This thesis is about some properties of the enzyme conversed - saponin containing Compound K.
MFDS merged most kinds of ginseng products (Red ginseng, China ginseng, Cultivated wild ginseng, Cultivated ginseng, etc) into one as ‘the Ginseng’ in 2014 so it should be the time to develop those pro...
This thesis is about some properties of the enzyme conversed - saponin containing Compound K.
MFDS merged most kinds of ginseng products (Red ginseng, China ginseng, Cultivated wild ginseng, Cultivated ginseng, etc) into one as ‘the Ginseng’ in 2014 so it should be the time to develop those products with high-quality and highly-safe to make them more beneficial.
To investigate whether other parts of ginseng body except roots are capable to develop as valuable source materials. Because, the use of those leaves and stems are rarely known, whilst the roots of ginseng has been traditionally used in common. However, The Ames test, a test of bacterial reverse mutation to see genetical toxicity, showed some toxicities with the 95% ethanol-extracts from leaves and stems so the use of these parts as food materials is not recommendable unless the safety is proven in future.
To develop new products, another trial with ginseng roots has been conducted to activate source materials with heating. Activation of saponin has been known to increase the rate of body uptake when it dosed in human and to elevate its efficacy by dissociating those saccharides from saponin. Likely to the traditional process of red ginseng production, heating at 95 ℃ for 30-40 mins or at 90 ℃ for 4-6 hrs can derive similar outcomes.
To make higher efficacy of the products, the previous activation process with heating twice at 121 ℃, 15 Lb/cm2 for 15 mim was modified so heat distribution of the ginseng become homogeneous as to increase its uniformity by pre-heating at 80 ℃ for 1~2 hrs. After the pre-heat treatment, in order of processes with a high temp-steam sterilizer at 100 ℃ for 30 min, at 121 ℃ for 15min, cooling at room temp, and then re-heat at 121 ℃ for 15min or three times of repeated lower-temp heating process at 110℃ for 10 min, as a final process, was used so the activated form of ginseng sample was produced.
For the enzymatic conversion to CKS, Rapidase TF, one of enzymes, like polygalacturonase and pectinase, which contains more than 45% of glycerol and was used as stabilizer. Actigin was used as a source material of enzymatic conversion in orders of extraction with an 80% of ethanol, condensation, and dilution processes.
Higher recovery rate of CKS was obtained by a series of procedures which are injection of an 8 ml/min of diluted extracts into an enzyme reactor, reaction with the converting enzyme at 50℃ for 10 hrs, and chilling in a refrigerator for 16 hrs. As an achievement of those completely solubilized formulation experiments, the totally soluble CKS was produced. Those major constituents of the CKS solution were compound K, Rd, Rg3, and F2. As those basic amino acids have a tendency to unfold those poorly-soluble substances, the body injectable CKS solution was prepared with a mixture of 1% lysine, 1% tween 80, and 3% ethanol then the pH was adjusted to 8.6. The concentration of stock CKS solution was 10 mg/ml.
To investigate the effect of ginseng activation on the production of acidic polysaccharides. The freeze dried Actigin was powdered and was extracted with 30% ethanol then the supernatant was taken after centrifugation. The precipitate from the 70% supernatant adjusted with ethanol was dissolved into distilled water then dialyzed through a semipermeable membrane with the pore size of 14,000 MWCO. The dialyzed sample was re-adjusted its proportion to 70% then the crude polysaccharide precipitate was obtained after centrifugation, and the production of acidic polysaccharide was found to be increased to 40% more than that with red ginseng. According to the obtained results, the used manufacturing method in this study was supposed to be proper to produce Actigin.
Some of possible pharmacological activities were examined. Increased food intake and physical activity were observed during CKS administration in aged-rats(21 months). The 37 days of intraperitoneal administration of CKS, in comparison to the control animal, showed a 30% increased number of red blood cell and a 44.5% decreased concentration of triglyceride in blood.
The ambivalent characteristics in cells was also observed that as CKS stimulated differentiation of human-origin stem cells while it suppressed cancer cell growth. In the single toxicity test in rats, more than 120 times of CKS dose(15 mg/kg) for 14 days did not show any inordinary signs of toxicity.
Another approach that effects of CKS on life span in blood cells was also studied and the concentration of intracellular β-galactosidase was shown to be decreased to an approximate 20.3% of the control in average value.
The stimulative effect of CKS on cell differentiation was observed as the doubling time of teeth-origin mesenchymal stem cell was 34% shortened. In contrast, the observed 20% longer doubling time of human lung cancer cell (i.e. A549) suggested the suppressive effects on cancer cell growth.
In addition, either CKS dose of 12.5 mg/kg or 25 mg/kg, to see the effects on male productivity, increased both the sperm motility and cell number in rat.
This thesis is about some properties of the enzyme conversed - saponin containing Compound K.
MFDS merged most kinds of ginseng products (Red ginseng, China ginseng, Cultivated wild ginseng, Cultivated ginseng, etc) into one as ‘the Ginseng’ in 2014 so it should be the time to develop those products with high-quality and highly-safe to make them more beneficial.
To investigate whether other parts of ginseng body except roots are capable to develop as valuable source materials. Because, the use of those leaves and stems are rarely known, whilst the roots of ginseng has been traditionally used in common. However, The Ames test, a test of bacterial reverse mutation to see genetical toxicity, showed some toxicities with the 95% ethanol-extracts from leaves and stems so the use of these parts as food materials is not recommendable unless the safety is proven in future.
To develop new products, another trial with ginseng roots has been conducted to activate source materials with heating. Activation of saponin has been known to increase the rate of body uptake when it dosed in human and to elevate its efficacy by dissociating those saccharides from saponin. Likely to the traditional process of red ginseng production, heating at 95 ℃ for 30-40 mins or at 90 ℃ for 4-6 hrs can derive similar outcomes.
To make higher efficacy of the products, the previous activation process with heating twice at 121 ℃, 15 Lb/cm2 for 15 mim was modified so heat distribution of the ginseng become homogeneous as to increase its uniformity by pre-heating at 80 ℃ for 1~2 hrs. After the pre-heat treatment, in order of processes with a high temp-steam sterilizer at 100 ℃ for 30 min, at 121 ℃ for 15min, cooling at room temp, and then re-heat at 121 ℃ for 15min or three times of repeated lower-temp heating process at 110℃ for 10 min, as a final process, was used so the activated form of ginseng sample was produced.
For the enzymatic conversion to CKS, Rapidase TF, one of enzymes, like polygalacturonase and pectinase, which contains more than 45% of glycerol and was used as stabilizer. Actigin was used as a source material of enzymatic conversion in orders of extraction with an 80% of ethanol, condensation, and dilution processes.
Higher recovery rate of CKS was obtained by a series of procedures which are injection of an 8 ml/min of diluted extracts into an enzyme reactor, reaction with the converting enzyme at 50℃ for 10 hrs, and chilling in a refrigerator for 16 hrs. As an achievement of those completely solubilized formulation experiments, the totally soluble CKS was produced. Those major constituents of the CKS solution were compound K, Rd, Rg3, and F2. As those basic amino acids have a tendency to unfold those poorly-soluble substances, the body injectable CKS solution was prepared with a mixture of 1% lysine, 1% tween 80, and 3% ethanol then the pH was adjusted to 8.6. The concentration of stock CKS solution was 10 mg/ml.
To investigate the effect of ginseng activation on the production of acidic polysaccharides. The freeze dried Actigin was powdered and was extracted with 30% ethanol then the supernatant was taken after centrifugation. The precipitate from the 70% supernatant adjusted with ethanol was dissolved into distilled water then dialyzed through a semipermeable membrane with the pore size of 14,000 MWCO. The dialyzed sample was re-adjusted its proportion to 70% then the crude polysaccharide precipitate was obtained after centrifugation, and the production of acidic polysaccharide was found to be increased to 40% more than that with red ginseng. According to the obtained results, the used manufacturing method in this study was supposed to be proper to produce Actigin.
Some of possible pharmacological activities were examined. Increased food intake and physical activity were observed during CKS administration in aged-rats(21 months). The 37 days of intraperitoneal administration of CKS, in comparison to the control animal, showed a 30% increased number of red blood cell and a 44.5% decreased concentration of triglyceride in blood.
The ambivalent characteristics in cells was also observed that as CKS stimulated differentiation of human-origin stem cells while it suppressed cancer cell growth. In the single toxicity test in rats, more than 120 times of CKS dose(15 mg/kg) for 14 days did not show any inordinary signs of toxicity.
Another approach that effects of CKS on life span in blood cells was also studied and the concentration of intracellular β-galactosidase was shown to be decreased to an approximate 20.3% of the control in average value.
The stimulative effect of CKS on cell differentiation was observed as the doubling time of teeth-origin mesenchymal stem cell was 34% shortened. In contrast, the observed 20% longer doubling time of human lung cancer cell (i.e. A549) suggested the suppressive effects on cancer cell growth.
In addition, either CKS dose of 12.5 mg/kg or 25 mg/kg, to see the effects on male productivity, increased both the sperm motility and cell number in rat.
주제어
#Panax ginseng
#ginsenosides
#Formulation
#Rejuvenation
#RBC
#CKS
#enzyme conversion
#Acidic polysaccharide
#Mesenchymal stem cell
#β-galactosidase
#Doubling time
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