Lactic-fermented egg white improves visceral fat obesity in Japanese subjects—double-blind, placebo-controlled study
Matsuoka et al. Lipids in Health and Disease
Lactic-fermented egg white improves visceral fat obesity in Japanese subjects-double-blind, placebo-controlled study
Ryosuke Matsuoka 2
Keiko Kamachi 1
Mika Usuda 2
Wei Wang 2
Yasunobu Masuda 2
Masaaki Kunou 2
Akira Tanaka 1
Kazunori Utsunomiya 0
0 Department of Internal Medicine, The Jikei University School of Medicine , 3-25-8, Nishi-shinbashi, Minato-ku, Tokyo 105-8461 , Japan
1 Nutrition Clinic, Kagawa Nutrition University , 3-24-3, Komagome, Toshima-ku, Tokyo 170-8481 , Japan
2 R&D Division, Kewpie Corporation , 2-5-7 Sengawa Kewport, Sengawa-Cho, Chofu-Shi, Tokyo 182-0002 , Japan
Background: It was reported that egg white protein (EWP) reduced body fat in rats. We developed a lacticfermented egg white (LE) that facilitates the consumption of egg whites by fermenting them with lactobacillus, and were able to study their intake in humans. In this double-blind, placebo-controlled design, we evaluated the effect of LE on visceral fat area (VFA). Methods: Participants included 37 adult males and females aged ≥40 years (VFA at navel ≥100 cm2). They were divided into two groups: the control group and the LE group. The control and LE groups consumed drinks containing whey and LE, respectively, for 12 weeks (providing 8 g protein/day). VFA was measured at baseline and at week 12 of intake. Abdominal girth was measured at baseline and at weeks 6 and 12. Results: LE intake decreased VFA significantly compared with baseline (−8.89 cm2, p < 0.05), and VFA was significantly lower than that in the control group (+1.71 cm2, p < 0.05). The LE group showed significant improvement in the ratio of visceral to subcutaneous fat area compared with baseline and the control group (p < 0.05). Conclusions: The results demonstrated that LE reduces VFA and improves the ratio of visceral to subcutaneous fat area. As other measurement items were not influenced, we concluded that LE improves visceral fat obesity. Trial registration: This clinical trial was retrospectively registered with the University hospital Medical Information Network (UMIN) Center, (UMIN000026949; registered on 11/04/2017; http://www.umin.ac.jp/).
Egg white; Visceral fat; Obesity; Japanese; Protein
Visceral fat obesity causes complications such as
dyslipidaemia, abnormal glucose tolerance, and hypertension.
Along with serum LDL-cholesterol concentration, it is a
risk factor for arteriosclerotic disease [
]. In Japan,
approximately 50% men and 20% women ≥40 years have
metabolic syndrome or pre-metabolic syndrome, which
is becoming more serious [
Hen eggs contain almost all essential nutrients (except
vitamin C and dietary fiber), and are known to be of
high nutritive value [
]. Attention has been drawn to the
cholesterol contained in eggs, and nutritional education
for dyslipidaemia recommends avoiding egg
consumption. Although the cholesterol in eggs is found in the
yolk, some reports have indicated that consumption of
egg yolk does not increase blood cholesterol levels [
or mortality due to coronary heart disease [
we considered whether the constituent that improves
lipid metabolism is contained in eggs, specifically egg
white protein (EWP). EWP is high in protein and low in
fat and has been reported to reduce serum total and
LDL cholesterol levels [
EWP is said to lower blood cholesterol by inhibiting
cholesterol absorption in the small intestines. However,
in doing so, triglyceride absorption is also inhibited [
Therefore, EWP not only reduces serum LDL cholesterol
level but is also believed to prevent arteriosclerotic
disease by reducing visceral fat.
In an experiment on rats, Matsuoka et al. reported
that egg white proteins could reduce body and
abdominal fat [
]. However, it is unclear whether the
reduction was effective in human.
The consumption of egg white itself poses a problem
in terms of taste and physical properties. We therefore
developed a lactic-fermented egg white (LE) that
facilitates the consumption of egg whites by fermenting them
with lactobacillus, and were able to study their intake in
]. In a previous study, it was reported
that the EWPs ovalbumin and ovotransferrin could
inhibit lipid absorption . It has been reported that
beta-conglycinin, a soy protein, reduced visceral fat at
the speed of 5 g/day [
]. Ovalbumin and ovotransferrin
are reported to contain 54% and 11% EWP, respectively
]. If we suppose that ovalbumin and ovotransferrin
have the same visceral fat-reducing effect as
betaconglycinin at a daily intake of 5 g, a daily intake of 8 g
of EWP would be required. We therefore conducted a
double-blind, placebo-controlled study to evaluate the
visceral fat-reducing effect of protein in Japanese adults
with visceral fat obesity, via daily intake of LE for
The control drink and the drink containing LE were
prepared by Co-op Foods Co., Ltd. (Tokyo, Japan). Flavors,
sweeteners and water were added to the whey
(NipponShinyaku Co., Ltd., Kyoto, Japan) and LE [
Egg Corporation, Tokyo, Japan) of each drink. Once
homogeneously dispersed, the preparation was
heatsterilized. Measurement using the Kjeldahl method [
indicated that both drinks contained 8 g of protein per
180 g serving. Ovalbumin content in LE was assessed
by the sandwich ELISA procedure using anti-chicken
ovalbumin polyclonal antibody, and horseradish
peroxidaselabeled anti-chicken ovalbumin polyclonal antibody,
respectively. A commercial kit, Egg (Ovalbumin) ELISA
kit II (Morinaga Institute of Biological Science Inc.,
Yokohama, Japan) was used with absorbance detected
at 450 nm by a multi-detection microplate reader
(Powerscan® HT, DS Pharma Biomedical Co. Ltd.,
Osaka, Japan). Ovalbumin content of the LE drink was
4.17 g/180 g, as measured via sandwich ELISA. Based on a
previous report that ovotransferrin accounts for 11%
of total EWP [
], the content of ovotransferrin was
calculated to be 0.88 g/180 g LE (Calculated value).
Participants and test methods
Recruitment was done per the following process:
Japanese subjects with body mass index (BMI) (≥25),
aged more than 40 years (male) or postmenopausal
female, who were not undergoing treatment for
hyperlipidemia or diabetes, who had no subjective
symptoms of gout and who were capable of filling
out the required forms, such as self-diagnosis forms,
and of visiting a designated institution as scheduled.
The exclusion criteria were as follows: taking drugs
that could potentially affect the test results (e.g.,
antihyperlipidemic agents, anti-diabetic agents, oral
corticosteroid formulations, and antihypertensive agents);
regular consumption of foods for specified health uses
that could potentially affect test results; excessive
alcohol consumption; suspected allergic reactions
(particularly to egg and milk); participation in other
clinical studies; a history of serious liver damage,
kidney damage, or myocardial infarction; a history of, or
current, hepatitis; and serious anemia. In this study,
80 adults were screened and 48 individuals with
visceral fat area (VFA) (≥100 cm2) were included as test
The study followed a double-blind, parallel-arm
design. Participants were divided into two groups: a
control group and LE group. The control group was given a
drink containing whey (8 g of protein), and the LE group
was given a drink containing LE (8 g of protein) every
day at breakfast for 12 continuous weeks.
At 0 and 12 weeks of intake, computed tomographic
(CT) scanning of the abdomen (Aquilion™, Toshiba
Medical Systems Corporation, Tochigi, Japan) was
performed, and VFA was measured at the navel (Fat Scan,
East Japan Institute of Technology Co., Ltd., Ibaraki,
Japan). CT scans were conducted at the Nakajima Clinic
(Tokyo, Japan). At weeks 0, 6, and 12, bodyweight, blood
pressure, and abdominal girth, respectively, were
measured after fasting overnight. Blood was sampled from a
forearm vein, and blood tests were performed at
baseline, and weeks 6 and 12 of intake.
Peripheral blood test was performed using flow
cytometry. Serum analysis included the following items: total
cholesterol (enzyme method); HDL cholesterol (direct
method); triglyceride (enzyme method); free fatty acid
(FFA; enzyme method); phospholipid (enzyme method);
glucose (hexokinase UV test); HbA1c (latex coagulating
method); RLP cholesterol (immunoadsorption); insulin
(CLEIA method); AST (JSCC transferable method); ALT
(JSCC transferable method); gamma-GTP (JSCC
transferable method); blood urea nitrogen (BUN; urease
LED UV method); creatinine (enzyme method); and uric
acid (enzyme method). LDL cholesterol concentration
was calculated using the Friedewald formula [
tests were performed by SRL Corporation (Tokyo,
The nutritional value of the content of meals recorded
by participants over a three-day period was calculated
using Excel Eiyoukun Food Frequency Questionnaire
version 5 (Kenpakusha, Tokyo, Japan). The 2010
Standard Table of Food Composition in Japan database was
used in this nutrition-calculation software.
Test results are shown as mean ± standard error.
Comparisons with baseline values were performed using
paired t-tests for VFA and Dunnett’s test for all other
items. Comparisons with the control group were
performed using the student’s t-test. A significance level of
less than 5% was considered substantial. Statistical
analyses were performed using SPSS version 20 (SPSS Inc.,
Tokyo, Japan). Change in VFA is indicated by Δ cm2.
Adherence and participant attributes
During the study period, two participants withdrew from
the study; one individual in the control group could not
get accustomed to the taste of the test food and one
individual in the LE group experienced melena. The
latter’s condition was assumed to be due to the
participant’s tendency toward constipation and the
melena was thus determined to not be causally related
to the test food (Fig. 1).
From the control group, seven participants were
excluded: two of whom did not fast prior to blood
sampling; four who did not complete the food diary
sufficiently to track actual food intake; and one who was
inconsistent in the timing of test food consumption.
From the LE group, two participants were excluded: one
who did not fast prior to blood sampling and one who
completed the food diary insufficiently (Fig.1). From the
above, we analyzed 18 individuals in the control group
and 19 in the LE group. There were no significant
differences observed at baseline between the two groups in
terms of age, height, or BMI (p > 0.05, Table 1).
There was no significant change in the amount of food
consumed during the study period in either group
The abdominal girth of the control group was
significantly lower at 12 weeks than at baseline. No significant
difference was observed for any other items between the
control and LE groups, nor compared to baseline
Fat area at the navel
There was no significant difference in the total fat area
in the LE group compared to baseline or the control
group (Table 3). Although no significant difference was
Fig. 1 Participant flow through a randomized trial
observed between the two groups in terms of VFA and
subcutaneous fat area, the LE group showed significantly
less VFA, and a significant increase in subcutaneous fat
area compared to baseline (p < 0.05). The control group
showed no significant difference compared to baseline
Calculation of the visceral to subcutaneous fat ratio
indicated a significantly lower ratio in the LE group
compared to baseline. There was no significant difference in
the control group compared to baseline or between the
two groups (Table 3).
Calculation of the change in VFA revealed a significant
reduction in the LE group compared to both baseline
and the control group. With regards to the change in
visceral to subcutaneous fat ratio at the navel (Δ), the
value was significantly reduced by the intake of LE
compared to the control group and baseline ratio (Fig. 2 and
Serum biochemical analysis
There was no significant variation observed in serum
total cholesterol, LDL cholesterol, RLP cholesterol,
triglycerides, phospholipids, insulin, or blood glucose
levels. While the LE group exhibited significantly lower
FFA concentrations compared to the control group in
week 6 (p < 0.05), there was no significant difference
observed in week 12. Moreover, while the LE group
showed significantly lower HDL-cholesterol levels in
week 6 compared to baseline (p < 0.05), there was no
significant difference observed in week 12 (Table 4).
The control group exhibited significantly high mean
corpuscular volume (MCV) in week 6 compared to
baseline, whereas mean corpuscular hemoglobin level
(MCHC) in weeks 6 and 12 were significantly lower than
baseline (p < 0.05). The LE group showed significantly
higher MCV in week 6 compared to baseline (p < 0.05;
Table 5). Throughout the entire study period, MCV and
MCHC were significantly lower in the LE group
compared to the control group (p < 0.05). All items’ values
stayed within the normal range and no particular
problems were observed (Table 5).
Liver and kidney functions indices
While the LE group exhibited significantly higher BUN
levels in week 6 compared to baseline (p < 0.05), BUN
was within the normal range, with no significant
difference observed in week 12. The LE group showed
significantly lower creatinine levels than the control group at
baseline and in week 12 (p < 0.05). However, there was
no significant variation observed throughout the intake
period, and because changes were within the normal
range, this was not considered to pose any problem. For
all other items, no influence of the test foods was
observed (Table 5).
Results of this study indicated that when 8 g of LE (i.e.,
EWP) was consumed daily for 12 weeks, VFA was
significantly reduced compared to baseline and the control
group. Therefore, we concluded that LE has a visceral
fat-reduction effect. It has been previously reported that
the visceral fat-reduction food constituents include
] and polyphenol [
]. The degree by which
these constituents reduced visceral fat of the food
consumed for 12 weeks was −10.3 cm2 for catechine and
−7.9 cm2 for polyphenol, compared to a reduction of
8.89 cm2 by LE in this study. This finding suggests that
LE has the same effect as food constituents that are
generally considered to reduce visceral fat. It has been
reported that protein-derived constituents also reduce
visceral fat, and therefore, in this study we evaluated the
effect of a daily intake of 8 g of lactic acid-fermented
albumin as protein, based on a report on beta-conglycinin
in soybeans [
]. However, this study did not
Mean ± SE of 18 (control) and 19 (LE)
Control Control group, LE Lactic fermented egg white group
examine whether less than 8 g of EWP would exhibit a
visceral fat-reduction effect and this requires further
study. Furthermore, it has been reported that lactoferrin
(LF), found in milk protein, has a visceral fat-reduction
effect . It has been found that a daily intake of
300 mg of LF significantly reduced VFA compared to
baseline and the control group. In this study, the control
group was given whey, which contains LF. It was
expected that a daily intake of 8 g of whey would
correspond to a daily intake of 450 mg of LF and was
therefore expected to have a visceral fat-reduction effect.
However, whey did not reduce visceral fat in the control
group. A possible explanation is that whey contains
many nutrients other than LF, which may have
competed. It has previously been reported that a daily
intake of 28 g of whey had no effect on visceral fat [
It was reported that it needs to be subjected to enteric
coating to exert its visceral fat–reducing effects because
LF itself is vulnerable to heating and is easily degraded
by gastric acid [
]. This should explain why the whey
sample used in the present study failed to reduce visceral
Furthermore, in this study the ratio of visceral to
subcutaneous fat was significantly reduced after
consumption of LE compared to baseline ratio and the control
group. The visceral to subcutaneous fat ratio is an
indicator of visceral fat obesity, with a ratio > 0.4 indicating
visceral fat obesity [
]. In our study, LE not only
reduced VFA but also considerably reduced the visceral
to subcutaneous fat ratio; therefore, we concluded that
the symptoms of visceral fat obesity had reduced.
Although it is unclear why LE intake reduces visceral
fat, it has been reported that EWP, the main constituent
of LE, has an inhibitory effect on the absorption of
]. EWP inhibited the absorption of fat,
thereby functioning to reduce lipid content within bile
acid micelles. The components involved in this process
are ovalbumin and ovotransferrin, both of which reduce
lipid content within bile acid micelles to an equivalent
extent. No other components have been shown to affect
this process [
]. As such, we inferred that these two
components might also have come into play in this
experiment. Especially, the underlying cause of this has
been reported to involve lipase inhibitory activity in
], the main constituent of EWP, as well as
the inhibitory effect of lipid micelles [
Furthermore, ovalbumin has been found to form FFAs and
]. Therefore, it is believed that EWP can
inhibit the absorption of lipids by way of its
EWP has an amino acid score of 100, with a higher
rate of net protein utilization than soy and milk proteins
]. In a previous examination of the effect of EWP on
carcass protein contents in rat compared to casein, we
reported that EWP showed significantly higher levels
than casein [
]. As the main constituent of muscle is
protein, it is believed that the increase in carcass protein
contents in rats were due to increased muscle mass. In
general, however, a muscle does not increase without
exercise; thus, further studies should be conducted to
determine whether a muscle increases because of the high
net protein utilization and increased body protein. The
fact that muscle burns fat in the event of increased
muscle may be one explanation for the observed
reduction in visceral fat.
It is generally believed that active food constituents
that reduce visceral fat are often those whose structures
do not change through absorption or metabolic process
(e.g., polyphenol), the effects of which are easily
evaluated in vivo. In contrast, proteins become effective after
being broken down into peptides and amino acids in the
digestive tract. With regards to the details of the active
constituents contained in EWP, we believe that the
visceral fat-reduction mechanism should first be clarified
and then confirmed via in vitro evaluation.
In this study, participants consumed either LE or whey
drink at breakfast. Suzuki et al. reported that
preexercise intake of EWP increases muscle and muscular
]. It has been reported that consuming hen’s
eggs at breakfast induced satiety and reduced
participants’ body fat [
]. However, in this study, while
participants’ energy intake hardly changed and the lactic acid
drink did not affect their satiety, we confirmed that
visceral fat was reduced. On the contrary, protein intake
following muscle strength training is recommended to
increase muscles [
]. Therefore, we believe that
consuming LE after dinner or before sleeping might reduce
visceral fat more effectively. Furthermore, if the visceral
fat-reduction mechanism of EWP lies in the inhibition
of lipid absorption [
], it may be preferable to consume
LE at dinnertime, the meal with the highest energy
intake. Moreover, previous research has reported that the
circadian rhythm gene BMAL1, which is involved in
lipid accumulation [
], is abundantly secreted at night.
Therefore, we believe that the consumption of LE at
dinnertime could more effectively inhibit the accumulation
of lipids. Per these factors, while the present study
indicated that LE intake at breakfast reduces VFA, the
timing at which it effectively reduces visceral fat was not
clarified and should be examined in future studies.
In the present study, the blood biomarkers exhibited
no significant differences between week 0, week 6, and
week 12. Although the present study involved subjects
with slightly high levels of visceral fat weight, the other
parameters such as blood lipids were within the normal
range. This might have resulted in no significant changes
being observed in the blood biomarkers. In previous
studies, subjects with slightly high levels of serum
cholesterol showed significant decreases in serum total
cholesterol and LDL-cholesterol concentrations at week 4
and week 8 during 8 weeks of LE consumption as
compared with prior to consumption [
]. This suggests the
possibility that marked improvements may be achieved
if subjects with borderline or abnormal blood biomarker
values are involved.
Concern has been raised that EWP intake could cause
allergies and biotin deficiency. In this study, individuals
with food allergies were excluded during recruitment. It
has been demonstrated that avidin found in raw egg
whites binds to biotin, thereby reducing its absorption
]. While the participants of this study were asked to
consume 8 g of EWP per day for 12 consecutive weeks,
there was no sign of clinical symptoms. A study in which
participants consumed 200 g of dried egg white
(approximately 160 g of EWP) daily reported signs of biotin
deficiency after week 7 [
]. The consumption period
was 12 weeks; however, the amount of EWP intake was
8 g, which was 1/20th of the amount of EWP that
caused biotin deficiency in the previous study, and may
explain the observed lack of biotin deficiency.
Furthermore, when egg white is in its raw state or heated, it has
been reported that electrophoretic avidine bands
disappear upon activation of pepsin . During the
digestive process, as pepsin is usually activated, we believe that
a daily intake of 8 g of LE for 12 weeks should not cause
biotin deficiency. Finally, we measured markers of liver
and kidney function. No abnormal increases or
decreases in numeric values were observed, suggesting that
EWP can be consumed safely.
Metabolic syndrome, triggered by obesity, leads to
hypertension, abnormal glucose tolerance, and is
ultimately considered a cause of arteriosclerotic diseases. If
we pursue this study further and clarify the visceral
fatreduction effect of EWP, we believe that it could
improve the quality of life of patients with obesity.
Furthermore, if the practical application of LE, which
facilitates the egg white intake, becomes available, we believe
that it would provide a good source of protein while
reducing visceral fat and might be utilized to promote the
health among the Japanese.
The results demonstrated that LE reduces VFA and
improves the ratio of visceral to subcutaneous fat area. As
other measurement items were not influenced, we
concluded that LE improves visceral fat obesity.
BMI: Body Mass Index; CT: Computed tomographic; EWP: Egg white protein;
LE: Lactic-fermented egg white; VFA: Visceral fat area
This study was supported completely by a grant received from Kewpie
Corporation, Japan. We thank Prof. Michihiro Sugano (Kyushu University, and
Prefectural University of Kumamoto) for providing technical advice about this
trial. The authors would like to thank Enago (www.enago.jp) for the English
Experimental work in Nutrition Clinic, Kagawa Nutrition University was
supported by Kewpie Corporation (Tokyo, Japan).
Availability of data and materials
The dataset supporting the conclusions of this article is included within the
RM, KK, MU, WW, YM, MK, AT, and KU conceived the study concept and
design. AT, as the principal investigator, was responsible for study logistics,
data acquisition and RM and KK for manuscript preparation. KK, MU, and WW
were responsible for conducting the trial, data collection, and performing
laboratory analysis. RM and AT carried out the statistical analysis. YM, MK,
and KU supervised the study design and commented on the manuscript. All
authors contributed to the intellectual content of the manuscript. All authors
read and approved the final amnuscript.
Ethics approval and consent to participate
This study was conducted in accordance with the Declaration of Helsinki,
and with the approval of the Ethics Committee of Kagawa Education
Institute of Nutrition (authorization no. 243; 13th March, 2013). Participants
received a thorough explanation of the study protocol and those who
provided consent were included. The study was conducted at the Nutritional
Clinic of Kagawa Education Institute of Nutrition.
Consent for publication
KK, AT, KU declare no conflict of interest. RM, MU, WW, Y.M. and M.K. are
employees of Kewpie Corporation. There are no other patents, products in
development or marketed products to declare.
Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.
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