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Korean Red Ginseng
KOREAN RED GINSENG is steamed and dried with the skin unpeeled
from the strictly selected fresh ginseng, Panax ginseng C.A. Meyer. It
is in light yellowish brown or light reddish brown color. It may be
preserved for a long time of 10 years or more. Korean ginseng is well
known for its rich saponin content called Ginsenoside that activates
the functions of human body's components as well as strengthens
immunity. The six-year Korean red ginseng root contains the most
saponin among worldwide ginsengs and the composition is so
exquisite as to display high efficacy and properties.
WHITE GINSENG
RED GINSENG
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Fermented Korean Red Ginseng
It is known that the active components of Ginseng, ginsenosides are
hydrolyzed by intestinal bacteria and then the metabolites are absorbed into
the body. But the ginsenoside-hydrolyzing ability of intestinal bacteria varies
from person to person by person's physical health and the amount and type of
intestinal bacteria that exists in the intestines.
During fermentation, special enzymes are formed to break down not only the
ginsenosides but also other important ingredients of Red Ginseng, thus
making it easy for everyone to absorb ingredients of ginseng (It has already
been pre-hydrolyzed by fermentation)
BIO-FRGE is a fermented Korean Red Ginseng Extract produced by Bioland ‘s
high fermentation technology to increase absorption, reduce bitter taste, and
also increase anti-aging property of Red Ginseng.
Extraction
&
Fermentation
RED GINSENG
BIO-FRGE
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Metabolism of Ginsenoside Rb1
Ginsenosides Contents of BIO-FRGE
9
Red Ginseng
8
Fermented Red Ginseng
Contents (mg/g)
7
6
5
4
3
2
1
Rk1
0
Rg1
Rb1
Rg3
Rk1
Ginsenosides
Rd
Rg3-S
Rg3-R
Rk1
Rg5
Total
in BIO-FRGE
Content
(mg/g)
0.11
0.27
0.10
0.15
0.17
3.50
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Cytotoxicity of BIO-FRGE
® Cytotoxicity (MTT method)
Cytotoxicity of FRGE by MTT assay.
sample
MTT
treatment
treatment
Incubation
Absorbance
measurement
(570nm)
Incubation
Incubation
DMSO
treatment
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Cytotoxicity of BIO-FRGE
® Result (human skin fibroblast cell
(ATCC, CRL-2076))
110
100
90
cell viability (%)
80
70
60
50
40
30
20
10
0
10
25
50
100
250
conc. (ug/ml)
Fig 1. Cytotoxicity assay of FRGE. The data were expressed as mean values
(± standard deviations) of five experiments.
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Cytotoxicity of BIO-FRGE
® Result (human skin keratinocyte
(HaCaT))
110
100
cell viability (%)
90
80
70
60
50
40
30
20
10
0
1
10
25
concentration (ug/ml)
50
100
Fig 2. cell proliferation assay of FRGE. The data were expressed as mean
values (± standard deviations) of five experiments.
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Anti-aging of BIO-FRGE
® SIRTUIN
-It’s looking like these sirtuins serve as
guardians of the cell. These enzymes allow
cells to survive damage and delay cell death.
– David Sinclair / Harvard Medical School
researcher
Crystallographic structure of yeast sir2 (rainbow colored
-Sirtuins are the central regulator of the aging
process. – Leonard Guarente / Massachusetts
Institute of Technology (MIT) in Cambridge
cartoon, N-terminus = blue, C-terminus = red) complexed with
ADP (space-filling model, carbon = white, oxygen = red,
nitrogen = blue, phosphorous = orange) and a histone H4
peptide (magenta) containing an acylated lysine residue
(displayed as spheres)
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Anti-aging of BIO-FRGE
®
®
Silent Information Regulator 2 (Sir2) proteins (sirtuins)
ƒ
A class of proteins which possess either histone deacetylase or mono-ribosyltransferase activity
and are found in organisms ranging from bacteria to humans.
ƒ
Yeast Sir2 and some, but not all, sirtuins are protein deacetylases. Unlike other known protein
deacetylases, which simply hydrolyze acetyl-lysine residues, the sirtuin-mediated deacetylation
reaction couples lysine deacetylation to NAD hydrolysis. This hydrolysis yields O-acetyl-ADPribose, the deacetylated substrate and nicotinamide, itself an inhibitor of sirtuin activity. The
dependence of sirtuins on NAD links their enzymatic activity directly to the energy status of the
cell via the cellular NAD:NADH ratio, the absolute levels of NAD, NADH or nicotinamide or a
combination of these variables.
ƒ
Sirtuins have been implicated in influencing aging and regulating transcription, apoptosis and
stress resistance.
Species distribution
ƒ
Whereas bacteria and archaea encode either one or two sirtuins, eukaryotes encode several
sirtuins in their genomes. In yeast, roundworms, and fruit flies sir2 is the name of the sirtuin-type
protein. This research started in 1991 by Leonard Guarente of MIT. Mammals possess seven
sirtuins (SIRT1-7) that occupy different subcellular compartments such as the nucleus (SIRT1, 2, -6, -7), cytoplasm (SIRT1 and SIRT2) and the mitochondria (SIRT3, -4 and -5).
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Anti-aging of BIO-FRGE
® Types
ƒ Sirtuins are classed according to their sequence of amino acids. Prokaryotics are in class U.
ƒ In yeast (a lower eukaryote), sirtuin was initially found and named sir2. In more complex mammals there
are seven known enzymes which act as on cellular regulation as sir2 does in yeast.
ƒThese genes are designated as belonging to different classes, depending on their amino acid sequence
structure
Species
Yeast
Mouse
Human
Intracellular
location
Sir2 or Sir2p,
Hst1 or Hst1p
Sirt1
SIRT1
nucleus
deacetylase
metabolism
inflammation
Hst2 or Hst2p
Sirt2
SIRT2
cytoplasm
deacetylase
cell cycle
tumorigenesis
Sirt3
SIRT3
nucleus and
mitrochondira
deacetylase
metabolism
II
Sirt4
SIRT4
mitochondria
ADP-ribosyl
transferase
insulin secretion
III
Sirt5
SIRT5
mitochondria
deacetylase
unknown
a
Sirt6
SIRT6
nucleus
ADP-ribosyl
transferase
DNA repair
b
Sirt7
SIRT7
nucleus
unknown
rDNA
transcription
regulation of
acetyl-CoA synthetase
metabolism
Class
Subclass
Bacteria
a
I
Activity
Function
b
c
Hst3 or Hst3p,
Hst4 or Hst4p
IV
U
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Anti-aging of BIO-FRGE
The yeast sirtuin (Sir2)
is a histone deacetylase
that modulates yeast
replicative life span by
suppressing genome
instability through
chromatin modification.
Oberdoerffer et al. (2008)
report that SIRT1, the
mammalian ortholog of
Sir2, is involved in DNA
damage-induced chromatin
reorganization, which
promotes genome stability
in mammalian cells.
Anti-aging of BIO-FRGE
Anti-aging of BIO-FRGE
An increasing dose of Sir2 extends yeast
replicative life span,
whereas its loss reduces natural longevity
overexpression of Sir2 orthologs in
nematodes and flies also extends life span.
In diverse organisms, Sir2 slows the pace
of ageing and increases maximum lifespan.
Scientists show that the potent activator resveratrol, a polyphenol found in red wine,
increases DNA stability and lifespan by 70% by stimulating Sir2. Nature 425, 191 (2003).
Hypothesis: A potent activator of Sir2 like resveratrol would be helpful as an anti-aging
ingredient in cometics.
Anti-aging of BIO-FRGE
® SIRT3
ƒ Sirtuin (silent mating type information regulation 2 homolog) 3 (S. cerevisiae),
is the third member of the mammalian sirtuin family. The SIRT3 gene encodes the
protein SIRT3, which exhibits NAD+-dependent deacetylase activity.
ƒ SIRT3 is a member of the sirtuin family of proteins, homologs to the yeast Sir2
protein. Members of the sirtuin family are characterized by a sirtuin core domain
and grouped into four classes and the protein encoded by this gene is included in
class I of the sirtuin family. The human sirtuins have a range of molecular
functions and have emerged as important proteins in aging, stress resistance
and metabolic regulation. In addition to protein deacetylation, studies suggest
that the human sirtuins may function as intracellular regulatory proteins with mono
ADP ribosyltransferase activity.
ƒ Structure
While the crystal structure has not yet been solved, SIRT3 is a soluble protein
located in the mitochondrial matrix, and contains a mitochondrial processing
peptide at the n-terminus.
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Anti-aging of BIO-FRGE
® Function
- Mitochondrial
ƒ Three sirtuins, SIRT3, SIRT4 and SIRT5, are located in mitochondria and have been implicated
in regulating metabolic processes. Endogenous SIRT3 is a soluble protein located in the
mitochondrial matrix. Overexpression of Sirt3 in cultured cells increases respiration and
decreases the production of reactive oxygen species. Fasting increases Sirt3 expression in
white and brown adipose tissue (WAT and BAT, respectively) and overexpression of SIRT3 in
HIB1B brown adipocytes increases the expression of PGC-1α and UCP1, suggesting a role for
SIRT3 in adaptive thermogenesis BAT. BAT is different from WAT because it harbors large
numbers of mitochondria and is important for thermogenesis in rodents. Thermogenesis in BAT
is mediated by the uncoupling protein 1 (UCP1), which induces proton leakage and thereby
generates heat instead of ATP. Mechanistic insights into how SIRT3 affects thermogenesis in
BAT is lacking and whether SIRT3 affects UCP1 activity directly is not known.
ƒ In addition to controlling metabolism at the transcriptional level, sirtuins also directly control the
activity of metabolic enzymes. In Salmonella enterica, the bacterial sirtuin CobB regulates the
activity of the enzyme acetyl-coenzyme A (acetyl-CoA) synthetase. As mentioned above,
orthologs of acetyl-CoA synthetase exist in the cytoplasm (AceCS1) and in mitochondria
(AceCS2) in mammals. The presence of the sirtuin deacetylase SIRT3 in the mitochondrial
matrix suggests the existence of lysine acetylated mitochondrial proteins. Indeed, SIRT3
deacetylates and activates the mammalian mitochondrial acetyl-coA synthetase (AceCS2).
Furthermore, SIRT3 and AceCS2 are found complexed with one another, suggesting a critical
role for control of AceCS2 activity by SIRT3.
Anti-aging of BIO-FRGE
® Nuclear
ƒ In addition to its reported mitochondrial function, some researchers have proposed a
very small pool of active nuclear SIRT3 exists. This pool is reported to consist of the
long form of SIRT3 and has been suggested to have histone deacetylase activity.
The observation that SIRT3 has nuclear activity came from a report that SIRT3
protected cardiomyocytes from stress mediated cell death and that this effect was
due to deacetylation of a nuclear factor,Ku-70.
® Clinical significance
ƒ The is a strong association between SIRT3 alleles and longevity in males
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Anti-aging of BIO-FRGE
® Sirt 3 expression activity (RT-PCR)
Measuring Sirt 3 expression level in the human skin fibroblast cell
(ATCC, CRL-2076).
sample
treatment
treatment
Incubation
Incubation
Electrophoresis
RT-PCR
RNA isolation
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Anti-aging of BIO-FRGE
Effect of BIO-FRGE on the expression of Sirt 3.
Control
Resveratrol
BIO-FRGE
200
% of control
150
100
50
0
Control (FBS 10%)
Concentration (µg/ml)
Resveratrol 10 uM
FRGE 10 ug/ml
FRGE 50 ug/ml
FRGE 100 ug/ml
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Anti-aging of BIO-FRGE
- Inflammation and aging
- Reference from : Chung HY, Cesari M, Anton S, Marzetti E, Giovannini S, Seo AY, Carter C, Yu BP,
Leeuwenburgh C (2009) Molecular inflammation: underpinnings of aging and age-related diseases. Ageing
Res Rev, 8, 18-30.
Anti-aging of BIO-FRGE
RNS inhibition
ROS inhibition
iNOS expression rate (%)
100
100
80
60
40
28.6
17.79
20
5.76
0.82
C O X - 2 e x p r e s s i o n r a te ( % )
120
120
100
100
80
60
48.79
43.09
48.21
40
20
3
0
0
10
Control
LPS
100
10
250
BIO-FRGE (㎍/㎖)
iNOS mRNA Expression inhibition
Control
LPS
100
250
BIO-FRGE (㎍/㎖)
COX-2 mRNA Expression inhibition
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