusion: These results demonstrate that CK prevents ox-LDL-induced HUVECs inflammation and apoptosis through inhibiting the $NF-{\kappa}B$, p38, and JNK MAPK signaling pathways. Thus, CK is a candidate drug for atherosclerosis treatment.

• Journal of Ginseng Research
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• v.39 no.4
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• pp.392-397
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• 2015

Background: Red ginseng (RG) is processed from Panax ginseng via several methods including heat treatment, mild acid hydrolysis, and microbial conversion to transform the major ginsenosides into minor ginsenosides, which have greater pharmaceutical activities. During the fermentation process using microbial strains in a machine for making red ginseng, a change of composition occurs after heating. Therefore, we confirmed that fermentation had occurred using only microbial strains and evaluated the changes in the ginsenosides and their chemical composition. Methods: To confirm the fermentation by microbial strains, the fermented red ginseng was made with microbial strains (w-FRG) or without microbial strains (n-FRG), and the fermentation process was performed to tertiary fermentation. The changes in the ginsenoside composition of the self-manufactured FRG using the machine were evaluated using HPLC, and the 20 ginsenosides were analyzed. Additionally, we investigated changes of the reducing sugar and polyphenol contents during fermentation process. Results: In the fermentation process, ginsenosides Re, Rg1, and Rb1 decreased but ginsenosides Rh1, F2, Rg3, and Compound Y (C.Y) increased in primary FRG more than in the raw ginseng and RG. The content of phenolic compounds was high in FRG and the highest in the tertiary w-FRG. Moreover, the reducing sugar content was approximately three times higher in the tertiary w-FRG than in the other n-FRG. Conclusion: As the results indicate, we confirmed the changes in the ginsenoside content and the role of microbial strains in the fermentation process.

• Proceedings of the Ginseng society Conference
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• 2002.10a
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• pp.66-83
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• 2002

Recently, we have provided evidence that ginsenosides, the active components of Panax ginseng, utilize pertussis toxin (PTX)-insensitive $G{\alpha}_{q/11}-phospholipase\;C-{\beta}3(PLC-{\beta}3)$ signal transduction pathway for the enhancement of $Ca^{2+}-activated\;Cl^{-}$ current in the Xenopus oocyte (British J. Pharmacol. 132, 641-647, 2001; JBC 276, 48797-48802, 2001). Other investigators have shown that stimulation of receptors linked to $G{\alpha}-PLC$ pathway inhibits the activity of G proteincoupled inwardly rectifying $K^+$ (GIRK) channel. In the present study, we sought to determine whether ginsenosides influenced the activity of GIRK 1 and GIRK 4 (GIRK 1/4) channels expressed in the Xenopus oocyte, and if so, the underlying signal transduction mechanism. In oocyte injected with GIRK 1/4 channel cRNAs, bath-applied ginsenosides inhibited high potassium (HK) solution-elicited GIRK current $(EC_{50}:4.9{\pm}4.3\;{\mu}g/ml).$ Pretreatment of the oocyte with PTX reduced the HK solution-elicited GIRK current by $49\%,$ but it did not alter the inhibitory ginsenoside effect on GIRK current. Prior intraoocyte injection of cRNA(s) coding $G{\alpha}_q,\;G{\alpha}_{11}\;or\;G{\alpha}_q/G{\alpha}_{11},\;but\;not\;G{\alpha}_{i2}\;or\;G{\alpha}_{oA}$ attenuated the inhibitory ginsenoside effect. Injection of cRNAs coding $G{\beta}_{1{\gamma}2}$ also attenuated the ginsenoside effect. Similarly, injection of the cRNAs coding regulators of G protein signaling 1, 2 and 4 (RGS1, RGS2 and RGS4), which interact with $G{\alpha}_i\;and/or\;G{\alpha}_{q/11}$ and stimulates the hydrolysis of GTP to GDP in active GTP-bound $G{\alpha}$ subunit, resulted in a significant reduction of ginsenoside effect on GIRK current. Preincubation of GIRK channel-expressing oocyte in PLC inhibitor (U73122) or protein kinase C (PKC) inhibitor (staurosporine or chelerythrine) blocked the inhibitory ginsenoside effect on GIRK current. On the other hand, intraoocyte injection of BAPTA, a free $Ca^{2+}$ chelator, had no significant effect on the ginsenoside action. Taken together, these results suggest that ginsenosides inhibit the activity of GIRK 1/4 channel expressed in the Xenopus oocyte through a PTX-insensitive and $G{\alpha}_{q/11}$-,PLC-and PKC-mediated signal transduction pathway.

• Journal of Food Hygiene and Safety
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• v.36 no.5
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• pp.418-429
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• 2021

This paper investigates the antioxidant activity and quality characteristics of yanggaeng containing white ginseng and red ginseng extracts and their enzyme hydrolysates that were produced for the purpose of the study. White and red ginseng extracts were hydrolyzed using Rapidase C80 max, Pyr-flo, and Ultimase MFC. Ginsenoside F2 and compound K (CK) were not detected in white and red ginseng before enzymic reaction but were detected in white and red ginseng hydrolyzed through Rapidase C80 max, Pyr-flo, and Ultimase MFC, and the content of CK was the highest in the second enzymic reaction group of red ginseng. Upon preparing yanggaeng containing white and red ginseng before or after enzymatic hydrolysis, the polyphenol content and antioxidant abilities were analyzed. The yanggaeng containing enzyme-hydrolyzed white ginseng and red ginseng showed greater total polyphenol content, superior DPPH radical scavenging activity, superior ABTS radical scavenging activity, and superior FRAP analysis results compared to the yanggaeng that doesn't contain white or red ginseng. As the enzymic reaction was performed in the added white and red ginseng, the antioxidant activity increased significantly (P<0.05). In brightness(L^*), non-additive yanggaeng (control group) was the highest, red ginseng yanggaeng (RG) showed the highest redness(a^*), and the white ginseng yanggaeng (WG) showed the highest yellowness(b^*). In terms of texture, the yanggaeng containing red ginseng with second hydrolysis (RG-T2) showed significantly high results in hardness, springiness, chewiness, cohesiveness, and gumminess (P<0.05). In conclusion, treating white and red ginseng with Rapidase C80 max, Pyr-flo, and Ultimase MFC is very useful in ginsenoside deglycosylation and will produce CK with excellent biological activity. It can also be seen that yanggaeng containing white and red ginseng hydrolyzed with enzymes significantly increase total polyphenol and antioxidant activity compared to the control group (yanggaeng with no added ginseng). These results will be useful as excellent foundational data for the production of functional yanggaeng in the future.

• Journal of Ginseng Research
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• v.24 no.3
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• pp.153-156
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• 2000

In order to improve the qualities of red ginseng extract and decrease precipitate formation in ginseng drink, red ginseng extract were hydrolyzed with ${\alpha}$-Amylase and characteristics of the hydrolyzed ginseng extract were investigated. 1.08% of isomaltose were produced and glucose content was increased from 2.83% to 11.03% in the hydrolyzed red ginseng extract. Total ginsenoside content of the hydrolyzed ginseng extract were decreased from 1,661 mg/100g extract to 1,389 mg/100g extract. The hydrolyzed ginseng extract enhanced the growth of Lactobacillus casei, Lactobacillus rhamnosus and Lactobacillus helveticus. Bitterness and astringency of the hydrolyzed ginseng extract were lower than those of the ginseng extract Precipitate formations in ginseng drink prepared with the hydrolyzed ginseng extract were significantly reduced in the storage conditions of 40$^{\circ}C$ for 4 weeks compared to those of control.

• Journal of Ginseng Research
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• v.6 no.1
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• pp.25-29
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• 1982

This investigation was carried out to study the influence of pH and heat treatment on the ginsenosides in the white ginseng extract. Changes in ginsenosides (Rb1, Rb2, ,Rc, Rd) and free sugar were measured by the peak area variation of HPLC chromatogram during 25 hours heat treatment at the various level of pH. It was found that :(1) The peak areas of Rb1. Rb2, Rc and Rd on the HPLC chromatogram were decreased remarkably below pH 4.0 and more decrease was found as the temperature and heating time increased. (2) Those of glucose and arabinose were increased remarkably. It is considrered that the increase of glucose and the formation of arabinose result from the hydrolysis of ginsenoside( Rb1, Rb2, Rc, Rd) linked with sugars.