JOURNAL OF MEDICINAL FOOD

J Med Food 17 (1) 2014, 119–127

# Mary Ann Liebert, Inc., and Korean Society of Food Science and Nutrition
DOI: 10.1089/jmf.2013.2877

Doenjang, a Korean Fermented Soy Food, Exerts Antiobesity

and Antioxidative Activities in Overweight Subjects with the PPAR-c2
C1431T Polymorphism: 12-Week, Double-Blind Randomized Clinical Trial
Youn-Soo Cha,1,* Yongsoon Park,2,* Myoungsook Lee,3,* Soo-Wan Chae,4
Kungmin Park,2 Yeonsoo Kim,5 and Haeng-Shin Lee 6
1
Department of Food Science and Human Nutrition, Obesity Research Center; 4Clinical Trial Center for Functional Foods;
Chonbuk National University, Jeonju, Republic of Korea.
2
Department of Food and Nutrition, Hanyang University, Seoul, Republic of Korea.
3
Department of Food and Nutrition, Research Institute of Obesity Sciences, Sungshin Women’s University,
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Seoul, Republic of Korea.

5
Department of Nutrition and Dietetics, School of Human Ecology, Louisiana Tech University, Ruston, Louisiana, USA.
6
Korea Health Industry Development Institute, Osongsaengmyeong-ro, Cheongwon-gun, Chungcheongbukdo, Republic of Korea.

ABSTRACT We examined the antiobesity and antioxidant effects of supplementation with doenjang, a fermented soybean
paste, in overweight Koreans with the PPAR-c2 C1431T polymorphism. Sixty overweight subjects were randomly assigned to
consume either 9.8 g/day of doenjang or placebo for 12 weeks. Before and after the intervention, anthropometric and
metabolic parameters, along with abdominal fat distribution and PPAR-c2 polymorphisms, were measured. Fifty-one subjects
completed the study, doenjang (n = 26) and placebo (n = 25) groups. Relative frequencies of the PPAR-c2 genotypes CC, TC,
and TT were 70% (n = 41), 25.9% (15), and 3.4% (2), whereas those of the PPAR-c2 alleles C and T were 81.6% and 18.4%.
Visceral fat area (VFA) was significantly decreased by doenjang supplementation in subjects with a mutant T allele of PPAR-
c2 compared to those with a C allele after adjusting for age, sex, and body mass index. Plasma free fatty acid, insulin, and
homeostatic model assessment insulin resistance (HOMA-IR) levels were also significantly increased in the doenjang group.
Doenjang pills significantly activated radical clearance capacity (ORAC and DNA tail length) in subjects with the C allele.
The catalase (CAT) activity was increased twofold in the doenjang-treated group with the C allele, but this phenomenon was
reversed in those with the T allele. Doenjang-treated subjects tended to have low dietary carbohydrate and sodium intakes
compared with those given placebo. We found that doenjang supplementation decreased visceral fat accumulation and aging
most effectively in subjects with PPAR-c polymorphisms. This study suggests that doenjang has antiobesity and antioxidative
effects in overweight individuals with mutant alleles of PPAR-c2.

KEY WORDS:  doenjang (fermented soypaste)  soybean  obesity  antioxidant  PPAR-c2 polymorphism

INTRODUCTION foods in the management of obesity has increased, but the

O besity is a complex metabolic disorder characterized
by excessive accumulation of fat in various tissues and
organs as a result of higher energy intake and lower energy
expenditure. The increased incidence of obesity has resulted
in concerted efforts by governments and industries to de-
velop effective interventions to stem the worldwide rise in
obesity rates.1,2 As a result, the importance of functional
*These authors contributed equally to this work.
Manuscript received 18 March 2013. Revision accepted 9 December 2013.
Address correspondence to: Myoungsook Lee, PhD, Department of Food and Nutrition,
Research Institute of Obesity Sciences, Sungshin Women’s University, 249-1, Dongsundong-
3ka, Sungbuk-ku, Seoul 142-732, Republic of Korea, E-mail: drmlee79@gmail.com;
mlee@sungshin.ac.kr
most effective target foods have not yet been systematically
explored.
Soybean, a species of legume native to East Asia, con-
tains significant amounts of phytic acid, a-linolenic acid,
isoflavone, daizein, polyphenols, and saponins, all of which
have many biological functions, such as antioxidant, anti-
cancer, immune strengthening, and antiobesity activity.3,4
The U.S. Food and Drug Administration has approved a
health claim stating that 25 g of soy protein per day protects
against cardiovascular disease.5 Park et al. reported that
Koreans have consumed doenjang soup at least once a day
for hundreds of years.6 Soybean is popular for its functional
ingredients and meat-like texture and flavor, despite a much
lower energy density than meat. Traditional nonfermented
food products made from soybean include soymilk, tofu,
and yuba, whereas the common soy-based fermented foods

119
 120 CHA ET AL.

include soy sauce, doenjang, tempeh, miso, and natto.
Doenjang is a traditional Korean fermented soy bean paste
similar to Japanese miso, which is fermented from meju
using Bacillus and Aspergillus species of bacteria.7,8 Fer-
mentation of soybeans alters the profiles of isoflavonoids
(genistein and daizein), small peptides (Glu-Tyr and Glu-
Phe), and urea cycle intermediates (i.e., arginine as a NO
precursor, citrulline, and ornithine).9,10 Such alterations
have important effects on the prevention of metabolic dis-
eases, including obesity.
Although PPAR-c is well known for its role in adipo-
genesis, it also plays a crucial role in maintaining normal
physiology, including insulin sensitization.11 Mutations in
the PPAR-c gene are associated with obesity and diabetes-
related phenotypes.12 The polymorphism C1431T, located
in exon 6 of PPAR-c, is associated with susceptibility to
cardiovascular diseases,13,14 leptin concentrations,15 and
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supplements during the study. A physical and pathological
questionnaire was administered and anthropometric mea-
surements were made. In addition, all biochemical samples
were collected in accordance with the Ethics Committees of
the CTCF2. However, nine participants, four in the doen-
jang group and five in the placebo group, failed to complete
the study. Seven participants were disqualified because of an
inadequate intake of the prescribed supplements or not
participating in other aspects of the study, while two par-
ticipants voluntarily withdrew. Finally, 51 subjects (doen-
jang = 26, placebo = 25) completed the study after 12 weeks
and statistical analysis was carried out for the final 51 sub-
jects. This study was conducted according to the guidelines
laid down in the Declaration of Helsinki, and all procedures
involving human subjects were approved by the Chonbuk
National University Ethical Board of Clinical Experiments.
Informed and written consent was obtained from all subjects

body mass index (BMI).16–17 before starting the study.

PPAR-c deficiency is associated with a greater risk of ad-
ipose tissue inflammation as well as increased susceptibility Test and placebo food production
to diet-induced obesity.18,19 Previous studies on isoflavones
Doenjang (38.5–40 g; Soon-Chang, Korea) was freeze
have shown their potential antiobesity effects, but the
dried in 9.8 g/3 packs/day (Lymsil Medicine Co., Jeonju,
mechanisms related to PPAR-c are thus far unclear.20,21 A
Korea), which were consumed three separate times per day.
few studies have shown that doenjang cooked in any manner
Doenjang and placebo provided 63 and 61 kcal/day, re-
reduces body fat mass. However, a randomized clinical study
spectively (Table 1). The actual nutrients per 100 kcal of
using doenjang on subjects with C1431T polymorphism of
doenjang were calculated based on the total recommended
the PPAR-c2 gene has not yet been conducted.
intake of about 2100–2600 kcal for adults. The doses were
The goal of this study was to investigate whether sup-
determined from previous animal studies using animal-to-
plementation of fermented soybeans affects body fat dis-
human extrapolation and by using a dose equivalent to
tribution in overweight subjects based on the mechanisms of
approximately three servings per day of doenjang soup,
PPAR-c in a 12-week randomized clinical trial (RCT).
according to Korean FDA policy.
MATERIALS AND METHODS
Anthropometric measurements
Subjects and research design
The waist circumference (WC), blood pressure (BP),
The present 12-week, double-blind, placebo-controlled BMI, and WHR were measured and body fat distribution
RCT followed the Korean FDA’s General Clinical Practice measured using Inbody 3.0 (Biospace Co., Ltd., Seoul,
Guidelines to evaluate the antiobesity and antioxidative Korea) and computed tomography (CT). The visceral fat
effects of doenjang, a Korean traditional fermented soy
paste. The Clinical Trial Center for Functional Food
(CTCF2) at Chonbuk National University Hospital recruited Table 1. Composition of Doenjang and Placebo Supplement
83 participants. Our inclusion criteria were as follows:
adults older than 19 or younger than 65 years, body mass Doenjang Placebo
index (BMI) > 23 kg/m2, and a waist/hip ratio (WHR) Meju 20.0 0.1
> 0.90 for men and > 0.85 for women. Exclusion criteria Salt 3.8 3.5
were the use of any type of medication, in particular, lipid- Water 16.2
lowering and antidiabetic medications, the presence of fa- Cowpea powder 1.3
milial cardiovascular-related diseases or gastrointestinal Wheat flour 0.8
Rice powder 2.8
disorders, or the loss of > 10% of their body weight within Cocoa powder 0.1
the last 3 months. After screening, 60 subjects (10 males and Chrysanthemum tea powder 0.3
50 females, 37.88 – 1.51 years, 69.69 – 1.23 kg) were ran- Oil powder 4.7
domly assigned to either a placebo (n = 30) or doenjang Soy sauce 2.9
group (n = 30). Subjects assigned to the doenjang group took Total weight (g) 40 16.5
9.8 g of freeze-dried doenjang supplement per day for 12 Freeze-dried weight (g) 9.8 9.8
Energy (kcal) 63.9 61.6
weeks, while subjects assigned to the placebo group took a
placebo pill each day for 12 weeks. Subjects were asked to Doenjang was freeze dried in 9.8 g/3 packs/day, which were consumed three
maintain their usual diet and level of physical activity and separate times per day. Doenjang and placebo provided 63 and 61 kcal/day,
were prohibited from consuming functional foods or dietary respectively.
 INTERACTION OF PPAR-c AND FERMENTED SOYBEAN ON OBESITY
area (VFA) was quantified by measuring the intra-abdomi-
nal cavity at the internal aspect of the abdominal and oblique
muscle walls surrounding the cavity and the posterior aspect
of the vertebral body. The subcutaneous fat area (SFA) was
calculated by subtracting the VFA from the total fat area.
Laboratory analysis
Blood was collected after overnight fasting. Whole blood
samples were allocated, allowed to clot, and centrifuged at
1168 g for 15 min to obtain serum. Whole blood and serum
were stored at - 80C for genetic and biochemical analysis.
The following profiles were measured using a Hitachi-7600
analyzer (Hitachi Ltd., Tokyo, Japan): fasting blood sugar,
total cholesterol (TC), triglycerides (TG), high-density li-
poprotein cholesterol (HDL-C), low-density lipoprotein
cholesterol (LDL-C), apoA-I, apo B-100, glucose, insulin,
fructosamin, and hemoglobin A1C (HbA1C). Homeostatic
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model assessment–insulin resistance (HOMA-IR) was cal-
culated using the following formula: fasting insulin (lU/
mL) · fasting glucose (mM)/22.5.22 Hematology parameters
(WBC, RBC, Hb, and platelet) were measured using a
Coulter LH 750 analyzer (Beckman Coulter, Inc., Fullerton,
CA, USA). Aspartate aminotransferase (AST), alanine
aminotransferase (ALT), albumin, blood urea nitrogen
(BUN), creatine, and sodium excretion were measured using
an autoanalyzer (Variant; Bio-Rad, Hercules, CA, USA).
The BP was measured using an Omron T4 (Omron Cor-
poration, Kyoto, Japan).
DNA degradation in lymphocytes was measured by the
comet assay, and the plasma total radical-trapping antioxi-
dant potential (TRAP) was measured by Rice-Evans and
Miller’s inhibition assay with a calibration curve for tro-
lox.23 Elimination of peroxyl radicals (ROO$) in plasma was
verified by Guohua Cao’s oxygen radical absorbance ca-
pacity (ORAC) method.24 The activities of antioxidant en-
zymes in red blood cells, including CAT, GSH-Px, and
SOD, were measured using an ultraviolet/visible light
spectrometer.25–27 Polymorphisms of the PPAR-c gene
C1431T (exon 6, rs3856806) were conducted by polymerase
chain reaction (PCR) restriction fragment length polymor-
phism with 50 -TAC CCT TCC TCC CCA CCT AT-3 and 50 -
GTG GCT CAG GAC TCT CTG CT-30 as the forward and
reverse primers, respectively.28 The extracted 100–200 ng of
template DNA (LaboPass Blood Mini Kit; Cosmo Gene-
tech, Seoul, Korea), 0.5 lM primers, Tris buffer (pH 8.3),
200 lM Taq DNA polymerase (Koma Biotech, Inc., Seoul,
Korea), and distilled water were used to amplify the target
template using a PCR machine (iCycler iQ PCR system;
Bio-Rad). Restriction enzymes were added to the above
complex and electrophoresis was performed in a 2% agarose
gel. The results were confirmed with an ultraviolet transil-
luminator after staining with EtBr. H447H gene variants
included CC (wild), CT (hetero), and TT (mutant).
Dietary intakes
At the initial visit and on the 4th and 12th weeks, nutrient
intakes using a three-day food record (including two con-
121
secutive days and one weekend day) with a 24-h recall were
analyzed using the CanPro3.0 software (The Korean Nu-
trition Society).
Statistical analysis
Statistical analyses were performed using SPSS 18.0 for
Windows (SPSS, Inc., Chicago, IL, USA). Data are pre-
sented as the mean – standard error (SE). Mean differences
before and after supplementation were tested by t-tests or
analysis of variance. Pearson’s correlation coefficients (r)
were used to describe the linear association between vari-
ables. Values of P < .05 were considered statistically sig-
nificant. Gene interactions and supplementation among the
test and placebo groups were evaluated by v2 tests, and
relationships among the factors and genes were evaluated by
paired t-test and analysis of variance after adjusting for age,
sex, and BMI.
RESULTS
Characteristics of the subjects
Table 2 shows the baseline characteristics of the subjects
with the C and T alleles of the PPAR-c polymorphism. The
relative frequencies (RF) of PPAR-c genotypes CC, TC, and
TT in 58 subjects were 70% (n = 41), 25.9% (15), and 3.4%
(2), whereas those of PPAR-c alleles C and T were 81.6%
and 18.4%, respectively. RF of the PPAR-c mutant T allele
(0.16) was slightly lower than in Caucasians and even
Chinese (RF = 0.24). There were no significant differences
in baseline characteristics (anthropometric, lipid profiles,
glucose tolerance, hematology and antioxidant parameters)
among the four groups, with one exception. Those subjects
in the doenjang-supplemented group with the C allele had a
significantly lower initial serum GSH-Px activity compared
with the other three groups. Significantly higher VFA/SFA
and Na/K ratios were shown in T allele subjects of the
doenjang group compared with those in the others.
The correlation among the parameters
BMI was positively correlated only with the fat mass
(P < .001) after age and sex were adjusted. Positive corre-
lation between waist and WHR, BP and SFA, TC and apo B,
HDL and apo A-I, LDL and apoB-100, TG/HDL and
HbA1C, FFA and energy intake, insulin and HOMA-IR, and
DNA tail length and CAT activity were significant (P < .01).
On the other hand, negative correlations between TFA and
TRAP, FFA and Na excretion, ORAC and carbohydrate
intake, and fat and K intake were significant (P < .01; data
not shown).
Differences on anthropometry in doenjang intervention
according to PPAR-c polymorphism
Doenjang supplementation significantly decreased VFA
(P = .005) compared with the placebo treatment, even
though the total fat mass content did not differ when age,
sex, and BMI were adjusted (Table 3). Otherwise, VFA
 122 CHA ET AL.

Table 2. Baseline Characteristics Adjusted by Age, Sex, and Body Mass Index
According to C and T Alleles of PPAR-c Gene

C allele T allele

Placebo (n = 39) Test (n = 41) Placebo (n = 11) Test (n = 7) P-value

Anthropometrics
BMI kg/m2 26.8 – 2.1 26.9 – 2.3 26.0 – 1.0 27.4 – 2.1 NS
WC cm 90.1 – 0.5ab 92.0 – 0.4b** 87.4 – 1.4a 90.5 – 1.9ab 0.044
WHR 0.93 – 0.00 0.93 – 0.00 0.90 – 0.01 0.92 – 0.02 NS
SBP mmHg 116.0 – 1.0 120.7 – 1.0** 121.7 – 2.3 117.2 – 3.3 NS
DBP mmHg 73.8 – 0.7 77.3 – 0.7** 79.1 – 1.5 77.1 – 2.2 NS
Fat mass kg 22.5 – 0.2 22.3 – 0.2 20.5 – 0.3 20.8 – 0.4 NS
VFA mm2 6913.7 – 205.6 7086.1 – 203.1 7120.0 – 352.8 7785.0 – 494.3 NS
SFA mm2 27,421.2 – 1841.9 26,901.6 – 1819.7 20,889.5 – 985.1 20,667.6 – 1380.1 NS
VFA/SFA ratio 0.33 – 0.01a 0.31 – 0.01a 0.40 – 0.03ab 0.45 – 0.04b 0.041
Lipid profiles
TC mg/dL 193.3 – 2.9 189.5 – 2.9 174.4 – 6.8 179.6 – 9.5 NS
TG mg/dL 99.0 – 3.0 90.3 – 3.0* 79.7 – 8.6 92.2 – 12.1 NS
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HDL mg/dL 53.5 – 1.0 54.6 – 1.0 58.4 – 1.8 50.7 – 2.5 NS
LDL mg/dL 130.8 – 2.8 121.8 – 2.7* 105.8 – 7.4 117.4 – 10.4 NS
TG/HDL 1.9 – 0.1 1.7 – 0.1 1.5 – 0.2 1.8 – 0.2 NS
FFA lEq/L 601.9 – 20.5 572.5 – 20.3 658.1 – 34.2 450.1 – 47.9** NS
Apo A1 mg/dL 1.42 – 0.02 1.48 – 0.02 1.54 – 0.04 1.42 – 0.06 NS
Apo B mg/dL 0.85 – 0.02 0.82 – 0.02 0.70 – 0.06 0.77 – 0.08 NS
Insulin resistance
Glucose mg/dL 83.9 – 0.9 81.9 – 0.9 87.5 – 1.9 87.2 – 2.6 NS
Insulin lU/mL 5.9 – 0.4 5.6 – 0.4 9.0 – 1.0 3.7 – 1.4* NS
HOMA-IR 1.3 – 0.1 1.1 – 0.1 2.0 – 0.3 0.8 – 0.4* NS
Fructosamin lM/L 247.8 – 2.2 244.2 – 2.2 258.3 – 6.9 244.5 – 9.7 NS
HbA1C % 5.40 – 0.04 5.44 – 0.04 5.46 – 0.11 5.31 – 0.15 NS
Tissue toxicity
AST IU/L 19.0 – 0.4 20.2 – 0.4* 21.6 – 0.8 20.0 – 1.1 NS
ALT IU/L 18.9 – 0.7 19.6 – 0.7 23.4 – 1.8 20.5 – 2.6 NS
Creatinine mg/dL 0.68 – 0.01 0.69 – 0.01 0.86 – 0.05 0.74 – 0.06 NS
Na excretion mmol/day 186.3 – 7.3 220.6 – 7.2** 171.3 – 13.6 188.8 – 19.1 NS
Antioxidant
DNA in tail % 20.8 – 0.3 21.5 – 0.3 19.5 – 0.7 22.6 – 1.0* NS
Tail length lm 39.9 – 0.3 38.9 – 0.3* 37.5 – 0.8 42.0 – 1.1 NS
ORAC TE/mL 19.2 – 0.3 19.9 – 0.2 19.6 – 0.9 19.2 – 1.0 NS
TRAP mM/L 1.723 – 0.002 1.716 – 0.002** 1.730 – 0.004 1.722 – 0.005 NS
Catalase K/gHb 34.4 – 1.2 31.5 – 1.2 33.9 – 2.9 44.1 – 4.1 NS
GSH-Px U/gHb 31.3 – 1.6b 25.1 – 1.6a** 36.5 – 2.6b 36.9 – 3.6b 0.011
SOD U/gHb 1886.6 – 25.4 1930.4 – 25.1 2142.4 – 76.5 1865.9 – 107.2 NS
Nutrition intake
Energy Kcal 1551.1 – 32.2 1446.2 – 32.6* 1492.6 – 87.6 1662.6 – 122.7 NS
Carbohydrate g/day 221.8 – 5.9 225.1 – 6.0 219.2 – 17.6 264.4 – 24.7 NS
Protein g/day 64.4 – 1.8 55.7 – 1.8** 57.1 – 4.3 60.9 – 6.0 NS
Fat g/day 43.6 – 1.4b 35.2 – 1.4a*** 39.0 – 2.4ab 37.2 – 3.4a 0.006
Sodium (Na) mg/day 3569.6 – 106.9 3669.5 – 108.2 3389.9 – 234.0 4016.2 – 327.8 NS
Potassium (K) mg/day 2353.6 – 58.6 2158.9 – 59.3* 2222.1 – 163.5 2108.5 – 229.0 NS
Na/K ratio 1.54 – 0.04a 1.71 – 0.04ab** 1.57 – 0.08a 1.93 – 0.12b* 0.032

Values are presented as mean – SE, adjusted by age (19–65 years), sex [male (n = 8), female (n = 43)], and BMI. P-values indicate significant differences among
the four groups.
abcd
Different superscript letters in the same row indicate statistically significant differences at P < .05.
Significant differences between the test and placebo groups are indicated with asterisks (*P < .05, **P < .01, ***P < .001) in the T and C alleles.
BMI, body mass index; WC, waist circumference; WHR, waist-to-hip ratio; SBP, systolic blood pressure; DBP, diastolic blood pressure; VFA, visceral fat area;
SFA, subcutaneous fat area; TC, total cholesterol; TG, triglycerides; HDL, high-density lipoprotein; LDL, low-density lipoprotein; FFA, free fatty acid; HOMA-IR,
homeostatic model assessment insulin resistance; HbA1C, hemoglobin A1C; AST, aspartate aminotransferase; ALT, alanine aminotransferase; ORAC, oxygen
radical absorbance capacity; TRAP, total radical-trapping antioxidant potential; NS, nonsignificance.
 INTERACTION OF PPAR-c AND FERMENTED SOYBEAN ON OBESITY 123

Table 3. Effects of Doenjang Supplementation on the Age-, Sex-, and Body Mass Index–Adjusted Characteristics
According to PPAR-c Polymorphism

C wild allele T mutant allele

PPAR-c Placebo (n = 39) Test (n = 41) Placebo (n = 11) Test (n = 7) P-value

Anthropometrics
WHR - 0.04 – 0.01 - 0.02 – 0.01 0.002 – 0.01 0.01 – 0.02 NS
SBP mmHg - 2.6 – 1.0a 2.1 – 1.0b** 3.3 – 2.7b 5.3 – 3.6b 0.034
DBP mmHg - 0.6 – 0.7 1.9 – 0.7* - 3.2 – 1.8 5.1 – 2.4* NS
Fat mass kg - 0.5 – 0.1 - 0.5 – 0.1 - 0.2 – 0.1 - 0.6 – 0.2 NS
Lean mass kg - 0.05 – 0.06 - 0.03 – 0.06 0.08 – 0.10 0.33 – 0.14 NS
VFA cm2 - 42.3 – 147.9c - 753.7 – 146.1b** - 338.0 – 161.8b - 1766.0 – 217.9a*** 0.005
SFA cm2 - 8419.4 – 1642.4 - 7848.1 – 1622.5 - 2864.1 – 722.9 - 5938.8 – 973.5* NS
Lipid profiles
TC mg/dL - 4.5 – 1.5 - 11.3 – 1.5** - 8.3 – 2.8 - 10.5 – 3.8 NS
TG mg/dL 6.4 – 3.7 - 0.1 – 3.6 14.7 – 5.6 - 13.3 – 7.6x NS
HDL mg/dL - 2.5 – 0.7 - 0.3 – 0.7* - 3.9 – 1.4 - 2.4 – 1.9 NS
LDL mg/dL - 11.9 – 1.3 - 12.6 – 1.3 - 13.1 – 2.7 - 13.8 – 3.6 NS
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FFA lEq/L - 68.9 – 22.1a 88.1 – 21.9b - 54.9 – 36.1a 56.7 – 48.6ab 0.011
Apo A1 mg/dL - 0.17 – 0.01 - 0.19 – 0.01 - 0.25 – 0.03 - 0.22 – 0.04 NS
Apo B mg/dL - 0.14 – 0.01 - 0.16 – 0.01 - 0.12 – 0.03 - 0.15 – 0.03 NS
Insulin resistance
Glucose mg/dL - 4.2 – 0.8 - 1.8 – 0.8* - 7.1 – 1.5 - 7.2 – 2.0 NS
Insulin lU/mL - 1.1 – 0.4a 1.3 – 0.4b - 1.9 – 0.9a 0.4 – 1.2b 0.018
HOMA-IR - 0.3 – 0.1a 0.3 – 0.1b*** - 0.5 – 0.2a 0.1 – 0.3b 0.009
Fructosamin lM/L - 6.3 – 1.4 - 10.0 – 1.4 - 8.7 – 2.1 - 15.7 – 2.8 NS
HbA1C % - 0.16 – 0.03 - 0.90 – 0.03 - 0.25 – 0.07 - 0.28 – 0.10 NS
Hematology
WBC · 103 cells/lL - 0.75 – 0.17 - 0.17 – 0.17* - 0.37 – 0.23 - 0.67 – 0.29 NS
RBC · 103 cells/lL - 0.10 – 0.03 - 0.17 – 0.03 - 0.08 – 0.05 - 0.13 – 0.07 NS
Hb g/dL - 0.3 – 0.1 - 0.4 – 0.1 - 0.4 – 0.2 - 0.4 – 0.2 NS
Hct % - 0.8 – 0.2 - 1.4 – 0.2 - 1.1 – 0.4 - 1.0 – 0.5 NS
Platelet · 103 cells/lL - 6.4 – 6.7 - 9.9 – 6.5 - 2.8 – 6.7 - 10.4 – 8.8 NS
Tissue toxicity
AST IU/L - 0.50 – 0.40 - 0.95 – 0.39 - 2.9 – 0.56 - 1.30 – 0.75 NS
ALT IU/L - 0.9 – 0.70 0.6 – 0.70 - 5.2 – 1.20 - 0.4 – 1.70* NS
BUN mg/dL - 0.84 – 0.37 - 0.34 – 0.36 - 0.96 – 0.55 - 0.08 – 0.74 NS
Creatinine mg/dL 0.07 – 0.02 0.02 – 0.02* 0.05 – 0.05 0.02 – 0.07 NS
Na excretion mmol/day 3.1 – 15.1ab - 8.9 – 14.7a 105.6 – 23.5b - 5.6 – 30.3a* 0.014
Antioxidants
Tail DNA 4.4 – 1.1 3.7 – 1.1 2.9 – 1.3 0.9 – 1.7 NS
Tail length 3.1 – 0.8 1.1 – 0.7 3.2 – 1.1 - 2.6 – 1.4* NS
ORAC - 0.7 – 0.4a 1.4 – 0.4b** 2.5 – 1.4c 1.0 – 1.6b 0.002
TRAP - 0.09 – 0.00 - 0.08 – 0.00 - 0.10 – 0.01 - 0.09 – 0.01 NS
Catalase - 14.9 – 2.5ab - 9.4 – 2.4b - 17.4 – 4.0ab - 29.2 – 5.2a 0.021
GSH-Px - 1.9 – 2.0 1.0 – 2.0 - 7.3 – 3.2 - 8.6 – 4.2 NS
SOD 217.5 – 42.6* 65.4 – 41.5* 222.5 – 120.0 77.4 – 155.0 NS
Nutrition intakes
Energy kcal - 56.4 – 48.5 51.1 – 48.5 - 126.5 – 84.7 - 350.3 – 109.5 NS
Carbohydrate g/day 120.5 – 6.4b - 12.5 – 6.4a* - 9.2 – 7.8ab - 32.8 – 10.1a 0.009
Protein g/day - 4.1 – 2.3 0.1 – 2.3 1.0 – 5.2 5.0 – 6.7 NS
Fat g/day - 6.1 – 2.4a - 2.7 – 2.4ab 4.6 – 3.8b 5.4 – 4.9b 0.042
Sodium (Na) mg/day 55.4 – 164.3b - 318.9 – 164.3a 68.3 – 273.4b - 421.7 – 353.3a 0.016
Potassium (K) mg/day - 212.3 – 76.3 - 92.7 – 76.3 - 415.1 – 162.7 224.4 – 210.3* NS
Na/K ratio 0.2 – 0.1b - 0.1 – 0.1a** 0.4 – 0.1b - 0.4 – 0.1a** 0.001

Values are presented as mean – SE, adjusted by age (19–65 years), sex [male (n = 8), female (n = 43)], and BMI. P-values indicate significant differences among
the four groups.
abcd
Different superscript letters in the same row indicate statistically significant differences at P < .05.
Significant differences between the test and placebo groups are indicated with asterisks (*P < .05, **P < .01, and ***P < .001).
WBC, whole blood cell count; RBC, red blood cell count; Hb, hemoglobin; Hct, hematocrit; BUN, blood urea nitrogen.
 124 CHA ET AL.
Downloaded by University Of Melbourne from www.liebertpub.com at 04/30/18. For personal use only.

FIG. 1. Presentation of visceral fat area and subcutaneous fat area regression according to increasing age in subjects having PPAR-c poly-
morphisms such as C wild and T mutant allele. B, wild + test (——); -, mutant + test (—); +, wild + placebo (); :, mutant + placebo (––
—). Color images available online at www.liebertpub.com/jmf

reduction was increased in those with the PPAR-c T allele
rather than the C allele. VFA was increased with increasing
age, in particular, > 40 years of age. We found that the PPAR-c
gene was positively correlated with VFA accumulation and
aging, but mutant of PPAR-c decreased SFA. (Fig. 1)
Differences in blood chemistry in doenjang intervention
according to PPAR-c polymorphism
Plasma FFA, insulin, and HOMA-IR levels were sig-
nificantly increased with increased catabolism of VFA
upon doenjang treatment in those with both alleles com-
pared with placebo. The Na excretion was higher in the
placebo group compared with doenjang group, to a level
corresponding to that of decreased systolic BP, and Na was
excreted 30-fold higher in those with the T allele than the C
allele. ORAC was increased in those with the wild-type
allele, but was decreased in those with either the mutant
allele or after doenjang pills were administered for 12
weeks. CAT activity was increased twofold in the doen-
jang-treated group with the C allele, but this phenomenon
was reversed in those with the T allele. However, SOD was
decreased by doenjang supplementation in the C allele
group, but there was no significant difference observed in
the T allele group. GSH did not differ after age, sex, and
BMI were adjusted.
Differences in dietary intakes due to doenjang intervention
according to PPAR-c polymorphism
We observed an interaction between doenjang and
PPAR-c in dietary carbohydrate and Na intakes. The reason
 INTERACTION OF PPAR-c AND FERMENTED SOYBEAN ON OBESITY
was that subjects with the mutant T allele who took doen-
jang consumed less dietary carbohydrates and sodium
compared with the C allele, even though there was no dif-
ference in the dietary doenjang intake. However, dietary fat
was slightly increased in the C allele group upon doenjang
supplementation. The dietary sodium intake was signifi-
cantly different when age, sex, and BMI were adjusted.
DISCUSSION
To best of our knowledge, this is the first study to report
antiobesity effects of long-term fermented soybean product,
Korean doenjang, in subjects with PPAR-c polymorphisms.
Doenjang significantly decreased VFA, but not SFA, by
increasing lipolysis in subjects with the mutant T allele.
Since the food intake and physical activity were not con-
trolled, those results of doenjang supplementation in over-
Downloaded by University Of Melbourne from www.liebertpub.com at 04/30/18. For personal use only.
weight subjects were very important to be generalized for
people following a regular diet and exercise regimen. Stu-
dies in rat adipocytes revealed that the soy isoflavonoids,
genistein and daidzein, enhance basal lipolysis, with
mechanisms possibly related to the regulation of fatty acid
metabolism using the nuclear receptors, PPAR-a and PPAR-
c.29,30 In an in vitro-based study, the authors demonstrated
that soy isoflavones doubled PPAR-a–directed gene ex-
pression, whereas they increased PPAR-c–directed gene
expression by 200–400%, resulting in an improved lipid and
glucose metabolism.31
Fermentation of soybeans alters compounds such as iso-
flavonoids and small peptides. As a result, isoflavonoid
aglycones, such as genistein and daidzein, which are more
active compounds representing 50–55% and 40–45% of
total isoflavones, respectively, are increased by more than
10- to 100-fold.9 Urea cycle intermediates (i.e., arginine, an
NO precursor, citrulline, and ornithine) are increased by 3.3-
to 33-fold depending on the length of fermentation.10 Small
peptides, Glu-Tyr and Glu-Phe, which are related to the
unique taste characteristics produced by soybean fermen-
tation, are involved in the prevention of hypertension.10
Isoflavones in foods and dietary supplements have both
advocates and critics in the health industry. The latter are
due to concerns about the estrogenic effects of isoflavones in
certain species. In this study, the subjects took 40 g of
doenjang a day, which is equivalent to 20–30 mg of iso-
flavones. This level produces plasma concentrations of
isoflavones ranging around 200–300 nM, which is typical of
many orally ingested therapeutics, but even daily doses of
1000 mg of soy isoflavones do not produce any significant
adverse effects.32 At low genistein levels, an estrogen-like
effect was observed, whereas at micromolar levels, PPAR-c
activation predominated.33 However, in a previous study, a
high dose of 160 mg of isoflavones (20 g of soy protein) over
12 weeks did not improve inflammatory parameters in an-
drogen-deprived men with prostate cancer.34
Genistein inhibits the proliferation and differentiation of
3T3-L1 cells, a preadipocyte cell line, by increasing the
CCAAT/enhancer-binding protein beta (C/EBP beta) ac-
tivity and impacting PPAR-c protein expression.35 Soybean
125
fermented for longer periods improves the insulin resis-
tance. PPAR-c acts as a central regulator of insulin sensi-
tivity by increasing insulin-stimulated IRS2-mediated
glucose uptake and TG accumulation.36 In another study,
genistein, daidzein, and small peptides ( < 3 kDa), which
were produced by longer fermentation of soybeans, in-
hibited preadipocyte differentiation and body weight gains
in vivo through PPAR-c activation.24 The mechanism of TG
accumulation in adipocytes of animals and humans is also
connected with PPAR-c by the mechanism of insulin sen-
sitivity. PPAR-c agonists, such as thiazolidinediones, have a
site-specific effect on the differentiation of human pre-
adipocytes, in particular, subcutaneous fat, although pre-
adipocytes from visceral fat are refractory to drugs like
troglitazone. Visceral fat accumulation is closely associated
with insulin-resistance syndromes, including impaired glu-
cose tolerance, hyperlipidemia, hypertension, and cardio-
vascular disease.37
We expected that the antioxidant effect of isoflavones in
fermented doenjang would mediate their cytoprotective ef-
fects even at low levels. Like other polyphenols, many
studies have shown that isoflavones can scavenge various
reactive oxygen species that are formed endogenously dur-
ing the innate immune response.38 In our results, ORAC was
increased in the C allele but was decreased in the T allele
after doenjang pills were administered for 12 weeks. CAT
activity was increased twofold in the doenjang-treated
group with the C allele, but this phenomenon was reversed
in those with the T allele. Contrary to the above results, SOD
was increased by placebo. One of the reasons was that some
ingredients in the placebo pills, such as cowpea, wheat, rice,
cocoa, and chrysanthemum, may be antioxidants, which
may influence the results. The decreased proinflammatory
responses, antiatherosclerotic activities, and antioxidant
effects were proposed as mechanisms underlying the bene-
ficial effect of PPAR-c activators. PPAR-c promotes ex-
pression of numerous gene products whose profile of
activity could be linked to oxidative stress suppression,
proper mitochondrial function, or lipid and glucose me-
tabolism and transport in non-neuronal tissues.39 More-
over, isoflavones prevent atherosclerosis by inhibiting
LDL oxidation and affecting signaling pathways, such as
activation of estrogen receptor beta (ERb) and PPARs.40
For example, PPAR-c ligands inhibit the cytokine-depen-
dent expression of adhesion molecules in the endotheli-
um.41 We found one interesting result: doenjang-
administered subjects with the mutant T allele consumed
less sodium compared with the C allele, an interesting
observation since soy improves the BP of spontaneously
hypertensive rats on a high-salt diet.42
The limitations of this study include the following: (1) the
number of subjects was not sufficient to identify the inter-
action between the PPAR-c gene (mutant T allele RF = 0.18)
and environment; therefore, we split the number of subjects
with polymorphisms into two allele groups and not three
genotypes; (2) nine subjects withdrew from the study—two
did not submit consent forms, two voluntarily withdrew for
personal reasons, and the remainder were excluded due to
 126
noncompliance with the study protocol. Even though our
study results may not be generalized to the entire Korean
population, our results on doenjang intervention can estab-
lish the basis for further functional food studies on obesity
prevention. However, the antiobesity effect of VFA in
overweight subjects with PPAR-c polymorphisms should be
confirmed in various populations, since genetic differences
among ethnic groups may be present. In conclusion, this
study suggests that doenjang has antiobesity and anti-
oxidative effects in overweight individuals with mutant al-
leles of PPAR-c.
ACKNOWLEDGMENTS
This study was funded by the Seoul City R&BD program
(10526) and the Ministry of Health & Welfare (A000385),
Republic of Korea. All authors participated in this work with
their substantive contributions. Y.-S.C. and S-.W.C. had the
Downloaded by University Of Melbourne from www.liebertpub.com at 04/30/18. For personal use only.
responsibility for conducting and performance of the RCT
projects. M.L. and K.P. participated in the biochemical
analysis and interpretation of data, and Y.K. and Y.P. car-
ried out the statistics.
AUTHOR DISCLOSURE STATEMENT
No competing financial interests exist.
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