N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX

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Yellow pea fiber improves glycemia and reduces

Clostridium leptum in diet-induced obese rats☆

Amanda J. Eslinger a , Lindsay K. Eller a, b , Raylene A. Reimer a, b,⁎

a
Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Calgary, Calgary, AB, Canada
b
Faculty of Kinesiology, University of Calgary, Calgary, AB, Canada

ARTI CLE I NFO A BS TRACT

Article history: Numerous studies have demonstrated the impact of functional fibers on gut microbiota and
Received 3 April 2014 metabolic health, but some less well-studied fibers and/or fractions of foods known to be
Revised 22 July 2014 high in fiber still warrant examination. The aim of this study was to assess the effect of
Accepted 25 July 2014 yellow pea-derived fractions varying in fiber and protein content on metabolic parameters
and gut microbiota in diet-induced obese rats. We hypothesized that the yellow pea fiber
Keywords: (PF) fraction would improve glycemia and alter gut microbiota. Rats were randomized to 1 of
Pulse grain 5 isoenergetic dietary treatments for 6 weeks: (1) control; (2) oligofructose (OFS); (3) yellow
Obesity PF; (4) yellow pea flour (PFL); or (5) yellow pea starch (PS). Glycemia, plasma gut hormones,
Glycemia body composition, hepatic triglyceride content, gut microbiota, and messenger RNA
Microbiome expression of genes related to hepatic fat metabolism were examined. Pea flour
Adiposity attenuated weight gain compared with control, PF, and PS (P < .05). Pea flour, PS, and OFS
Rat had significantly lower final percent body fat compared with control. Oligofructose but not
the pea fraction diets reduced food intake compared with control (P < .05). Pea fiber resulted
in lower fasting glucose and glucose area under the curve compared with control. Changes
in gut microbiota were fraction specific and included a decrease in Firmicutes (percent) for
OFS, PF, and PFL compared with control (P < .05). The Firmicutes/Bacteroidetes ratio was
reduced with OFS, PF, and PFL when compared with PS (P < .05). Taken together, this work
suggests that yellow pea-derived fractions are able to distinctly modulate metabolic
parameters and gut microbiota in obese rats.
© 2014 Elsevier Inc. All rights reserved.

Abbreviations: ACC, acetyl coA carboxylase; AUC, area under the curve; DIO, diet-induced obese; FAS, fatty acid synthase; GLP-1,
glucagon-like peptide 1; HFHS, high-fat/high-sucrose; mRNA, messenger RNA; MTT, meal tolerance test; OFS, oligofructose; OGTT, oral
glucose tolerance test; PF, pea fiber; PFL, pea flour; PS, pea starch; PYY, peptide tyrosine tyrosine; rRNA, ribosomal RNA; SCFA, short chain
fatty acid; SREBP-1c, sterol regulatory element-binding protein 1c.
☆
Supported in part by a research grant from The Agriculture Funding Consortium (Alberta Pulse Growers Commission, Agriculture and
Food Council of Alberta, and Minister of Advanced Education and Technology; grant no. 2008F030R). LKE was supported by a Natural
Sciences and Engineering Council of Canada Doctoral Scholarship and Canadian Diabetes Association Graduate Scholarship (DS-2-07-
2195-LE). LKE and AE declare no conflict of interest. RAR previously held a research grant from Beneo-Orafti Inc (Mannheim, Germany),
manufacturer of Raftilose P95, for a project unrelated to the current work.
⁎ Corresponding author at: Faculties of Kinesiology and Medicine, University of Calgary, 2500 University Drive NW, Calgary, AB T2N 1N4,
Canada. Tel.: +1 403 220 8218; fax: +1 403 284 3553.
E-mail address: reimer@ucalgary.ca (R.A. Reimer).

http://dx.doi.org/10.1016/j.nutres.2014.07.016
0271-5317/© 2014 Elsevier Inc. All rights reserved.

Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
2 N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX
1. Introduction
Obesity and its related comorbidities are increasing at an
unprecedented rate [1]. Parallel to this, there has been a
dramatic change in the human food supply. Many developed
regions, including Europe and North America, are character-
ized by a marked increase in processed food consumption and
a resultant decline in dietary fiber intake [2]. The benefits of
regular fiber consumption are numerous including decreased
risk of cardiovascular disease, improved glycemic control,
decreased risk of some cancers, and regulation of appetite and
body weight [3].
Gut hormones including glucagon-like peptide 1 (GLP-1) and
peptide tyrosine tyrosine (PYY) are involved in food intake and
energy balance [4,5]. The production and/or activity of these
hormones is often disrupted in obesity, which may contribute to
increased food intake and a change in energy balance [4,5].
Consumption of certain dietary fibers, including the prebiotic
fiber oligofructose (OFS), increases the secretion of GLP-1 and
PYY [6,7]. The endogenous release of GLP-1 and PYY—through
changes in food intake—may be part of the reason why some
dietary fibers improve physiologic health [8,9].
Furthermore, recent evidence points to a distinct relation-
ship between the composition of the intestinal microbiota and
obesity [10]. Although not unanimously shown [10-12], on the
one hand, obesity appears to be characterized by a decrease in
Bacteroidetes and a corresponding increase in Firmicutes,
which has been associated with increased energy extraction
during digestion [13]. On the other hand, certain strains of
bifidobacteria have been linked with improvements in met-
abolic health, including improved glucose tolerance, reduced
serum cholesterol and triglycerides, and lower serum levels of
leptin and interleukin 6 [14,15]. Although prebiotics are known
to increase bifidobacteria and reduce Firmicutes [16,17],
examination of the effects of other nondigestible carbohy-
drates, such as resistant starches and their fermentation end
products (ie, short chain fatty acids [SCFAs]), are increasingly
being examined for their effects on gut microbiota [18].
Pulses are a type of legume that are harvested for their dry
seed and include dry beans, lentils, chickpeas, and dry peas.
Pulse-based foods have the potential to alter satiety and the
gut microbiota due to their protein and carbohydrate (starch
and fiber) content [19]. Human and rodent studies as well as
epidemiological evidence suggest improvements in metabolic
health with the consumption of various pulses [19-27]. Recent
studies have shown that consumption of whole pulses for
8 weeks reduced symptoms of the metabolic syndrome in
overweight and obese adults [27]. In an attempt to identify the
active fraction of pulses, several studies have examined the
effects of the protein and fiber fractions on metabolic
outcomes. Smith et al [26] showed that healthy adults
consuming a single test meal of isolated yellow pea protein
but not isolated pea fiber (PF) had improved satiety and
glycemic response. The authors conclude that the protein but
not the fiber fraction of peas is responsible for the immediate
acute effects of yellow pea consumption, but long term
adaptation was not examined [26]. In overweight adults,
both whole and fractionated yellow pea flour (PFL) reduced
fasting insulin and insulin resistance, whereas whole yellow
Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
PFL also reduced android:gynoid fat ratio in women [25].
Chronic intake of fractionated but not whole yellow PFL was
shown to reduce postprandial energy expenditure in over-
weight adults [24]. In Golden Syrian hamsters, whole and
fractionated yellow PFL reduced insulin, and fractionated PFL
also decreased glucose levels compared with control [22]. The
whole yellow PFL was associated with a significant shift in
microbial composition with predominant changes in the
Firmicutes phylum [22]. Moreover, a randomized controlled
trial demonstrated that healthy adults consuming raffinose, a
chickpea oligosaccharide, had increased bifidobacteria [28];
similarly, rats fed pea and chickpea-containing diets had
greater bifidobacteria abundance [20]. Based on previous
work, it is clear that there are common but also some unique
physiologic effects attributable to the various fractions of
pulses, the most notably examined to date being yellow peas.
Therefore, the primary objective of this study was to
examine the effect of yellow pea-derived fractions on gut
hormone and glucose response and gut microbiota in a diet-
induced obese (DIO) rat model. A secondary objective was to
examine the effects of the fractions on body composition and
genes involved in hepatic lipid metabolism. It was hypothe-
sized that the yellow PF fraction would improve glycemia and
alter gut microbiota. Examination of 2 other yellow pea-
derived fractions (yellow PFL and yellow pea starch [PS] richer
in protein and starch, respectively) as well as the known
bifidogenic prebiotic fiber, OFS, allowed us to contrast the
effectiveness of these fractions on metabolic outcomes.
2. Methods and materials
2.1. Animals and diets
Male Sprague-Dawley rats (n = 100) were obtained from
Charles River (Montreal, QC, Canada) at aged 5 weeks and
housed in a temperature- and humidity-controlled room with
a 12-hour light/dark cycle. The rats were fed, ad libitum, a
high-fat/high-sucrose (HFHS) diet for 5 weeks to induce
obesity. The HFHS diet (19.2 kJ/g) consisted of (g/100 g): casein
(20.0), sucrose (49.9), soybean oil (10.0), lard (10.0), Alphacel
(5.0), AIN-93 M mineral mix (3.5), AIN-93 vitamin mix (1.0), DL-
methionine (0.3), and choline bitartrate (0.25) (Dyets Inc,
Bethlehem, PA, USA). After HFHS feeding, DIO rats were
selected as previously described [29], wherein the top 50th
percentile of weight gainers were randomized into the
intervention. Rats were fed ad libitum 1 of 5 isoenergetic
dietary treatments for 6 weeks (n = 10): (1) control (based on
AIN-93 M diet), (2) OFS (13% wt/wt to match fiber content in
diet, (3) yellow PF (30% wt/wt resulting in a 13% fiber diet),
(4) yellow PFL (30% wt/wt), or (5) yellow PS (30% wt/wt).
Although the PS product is more than 95% starch by weight,
the PF and PFL products are high in fiber (35% by weight) and
protein (23% by weight), respectively but contain starch,
protein, and fiber in varying concentrations, which was
taken into account in formulating the diets (complete
composition provided in Table 1). The PF, PFL, and PS diets
were formulated to derive 30% (by weight) of the overall diet
from each specific test ingredient. The OFS diet was used as a
 N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX 3

Table 1 – Ingredient composition of the diets fed to rats 2.4. Meal tolerance test
a
Composition (g/kg) Control OFS PF PFL PS
On the final day of the study (week 6 of treatment), a meal
Cornstarch 431.2 398.0 276.4 235.4 128.5
tolerance test (MTT) was performed according to our previous
Dextrinized cornstarch 143.5 146.9 137.0 139.5 144.3
work [29]. Rats were food deprived overnight. In the morning,
Casein 129.6 133.6 91.0 44.2 130.3
Sucrose 92.6 94.8 81.7 83.2 93.0 a baseline blood sample was collected followed by adminis-
Soybean oil 37.0 37.9 35.3 36.1 37.0 tration of a 3 g oral gavage of their respective diet emulsified in
Alphacel 120.4 28.4 35.1 117.1 121.0 3 mL water. Additional blood samples were collected at 15, 30,
AIN-93-MX 32.4 33.2 30.9 31.6 32.6 60, and 90 minutes postprandial with addition of Diprotin-A
AIN-93-VX 9.3 9.5 8.8 9.0 9.3 (0.034 mg/mL blood; MP Biomedicals, Irvine, CA, USA), Sigma
L-cystine 1.7 1.7 1.6 1.6 1.7
protease inhibitor (1 mg/mL blood; Sigma Aldrich, Oakville,
Choline bitartrate 2.3 2.4 2.2 2.3 2.3
Test ingredient b 0 113.6 300.0 300.0 300.0
ON, Canada), and Roche Pefabloc (1 mg/mL of blood; Roche,
Energy density (kJ/g) 13.8 14.2 13.8 13.8 13.8 Mississauga, ON, Canada). Plasma was stored at −80°C.
Carbohydrate (%kcal) 77.6 77.8 78.0 77.9 77.6
Fat (%kcal) 10.0 9.9 9.6 9.7 10.0 2.5. Tissue collection
Protein (%kcal) 12.4 12.4 12.4 12.4 12.4
a
Based on AIN-93 M purified diet with additional Alphacel to Immediately after the MTT, rats were overanaesthetized with
match energy density to experimental diets. isoflurane and killed via aortic cut. Cecal contents were
b
Pea starch as Accu-Gel Native Pea Starch (Nutri-Pea Limited, collected and stored at −80°C. The liver was excised weighed,
Portage la Prairie, MB, Canada); PFL as Fiesta Flour (Parrheim Foods, snap frozen, and stored at −80°C.
Saskatoon, SK, Canada); PF as Uptake 80 Functional Pea Fiber
(Nutri-Pea Limited); OFS as Orafti Raftilose P95 (Quadra Chemicals
2.6. Plasma analysis
Ltd, Burlington, ON, Canada). Pea starch composition (g/100 g): fat,
less than 0.1; starch, more than 95.0; protein, less than 1.0. Pea flour
composition (g/100 g): fat, 1.7; fiber, 7.1; starch, 54.2; sugars, 2.6; and Plasma leptin, insulin, ghrelin (active), and PYY (total) were
protein, 22.5. Pea fiber composition (g/100 g): fat, less than 0.5; fiber, measured in triplicate via multiplex (Rat Endocrine LincoPlex
more than 35.0; sugars, less than 2.5; starch, less than 50.0; and Kit; Millipore, St Charles, MO, USA) according to previous work
protein, less than 10.0. [31]. Active GLP-1 plasma concentrations were determined via
enzyme-linked immunosorbent assay (Millipore).

well-characterized prebiotic fiber comparison diet that mod-
ifies gut microbiota and improves metabolic outcomes
including gut satiety hormones [8,30]. The fiber content of
the OFS and PF diets were matched at a dose of 13%, and all
other diets were 12% to 13% fiber. Energy density and
macronutrient compositions were maintained across the 5
diets. Water was provided ad libitum and food intake
measured on 10 separate days throughout the 42-day
experiment by weighing feed cups to the nearest 0.1 g and
subtracting this weight from the previously measured weight.
The protocol was approved by the University of Calgary
Animal Care Committee and conformed to The Guide for the
Care and Use of Laboratory Animals.
2.2. Oral glucose tolerance test
One week before the end of the study (week 5 of treatment),
rats were food deprived for 12 hours. Baseline blood glucose
was measured via tail nick using a OneTouch glucose meter
(LifeScan Canada, Burnaby, BC, Canada). Rats were given an
oral glucose gavage (2 g/kg body weight) and blood glucose
measured at 15, 30, 60, 90, and 120 minutes postgavage.
2.7. RNA extraction and real-time polymerase chain reaction
Total RNA was extracted from the liver using TRIzol reagent
(Invitrogen, Carlsbad, CA, USA) as per manufacturers’ instruc-
tions. The concentration of RNA was determined using Ribo-
green (Invitrogen). Reverse transcription was performed with 1
μg of total RNA using the first strand complementary DNA
synthesis kit for real-time polymerase chain reaction (Invitro-
gen). Real-time polymerase chain reaction was performed
(BIO-RAD, Hercules, CA, USA) according to previous work [32]
with primers synthesized by the University of Calgary Core
DNA Services (Calgary, AB, Canada). Genes related to hepatic
fat metabolism including fatty acid synthase (FAS), acetyl coA
carboxylase (ACC), and sterol regulatory element-binding
protein 1c (SREBP-1c) were examined. Actin was verified as a
suitable reference gene and is not changed in response to the
treatments. Polymerase chain reaction amplification efficien-
cy was established for actin, FAS, ACC, and SREBP-1c by means
of calibration curves. Primers sequences have been previously
published [33]. Data were analyzed via the 2−ΔCT method [34].
2.8. Hepatic triglyceride content

2.3. Body composition
At the end of the study, rats were lightly anaesthetized with
isoflurane and body composition determined with dual
energy x-ray absorptiometry. Body weight, lean body mass,
fat mass, and bone mineral density were determined using
software for small animal analysis (QDR 4500; Hologic,
Bedford, MA, USA).
Triglycerides were measured using TG (GPO) reagent set (Point
Scientific, Canton, MI, USA) according to our previous work [35].
2.9. Gut microbiota profiling using quantitative
polymerase chain reaction
Microbial profiling was performed according to our previous
work [30]. Briefly, total bacterial DNA was extracted from cecal

Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
4 N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX

Table 2 – Food intake and body anthropometrics after 6 weeks of dietary treatment
Control PF PFL PS OFS
ab bc a abc
Average food intake (g/d) 25.1 ± 1.9 22.5 ± 2.8 29.8 ± 3.9 25.3 ± 2.1 18.8 ± 2.6c
Final body weight (g) 678.8 ± 17.4 664.8 ± 15.7 648.4 ± 11.9 675.3 ± 11.7 647.4 ± 16.6
Weight gain (g) 61.4 ± 3.0ac 61.3 ± 5.5a 44.2 ± 7.4b 72.5 ± 3.9c 51.1 ± 5.3ab
Lean mass (g) 503.4 ± 16.3 512.3 ± 10.1 507.9 ± 9.0 515.8 ± 13.1 505.1 ± 15.9
Fat mass (g) 175.4 ± 14.9a 152.5 ± 13.2ab 140.5 ± 13.7b 159.5 ± 8.4ab 143.9 ± 15.4b
Body fat (%) 26.9 ± 1.7a 24.0 ± 1.9ab 20.2 ± 1.4b 22.7 ± 0.96b 20.9 ± 1.7b
Liver weight (mg/g) 35.4 ± 2.9a 33.3 ± 1.6a 31.5 ± 1.9ab 30.9 ± 1.4ab 27.5 ± 3.7b

⁎Values are means ± SEM, n = 8 to 10. Average food intake represents the average of 10 separate days of food intake measured throughout the 6-
week experimental feeding period. Liver weight is presented as milligrams liver weight per gram body weight.
†Labeled means in a row without a common letter differ, P < .05. Oligofructose, PF, PFL, and PS.

samples using FastDNA Spin Kit for Feces (MP Biomedicals,
Lachine, QC, Canada) and quantified using PicoGreen DNA
quantification kit (Invitrogen). All samples were brought to a
concentration of 4 ng/μL before storage at −20°C for later
analysis. Amplification and detection were conducted in 96-
well plates with SYBR Green 2× qPCR Master Mix (BioRad).
Samples were run in duplicate with a final volume of 25 μL
containing 0.3 μmol/L primer and 20 ng template genomic
DNA. Group-specific primer sequences have been previously
published [16]. The 16S ribosomal RNA (rRNA) gene copies
value was calculated according the following webpage: http://
cels.uri.edu/gsc/cndna.html using average genome sizes.
Standard curves were normalized to the copy number of the
16S rRNA gene obtained from the following webpage: http://
rrndb.umms.med.umich.edu/ according to previous work
[16,36,37]. The percent Firmicutes was calculated as the sum
total of all Firmicutes (Clostridium coccoides, Clostridium leptum,
Clostridium cluster XI, Clostridium cluster I, Roseburia species,
and Lactobacillus species) divided by total bacteria.
2.10. Statistical analyses
All data are presented as a means ± SEM. Data were analyzed
via 1-way analysis of variance or where appropriate for
outcomes with multiple measures over time, repeated-
measures analysis of variance with Fisher least significant
difference post hoc analysis. Analysis of covariance (using
fasting levels of glucose or gut hormones [for oral glucose
tolerance test {OGTT}-related outcomes] and weight gain [for
gut microbiota and metabolic outcomes] as covariates] was
also performed. Total area under the curve (AUC) was
calculated for glucose concentrations during the OGTT and
for each of the hormones during the MTT. Incremental AUC
was also calculated for glucose. Differences were considered
significant at P ≤ .05. Statistical analyses were performed using
SPSS v16.0 software (SPSS Inc, Chicago, IL, USA).
PS, 550 ± 10.3; and OFS, 550 ± 10.3 g). Rats in the PF and
OFS groups had lower average food intake than the PFL group
(P = .05) (Table 2). Weight gain was significantly lower in PFL
compared with control, PF, and PS groups (P = .028). Rats fed PS
had the highest weight gain, which was significantly greater
than all other groups except control (P = .007) (Table 2).
Fat mass was lower in PFL and OFS compared with control
(P < .05). When expressed as percent body fat PFL, PS, and OFS
were all lower than control (P = .01). The OFS rats had lower
liver weight compared with control and PF (P = .046).
3.2. Oral glucose tolerance test
The PF group had decreased blood glucose concentrations at
time 0, 15, 60, and 120 minutes during the OGTT (P < .05)
and decreased total AUC compared with the control group (P =
.034) (Fig. 1A and B). When calculated as incremental AUC, PF
had significantly lower glucose incremental (iAUC) (355 ± 25.0
mmol/L × 90 minutes) compared with control (526.6 ± 47.9) and
PFL (455 ± 38.4 mmol/L × 90 minutes) (P = .032).
3.3. Meal tolerance test
There were no significant differences in insulin or GLP-1
during the 90-minute MTT (Fig. 1C-F). Oligofructose had
higher PYY at all time points during the MTT except
90 minutes and higher AUC compared with all other groups
(P < .001) (Fig. 1G and H). Pea flour (474 ± 43.1 pmol/L) had lower
fasting leptin concentration than control (751 ± 61.6 pmol/L)
and PF (750 ± 111 pmol/L) (P = .011). Leptin concentrations in
OFS (567 ± 43.1 pmol/L) and PS (677 ± 67.7 pmol/L)
were intermediate and not different from any other treat-
ments (P > .05). No significant differences were found in
ghrelin concentrations at individual time points or for AUC
(P > .05, data not shown).
3.4. Hepatic triglyceride content and gene expression

3. Results Pea flour had lower hepatic triglyceride content compared

3.1. Food intake and anthropometrics
After obesity induction, there were no significant differences
in body weight between the rats randomized to the 5
treatments (control, 552 ± 11.4; PF, 553 ± 13.5; PFL, 549 ± 11.0;
with PS but not the other groups (P = .036) (Fig. 2A). Acetyl coA
carboxylase messenger RNA (mRNA) levels were higher in OFS
vs PF, PFL, and control rats (P = .043) (Fig. 2B). Sterol regulatory
element-binding protein mRNA levels were higher in PF vs
OFS (Fig. 2C). No differences were detected in FAS mRNA levels
(data not shown).

Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
 N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX 5

Glucose AUC (mmol/L*90 min)

Plasma Glucose (mmol/L)
A 1200 B
13 * a ab ab ab
* b
11 * 900
9
* 600
7
5 Control OFS PF 300
PFL PS
3
0
0 15 30 60 90 120
Control OFS PF PFL PS
Time (min)

Insulin AUC (nmol/L*90 min)

Plasma Insulin (pmol/L)

600 C 45 D
500
36
400
27
300
200 18
100 9
0
0 15 30 60 90 0
Control OFS PF PFL PS
Time (min)
GLP-1 AUC (pmol/L*90 min)
Plasma GLP-1 (pmol/L)

12 E 1200 F
900
9

600
6
300

3 0
0 15 30 60 90 Control OFS PF PFL PS
Time (min)
PYY AUC (pmol/L*90 min)

45 †
Plasma PYY (pmol/L)

G 3000 H
37 a
† † 2400
29 †
1800 b
b b b
21 1200
13 600

5 0
0 15 30 60 90 Control OFS PF PFL PS
Time (min)

Fig. 1 – Concentrations of blood glucose (A) and total (tAUC) for blood glucose (B) during a 120-minute OGTT. Plasma insulin (C),
tAUC for plasma insulin (D), plasma GLP-1 (E), tAUC for plasma GLP-1 (F), plasma PYY (G), and tAUC for plasma PYY (H) during a
90-minute MTT in diet-induced rats consuming either control, yellow PF, yellow PS, OFS, or yellow PFL. ⁎PF different from
control (P < .05); †OFS different from all other groups (P < .05). Area under the curve values without a common superscript letter
differ, P < .05. Values are means ± SEM, n = 8 to 10.

3.5. Gut microbiota were increased with OFS compared with PF, PS, and control
(P < .05). C leptum (cluster IV) was significantly decreased with all
There was a significant bifidogenic effect of OFS but not the pea treatments (PF, PFL, PS, and OFS) compared with control (P < .05).
fractions compared with control (P < .001; Table 3). Lactobacilli C cluster I was increased with PS compared with all other groups

Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
6 N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX

A Triglycerides the PF group were likely driving a large part of the advantage
60 b they had in lower glycemia. After adjustment for fasting levels
Liver Triglycerides (mg/g) ab ab ab
a of insulin, GLP-1, and PYY, there remained a significant
difference in PYY AUC with OFS increasing PYY above all
40
other diets (P = .039). Similar to the nonadjusted analysis,
there remained no difference in insulin and GLP-1 AUC,
20 although there was a trend (P = .1) for GLP-1 AUC to be higher
in OFS compared with PFL and PS.
After adjustment for weight gain, percent body fat (P = .03)
0 but not fat mass (grams) (P = .062) remained different with PFL,
Control OFS PF PFL PS PS, and OFS having lower percent body fat than control. Liver
weight and liver triglyceride content were no longer signifi-
B ACC cantly different after adjustment for weight gain. The mRNA
30
mRNA Levels (Arbitrary

a ab levels of hepatic ACC and SREBP-1c, however, did remain

significantly different (P < .01) after controlling for weight gain.
20 b Adjusting for weight gain did not change the significant
b
Units)

b outcomes related to glucose and the gut hormones. Similarly,

controlling for weight gain did not alter any of the significance
10
associated with the gut microbiota except C cluster XI, which
was no longer significant at P = .09. All bacterial groups that
0 were significantly different before adjustment remained
Control OFS PF PFL PS significant after adjusting for weight gain including total
bacteria, Bacteroides/Prevotella species, C coccoides, C leptum,
C SREBP-1c C cluster I, Lactobacillus species, Bifidobacterium species,
30 a Methanobrevibacter species, percent Firmicutes, and the ratio
mRNALevels (Arbitrary

ab ab of Firmicutes:Bacteroidetes.

20
Units)

ab
b
10
0 and gut microbiota in DIO rats. The individual fractions
Fig. 2 – Liver triglyceride content (mg/g) (A), mRNA levels of
ACC (B), and mRNA levels of SREBP-1c (C) for diet-induced
rats consuming either control, yellow PF, yellow PS, OFS, or
yellow PFL. Values without a common superscript letter
differ, P < .05. Values are means ± SEM, n = 8 to 10.
(P < .05). C cluster XI was decreased with OFS compared with
control and PS (P < .05). Bacteroides/Prevotella species was
increased with OFS but not pea fractions compared with control
(P < .05). Methanobrevibacter was increased with PS compared
with all other groups (P < .05). The percent Firmicutes was
decreased with OFS, PF, and PFL compared with control (P = .05).
The ratio of Firmicutes:Bacteroidetes was reduced with OFS
compared with control and PS (P < .05) and reduced with PF and
PFL compared with PS (P < .05).
3.6. Adjusting for fasting glucose and hormone levels and
weight gain
After adjustment for fasting blood glucose, total AUC and
incremental AUC for glucose were no longer significant
suggesting that the lower fasting glucose concentrations in
4. Discussion
This study focused on evaluating the effects of yellow pea-
derived fractions (PF, starch, and flour) on metabolic outcomes
resulted in distinct effects on these outcomes. We accept the
hypothesis that PF improves glycemia and alters gut micro-
biota, most specifically a major member of the Firmicutes
phylum, C leptum. Beyond the PF fraction, however, other pea-
derived fractions also influenced the metabolic outcomes
examined. Weight gain varied upon exposure to the experi-
mental diets and was 28% lower in PFL rats than control. In
addition, the PFL group had the lowest percent body fat and
correspondingly lowest circulating leptin concentrations. Few
studies in rodents have addressed changes in body composi-
tion with pulses or their fractions, although dry beans have
been shown to reduce weight gain in DIO mice [38], and whole
and fractionated yellow PFL showed a trend to reduce body fat
in Golden Syrian hamsters [22]. In humans, consumption of
whole PFL reduced android:gynoid fat ratios in women
compared with white wheat flour, a finding not associated
with decreased food intake [25]. Although food intake in the
PFL rats was not different than control or PS, it was higher
than PF and OFS possibly implicating a change in energy
expenditure to explain the lower weight gain in PFL. This
would be consistent with findings from Marinangeli et al [22]
and Marinangeli and Jones [24] showing a change in the
thermic effect of food and altered substrate oxidation with
varying yellow pea fractions in humans. It is possible that the
protein content in the PFL may increase the thermogenic
effect of the diet and result in greater energy expenditure as
described by Marinangeli and Jones [23]. Arginine and

Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
 N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX 7

Table 3 – The 16S rRNA gene copies of cecal microbiota in obese rats after 6 weeks of dietary treatment
Control PF PFL PS OFS
b b ab b
Total bacteria 87 198 ± 1054 92 235 ± 1239 108 505 ± 8987 111 974 ± 1463 133 531 ± 1263a
Bacteroides/Prevotella 33 757 ± 482b 32 309 ± 350b 30 947 ± 413b 28 038 ± 463b 48 149 ± 605a
C coccoides 5469 ± 513 5403 ± 332 5761 ± 505 6449 ± 503 5478 ± 332
C leptum 4277 ± 716a 2553 ± 754b 2561 ± 325b 2310 ± 695bc 885 ± 169c
C cluster XI 13.2 ± 4.9a 6.6 ± 1.5ab 7.2 ± 3.0ab 13.3 ± 3.4a 3.3 ± 0.8b
C cluster I 43.1 ± 7.9b 49.9 ± 6.8b 49.9 ± 6.9b 89.4 ± 19.6a 32.1 ± 3.9b
Roseburia species 30.2 ± 11.6 5.5 ± 0.8 19.1 ± 9.5 16.3 ± 3.2 2.5 ± 0.9
Lactobacillus species 1367 ± 168bc 1158 ± 125c 1997 ± 350ab 1689 ± 215bc 2345 ± 241a
Bifidobacterium species 8.3 ± 0.5bc 9.6 ± 0.4bc 8.0 ± 1.0c 9.7 ± 0.5b 13.7 ± 0.3a
Methanobrevibacter species 0.5 ± 0.1a 0.4 ± 0.1a 0.4 ± 0.1a 0.7 ± 0.1b 0.4 ± 0.1a
Enterobacteriacea 1167 ± 171 1 112 129 1177 ± 215 1170 ± 150 1514 ± 325
Firmicutes (%) 62.9 ± 8.2a 44.4 ± 4.5bc 36.5 ± 5.4bc 50.6 ± 6.7ab 29.9 ± 3.4c
Bacteroidetes (%) 17.0 ± 2.2 15.4 ± 1.3 11.5 ± 1.6 11.8 ± 1.1 16.8 ± 3.0
Firmicutes:Bacteroidetes 4.3 ± 0.8ab 3.0 ± 0.4bc 2.9 ± 0.4bc 4.8 ± 0.8a 2.0 ± 0.2c

⁎Values are means ± SEM, n = 8 to 10 expressed as 16S rRNA gene copies/20 ng total genomic DNA. To fit in the table, all values, except the ratio
and percentages, were divided by 1000. Therefore, data are 16S rRNA gene copies (103)/20 ng genomic DNA.
†Because of unequal variance of the Bifidobacterium species values, the data were log transformed.
‡Labeled means in a row without a common letter differ, P < .05. Oligofructose, PF, PFL, and PS.

glutamine are 2 prominent amino acids in the protein
component of peas that have been shown to regulate energy
expenditure, potentially via increased mitochondrial biogen-
esis in the case of arginine [39,40]. Pulses also contain
nonnutrient bioactive substances or antinutrients (eg lectins
and enzyme inhibitors) that have been implicated in energy
regulation [19]. Although it might be tempting to suggest that
the presence of these substances in PFL reduced weight gain,
the level of these compounds is relatively low in yellow peas
compared with other pulses [41].
Although prebiotic fibers, such as inulin and OFS, have
consistently been shown to modulate gut hormone secretion,
our findings suggest that yellow pea-derived fractions may
influence metabolism and obesity via mechanisms distinct
from these appetite-regulating hormones. There were in fact
no changes in GLP-1, PYY, or ghrelin between the yellow pea
fractions. Similarly, Mollard et al [27] saw no change in GLP-1
or ghrelin with regular mixed pulse consumption (yellow
peas, chickpeas, navy beans, and lentils) in overweight and
obese humans. The lack of change in gut satiety hormones is
largely in keeping with our food intake findings, where only
the OFS group consumed significantly less energy than the
control group, and although PFL rats tended to eat more and
PF rats tended to eat less, this was not statistically different
from control.
Glycemia was examined at the end of the intervention, and
only yellow PF was able to significantly reduce glucose AUC
compared with control. This finding is consistent with a recent
study showing reduced fasting glucose in human subjects
consuming fractionated yellow PFL (ie, hulls only that are high
in fiber) [25] and most other studies examining the effect of
whole pulse consumption on glucose homeostasis [19]. It is
possible that some of the glucose-lowering effect seen in our
study may be due to the soluble fiber content of our specific PF,
which is relatively high (ie, 50% of the total fiber content).
Soluble fibers have been shown to improve postprandial
glycemic response by delaying gastric emptying and small
bowel transit, reducing enzymatic accessibility to substrate,
and slowing glucose absorption [42]. It is also noteworthy that,
after controlling for fasting blood glucose concentrations,
there was no longer a difference in the glucose AUC suggesting
that the lower baseline glycemia of the PF rats explained a
large portion of their glycemic advantage.
Inulin-type fructans, including OFS, alter the gut microbi-
ota and have a number of metabolic benefits including
enhanced satiety and improved glucose and insulin response
[43-45]. Some other dietary fibers such as those derived from
whole grain wheat have been investigated for similar bene-
ficial prebiotic effects to those of inulin-type fructans [46,47].
Based on the pea fractions tested in this study, we demon-
strated a reduction in the abundance of Firmicutes with PF,
PFL, and OFS compared with control. Although not unani-
mously shown, obesity is generally characterized by a an
increased Firmicutes:Bacteroidetes ratio [13,48]. Although the
relationship between obesity and gut microbiota is much
more complex than simply the ratio of Firmicutes:Bacteroi-
detes, a shift in these 2 major phyla has been linked to altered
energy extraction from carbohydrates in the colon.
Beyond the phylum-level changes, individual bacterial
groups were also differentially altered with the fractions.
Oligofructose is recognized for its ability to increase bifidobac-
teria whose beneficial role in the gut has been well established
[49]. Although PF and PS were able to increase bifidobacteria
abundance above that of control, this difference was not
significant and fell short of the bifidogenic effect of OFS. A
more prominent influence of the pea fractions on the gut
microbiota was in reducing C leptum, which was lower with all 3
fractions compared with control. C leptum is increased in obese
individuals [10]; however, their specific mechanism of action on
body weight is unclear. Recent work has shown that the
common bean, black beans, lentils, and chickpeas are an
excellent source of polysaccharides that can be fermented and
produce SCFA [19,50-52]. Short chain fatty acid are sensed by G
protein–coupled receptors in the intestine (GPR 41 and 43) [53]
and may be involved in regulation of appetite and adipose tissue
metabolism [54]. The changes in gut microbiota appear to occur

Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
8 N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX
independently of changes in weight gain given that statistically
controlling for weight gain did not alter the significant findings
related to gut microbiota outcomes.
There are some limitations to the current study. Although
we have identified some unique profiles of microbiota with the 3
pea fractions, future advanced metagenomic analysis would
provide insight into potential functional changes in the
bacterial in addition to the compositional shifts observed. In
addition, we did not use pair feeding in the experimental design,
which would have provided insight into which metabolic and
gut microbiota related outcomes were mediated independently
of changes in body weight. Statistically, we were able to perform
analysis with weight gain as a covariate to provide some insight
into the body weight independent effects.
Overall, this research demonstrates that diets enriched in
pulse-derived fractions appear to target different aspects of
metabolism including weight gain and fat mass, glucose
tolerance, and microbiota profiles. This work should form the
foundation for future examination of pulse-derived ingredients
for targeted aspects of metabolic dysfunction in humans.
REFERENCES
[1] WHO. Obesity and overweight fact sheet no. 311. WHO Media
Centre; 2013.
[2] Gross LS, Li L, Ford ES, Liu S. Increased consumption of
refined carbohydrates and the epidemic of type 2 diabetes in
the United States: an ecologic assessment. Am J Clin Nutr
2004;79:774–9.
[3] Anderson JW, Baird P, Davis Jr RH, Ferreri S, Knudston M,
Koraym A, et al. Health benefits of dietary fiber. Nutr Rev
2009;67:188–205.
[4] Deacon CF, Ahren B. Physiology of incretins in health and
disease. Rev Diabet Stud 2011;8:293–306.
[5] Karra E, Batterham RL. The role of gut hormones in the
regulation of body weight and energy homeostasis. Mol Cell
Endocrinol 2010;316:120–8.
[6] Cani PD, Neyrinck AM, Maton N, Delzenne NM. Oligofructose
promotes satiety in rats fed a high-fat diet: involvement of
glucagon-like peptide-1. Obes Res 2005;13:1000–7.
[7] Parnell JA, Reimer RA. Weight loss during oligofructose
supplementation is associated with decreased ghrelin and
increased peptide YY in overweight and obese adults. Am J
Clin Nutr 2009;89:1751–9.
[8] Delzenne NM, Cani PD, Daubioul C, Neyrinck AM. Impact of
inulin and oligofructose on gastrointestinal peptides. Br J
Nutr 2005;93:S157–61 [Suppl.].
[9] Schroeder N, Marquart LF, Gallaher DD. The role of viscosity
and fermentability of dietary fibers on satiety- and adiposity-
related hormones in rats. Nutrients 2013;5:2093–113.
[10] Angelakis E, Armougom F, Million M, Raoult D. The relationship
between gut microbiota and weight gain in humans. Future
Microbiol 2012;7:91–109.
[11] Schwiertz A, Taras D, Schafer K, Beijer S, Bos NA, Donus C,
et al. Microbiota and SCFA in lean and overweight healthy
subjects. Obesity 2010;18:190–5.
[12] Vrieze A, Van Nood E, Holleman F, Salojarvi J, Kootte RS,
Bartelsman JF, et al. Transfer of intestinal microbiota from
lean donors increases insulin sensitivity in individuals
with metabolic syndrome. Gastroenterology 2012;143:
913–7.
[13] Ley RE, Turnbaugh PJ, Klein S, Gordon JI. Human gut microbes
associated with obesity. Nature 2006;444:1022–3.
Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
[14] Cano PG, Santacruz A, Trejo FM, Sanz Y. Bifidobacterium
CECT 7765 improves metabolic and immunological
alterations associated with obesity in high-fat diet-fed
mice. Obesity 2013;21:2310–21.
[15] Yin YN, Yu QF, Fu N, Liu XW, Lu FG. Effects of four
bifidobacteria on obesity in high-fat diet induced rats. World J
Gastroenterol 2010;16:3394–401.
[16] Bomhof MR, Saha DC, Reid DT, Paul HA, Reimer RA. Combined
effects of oligofructose and Bifidobacterium animalis on gut
microbiota and glycemia in obese rats. Obesity 2014;22:
763–71.
[17] Parnell JA, Raman M, Rioux KP, Reimer RA. The potential
role of prebiotic fibre for treatment and management of
non-alcoholic fatty liver disease and associated obesity and
insulin resistance. Liver Int 2012;325:701–11.
[18] Wong JM. Gut microbiota and cardiometabolic outcomes:
influence of dietary patterns and their associated components.
Am J Clin Nutr 2014;100(Suppl 1):369S–77S.
[19] McCrory MA, Hamaker BR, Lovejoy JC, Eichelsdoerfer PE. Pulse
consumption, satiety, and weight management. Adv Nutr
2010;1:17–30.
[20] da S Queiroz-Monici K, Costa GE, da Silva N, Reis SM, de
Oliveira AC. Bifidogenic effect of dietary fiber and resistant
starch from leguminous on the intestinal microbiota of rats.
Nutrition 2005;21:602–8.
[21] Papanikolaou Y, Fulgoni VL. Bean consumption is associate
with greater nutrient intake, reduced systolic blood pressure,
lower body weight, and a smaller waist circumference in
adults: results from the National Health and Nutrition Survey
1999-2002. J Am Coll Nutr 2008;27:569–76.
[22] Marinangeli CP, Krause D, Harding SV, Rideout TC, Zhu F,
Jones PJ. Whole and fractionated yellow pea flours modulate
insulin, glucose, oxygen consumption, and the caecal
microbiome in Golden Syrian hamsters. Appl Physiol Nutr
Metab 2011;36:811–20.
[23] Marinangeli CP, Jones PJ. Pulse grain consumption and
obesity: effects on energy expenditure, substrate oxidation,
body composition, fat deposition and satiety. Br J Nutr 2012;
108(Suppl 1):S46–51.
[24] Marinangeli CP, Jones PJ. Chronic intake of fractionated
yellow pea flour reduces postprandial energy expenditure
and carbohydrate oxidation. J Med Food 2011;14:1654–62.
[25] Marinangeli CPF, Jones PJH. Whole and fractionated yellow
pea flours reduce fasting insulin and insulin resistance in
hypercholesterolaemic and overweight human subjects. Br J
Nutr 2011;105:110–7.
[26] Smith CE, Mollard RC, Luhovyy BL, Anderson GH. The effect of
yellow pea protein and fibre on short-term food intake,
subjective appetite and glycaemic response in healthy young
men. Br J Nutr 2012;108:S74–80 [Suppl].
[27] Mollard RC, Luhovyy BL, Panahi S, Nunez M, Hanley A,
Anderson GH. Regular consumption of pulses for 8 weeks
reduces metabolic syndrome risk factors in overweight and
obese adults. Br J Nutr 2012;108(Suppl 1):S111–22.
[28] Fernando WM, Hill JE, Zello GA, Tyler RT, Dahl WJ, Van Kessel
AG. Diets supplemented with chickpea or its main
oligosaccharide component raffinose modify faecal
microbial composition in healthy adults. Benef Microbes
2010;1:197–207.
[29] Eller LK, Reimer RA. Dairy protein attenuates weight gain in
obese rats better than whey or casein alone. Obesity 2010;18:
704–11.
[30] Parnell JA, Reimer RA. Prebiotic fibres dose-dependently
increase satiety hormones and alter Bacteroidetes and
firmicutes in lean and obese JCR:LA cp rats. Br J Nutr 2012;107:
601–13.
[31] Pyra KA, Saha DC, Reimer RA. Prebiotic fiber increases hepatic
acetyl CoA carboxylase phosphorylation and suppresses
glucose-dependent insulinotropic polypeptide secretion
 N UTR IT ION RE S EA RCH XX ( 2 0 14 ) XXX– X XX
more effectively when used with metformin in obese rats.
J Nutr 2012;142:213–20.
[32] Parnell JA, Reimer RA. Differential secretion of satiety
hormones with progression of obesity in JCR:LA corpulent
rats. Obesity 2008;16:736–42.
[33] Maurer AD, Chen Q, McPherson C, Reimer RA. Changes in
satiety hormones and expression of genes involved in
glucose and lipid metabolism in rats weaned onto diets high
in fiber or protein reflect susceptibility to increased fat mass
in adulthood. J Physiol Lond 2009;587:679–91.
[34] Silver N, Best S, Jiang J, Thein SL. Selection of housekeeping
genes for gene expression studies in human reticulocytes
using real-time PCR. BMC Mol Biol 2006;7:33.
[35] Parnell JA, Reimer RA. Effect of prebiotic fiber supplementation
on hepatic gene expression and serum lipids: a dose-response
study in JCR: LA-cp rats. Br J Nutr 2010;103:1577–84.
[36] Cowan TE, Palmnas MS, Yang J, Bomhof MR, Ardell KL, Reimer
RA, et al. Chronic coffee consumption in the diet-induced obese
rat: impact on gut microbiota and serum metabolomics. J Nutr
Biochem 2014;25:489–95.
[37] Reimer RA, Grover GJ, Koetzner L, Gahler RJ, Lyon MR, Wood S.
Combining sitagliptin/metformin with a functional fiber
delays diabetes progression in Zucker rats. J Endocrinol 2014;
220:361–73.
[38] Zhu Z, Jiang W, Thompson HJ. Edible dry bean consumption
(Phaseolus vulgaris L.) modulate cardiovascular risk factors
and diet-induced obesity in rats and mice. Br J Nutr 2012;108
(Suppl 1):S66–73.
[39] McKnight JR, Satterfield MC, Jobgen WS, Smith SB, Spencer
TE, Meninger CJ, et al. Beneficial effects of L-arginine on
reducing obesity: potential mechanisms and important
implications for human health. Amino Acids 2010;39:349–57.
[40] Iwashita S, Mikus C, Baier S, Flakoll PJ. Glutamine
supplementation increases postprandial energy expenditure
and fat oxidation in humans. JPEN J Parenter Enteral Nutr
2006;30:76–80.
[41] Campos-Vega R, Loarca-Pina G, Oomah BD. Minor compo-
nents of pulses and their potential impact on human health.
Food Res Int 2010;43:461–82.
[42] Silva FM, Kramer CK, de Almeida JC, Steemburgo T, Gross JL,
Azevedo MJ. Fiber intake and glycemic control in patients
with type 2 diabetes mellitus: a systematic review with
meta-analysis of randomized controlled trials. Nutr Rev 2013;
71:790–801.
[43] Blatchford P, Ansell J, de Godoy MRC, Fahey G, Garcia-Mazcorro
JF, Gibson GR, et al. Prebiotic mechanisms, functions and
applications. Int J Probiotics Prebiotics 2013;8:109–32.
Please cite this article as: Eslinger AJ, et al, Yellow pea fiber improves glycemia and reduces Clostridium leptum in diet-induced
obese rats, Nutr Res (2014), http://dx.doi.org/10.1016/j.nutres.2014.07.016
9
[44] Delzenne NM, Neyrinck AM, Cani PD. Gut microbiota and
metabolic disorders: how prebiotic can work? Br J Nutr 2013;
109:S81–5.
[45] Kellow NJ, Coughlan MT, Reid CM. Metabolic benefits of
dietary prebiotics in human subjects: a systematic review of
randomised controlled trials. Br J Nutr 2013:1–15 [Epub ahead
of print].
[46] Costabile A, Klinder A, Fava F, Napolitano A, Fogliano V,
Leonard C, et al. Whole-grain wheat breakfast cereal has a
prebiotic effect on the human gut microbiota: a double-blind,
placebo-controlled, crossover study. Br J Nutr 2008;99:
110–20.
[47] Napolitano A, Costabile A, Martin-Pelaez S, Vitaglione P,
Klinder A, Gibson GR, et al. Potential prebiotic activity of
oligosaccharides obtained by enzymatic conversion of durum
wheat insoluble dietary fibre into soluble dietary fibre. Nutr
Metab Cardiovasc Dis 2009;19:283–90.
[48] Ley RE, Backhed F, Turnbaugh PJ, Lozupone CA, Knight RD,
Gordon JI. Obesity alters gut microbial ecology. PNAS 2005;
102:11070–5.
[49] Russell DA, Ross RP, Fitzgerald GF, Stanton C. Metabolic
activities and probiotic potential of bifidobacteria. Int J Food
Microbiol 2011;149:88–105.
[50] Hernandez-Salazar M, Osorio-Diaz P, Loarca-Pina G, Reynoso-
Camacho R, Tovar J, Bello-Perez LA. In vitro fermentability
and antioxidant capacity of the indigestible fraction of
cooked black beans (Phaseolus vulgaris L.), lentils (Lens
culinaris L.) and chickpeas (Cicer arietinum L.). J Sci Food
Agric 2010;90:1417–22.
[51] Campos-Vega R, Garcia-Gasca T, Guevara-Gonzalez R, Ramos-
Gomez M, Oomah BD, Loarca-Pina G. Human gut
flora-fermented nondigestible fraction from cooked bean
(Phaseolus vulgaris L.) modifies protein expression associated
with apoptosis, cell cycle arrest, and proliferation in human
adenocarcinoma colon cancer cells. J Agric Food Chem 2012;60:
12443–50.
[52] Swiatecka D, Narbad A, Ridgway KP, Kostyra H. The study on
the impact of glycated pea proteins on human intestinal
bacteria. Int J Food Microbiol 2011;145:267–72.
[53] Brown AJ, Goldsworthy SM, Barnes AA, Eilert MM,
Tcheang L, Daniels D, et al. The orphan G protein-coupled
receptors GPR41 and GPR43 are activated by propionate
and other short chain carboxylic acids. J Biol Chem 2003;
278:11312–9.
[54] Soldavini J, Kaunitz JD. Pathobiology and potential
therapeutic value of intestinal short-chain fatty acids in gut
inflammation and obesity. Dig Dis Sci 2013;58:2756–66.