Journal of Cereal Science 64 (2015) 63e69

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Journal of Cereal Science

journal homepage: www.elsevier.com/locate/jcs

Evaluation of the functionality of ve different soybean proteins

in yeast-leavened pan breads

lez, Cristina Chuck-Hernandez, Sergio O. Serna-Saldvar*
Marco A. Lazo-Ve

gico de Monterrey-Campus Monterrey, Av. Eugenio Garza Sada 2501, Monterrey,
Centro de Biotecnologa- FEMSA, Escuela de Ingeniera y Ciencias, Tecnolo
N.L., C.P. 64849, Mexico

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 11 December 2014
Received in revised form
12 March 2015
Accepted 1 April 2015
Available online 9 May 2015

Soybean (SB) products are a source of proteins that complement the amino acid prole of cereal-based
products and consequently improve human health. Four different SB ours (SBF-1 to SBF-4) and a SB
concentrate (SBC) were incorporated into rened wheat our in order to increase approximately 20e25%
the protein content. The composite ours were processed into yeast-leavened pan breads. The SB fortied breads were characterized in terms of dough rheological, baking performance, bread crumb texture
and color and sensory properties. The different SB proteins affected differently rheological properties of
doughs, bread properties and quality. Addition of the SB proteins increased more than 3% optimum
dough water absorption and consequently bread yield but decreased between 7 and 13% bread volume.
The fortied breads also had a darker crumb. The best SB protein sources were SBF-3 and SBC which had
respectively 75.5 and 52% PDI, 24 and 36% NSI, 4.0 and 8.3 water absorption and 50.3 and 36.9% water
solubility indexes. These SB-fortied breads averaged 23% more protein and almost twice as much lysine
compared to the control. Therefore, these SB proteins can be utilized to produce yeast-leavened breads
with higher protein and upgraded protein quality.
2015 Elsevier Ltd. All rights reserved.

Keywords:
Bread
Soybean proteins
Functional properties
Dough rheological properties

1. Introduction
Soybeans (SB) are a rich source of high-quality protein and
nutraceuticals. The high protein content plus the high amounts of
essential amino acids that are lacking in most cereals make these
ours ideally suited to fortify cereal-based foods, with only a slight
increase in the production cost (Mashayekh et al., 2008; Novotni
et al., 2009). Furthermore, soybeans are rich sources of dietary ber, avonoids, isoavones, soyasaponins, other antioxidant compounds and most B-vitamins that exert positive health benets
especially in terms of prevention of most chronic diseases, osteoporosis and cancer (Mahmoodi et al., 2014).
Bread is the main staple in many countries worldwide and is
mainly prepared from rened wheat our. Nutritionally, the wide
array of white breads provide energy, proteins, minerals and
micronutrients (Shin et al., 2013; Acosta-Estrada et al., 2014), but
the nutritional quality of the protein is not adequate due to the low
levels of lysine present in wheat our. The partial replacement of
wheat our by protein rich ours is difcult because they do not

* Corresponding author. Tel.: 52 81 83284322; fax: 52 81 83284262.

E-mail address: sserna@itesm.mx (S.O. Serna-Saldvar).
http://dx.doi.org/10.1016/j.jcs.2015.04.007
0733-5210/ 2015 Elsevier Ltd. All rights reserved.

contain gluten-forming proteins and therefore are not functional,

especially in leavened-bread systems. In an early report, SernaSaldivar et al. (1988) produced SB enriched pan breads with and
without sodium stearoyl lactylate and concluded that this dough
conditioner improved volume and texture but not to the level of the
control bread. Likewise, Shin et al. (2013) manufactured SB-fortied
breads and concluded that they had comparatively denser texture
and the peculiar beany avor. However, SB proteins are potentially
suited to fortify bread, biscuits and other bakery formulations
especially in terms of enhancing protein quality and quantities of
relevant nutraceuticals (Serna-Saldivar et al., 1988; Mashayekh
et al., 2008; Ivanovski et al., 2012; Yezbick et al., 2013; Mahmoodi
et al., 2014).
Defatted SB our used in bread making increases absorption and
moisture retention, which enlarges the freshness or textural shelflife of the product. However, if a large amount of SB protein is
incorporated into the wheat our a disruption of the glutenforming proteins occurs. Therefore, the SB protein interferes with
the starchegluten matrix negatively affecting volume, crumb
scores and overall quality attributes and acceptability of enriched
breads (Shin et al., 2013; Mahmoodi et al., 2014).
The aim of this research was to assess the effect of addition
of four different defatted SB ours or a SB concentrate with

64

M.A. Lazo-V
elez et al. / Journal of Cereal Science 64 (2015) 63e69

contrasting functional properties on the rheological dough properties and quality of yeast-leavened pan breads produced by the
pup loaf straight or sponge dough processes. Control and SB fortied treatments were compared in terms of chemical composition,
amino acids, dough rheological properties, bread texture and sensory attributes.
2. Materials and methods
2.1. Commercial soybean samples
Five different commercial SB samples were selected to differ in
functional properties especially in terms of urease activity, water
absorption (WAI), water solubility (WSI), protein dispensability
(PDI) and nitrogen solubility (NSI) indexes. Four were defatted SB
ours, Industrial de Alimentos (SBF-1), GAF-120 (SBF-2), ADM (SBF3) and Ragasa (SBF-4) containing approximately 49% protein (N x
6.25 as is basis), whereas the remaining a protein-concentrate (SBC)
Provita containing 67% protein (as is basis). Urease activity
(Method Ba 9-58, AOCS, 2011), nitrogen solubility (Method Ba 1165, AOCS, 2011), water absorption and solubility (Cheftel et al.,
1989), and protein dispersibility (Method Ba 10b-09, AOCS, 2011)
indexes were assayed. Lysine, tryptophan and sulfur containing
amino acids (methionine plus cysteine) in wheat our and the
different SB protein sources were determined according to Ofcial
Method 982.30 E (a,b,c) of the AOAC (2006).
2.2. Soybean-fortied ours
Each of the ve different SBF proteins were composited with
commercial rened wheat our (La Perla, Molinos del Fenix, Saltillo, Coahuila, Mexico) in order to increase the protein concentration to about 20e25%. The experimental SB enriched ours were
produced by substitution of 6.0% SBF-1; 6.1% SBF-2; 6.3% SBF-3,
6.2% SBF-4 and 4.5% SBC of the rened bread our. Protein
(N  5.7) was determined in the control and experimental composite our samples using the AACC International (2000) method
46-30.
2.3. Dough rheological properties
The dough rheological properties of the control and SB-composite
ours were determined with the farinograph (Brabender Instruments,
South Hackensack, NJ) and Alveograph (Chopin Instruments,
Villeneuve-La-Garenne, France) according to Approved Methods 5421 and 54-30, respectively (AACC International (2000)).
2.4. Straight dough baking
The pup loaf straight-dough bread micro-baking method 1010.03 (AACC International, 2000) was utilized. The bakers formulation consisted of 6% rened cane sugar (Avance, Avance Comercial
de Monterrey, Monterrey, NL, Mexico), 3% vegetable shortening
(Inca, Unilever de Mexico S.A de C.V., Tultitl

an, Edo. de Mexico,
Mexico), 2% rened iodinated salt (La Fina, Sales del Istmo, Coatzacoalcos, Veracruz, Mexico) and 2% dry yeast (Saccharomyces cerevisiae) (Azteca Levadura, Iztapalapa, Mexico, D.F., Mexico).
Optimum water absorption and mix times were subjectively
determined by observing dough properties or gluten development
(lm formation, gloss and dough stickiness). Bake absorption,
mixing time, proof height, loaf height, oven spring, loaf weight, loaf
volume, and loaf apparent density were determined. Proof height
and loaf height were determined with a proof height meter
(National Manufacturing Co., Lincoln, Nebraska). The difference
between these values was recorded as oven spring. Loaf volume was

determined by rapeseed displacement (National Manufacturing Co.,

Lincoln, Nebraska) according to method 10-05.01 of the AACC
International (2000).
2.5. Sponge dough baking
Sponge dough breads were manufactured in a pilot plant in
order to generate loaves for texture and sensory analyses. Sponge
doughs were produced by mixing by hand 200 g of rened bread
our, 5 g instant dry yeast and 140 g water. The resulting blend was
placed in a plastic container in a fermentation cabinet (National
Mfg., Lincoln, NB, USA) set at 28 1  C for 4 h. Resulting sponges
were mixed with the remainder of the dry ingredients: 300 g bread
our, 30 g sugar, 20 g shortening, 5 g of dry whole milk (Nestle de
Mexico, Mexico, D.F., Mexico), 10 g salt, 1 g lecithin (Proveedores de
Ingeniera Alimentaria S.A. de C.V. Monterrey, N.L., Mexico), 1 g
Sodium Stearoyl Lactylate (SSL), 1 g calcium propionate and 2.5 g
dry yeast instant for one min at low speed in a Hobart mixer
equipped with the hook attachment. Then, the rest of the predetermined amount of water was added and blended for one
minute at low speed. Next, the velocity was switched to medium
until attaining optimum dough development. Film formation, gloss
and dough stickiness were subjectively determined to estimate
optimum mix times. Resulting doughs were weighed before placing
them in a fermentation cabinet set at 28 1  C and 85% relative
humidity. After 10 min resting, doughs were punched thru 0.95 cm
roll spacing in preparation for molding and panning in 7.5 cm
height metal pans that had the following dimensions on the base
and top, respectively: 22.5 and 24.5 cm long and 8.5 and 10.5 cm
wide. Baking pans were previously greased with vegetable shortening on the bottom and sides. Panned doughs were proofed for
45 min before baking for 25 min in an oven (Electrolux EOG Gas
single oven X 601) set at 190  C. Upon 30 min cooling at room
temperature, breads were cut into 2 cm thick slices, packaged in
sealed polyethylene bags and stored at room temperature for 5
days.
2.6. Crumb texture and color of sponge breads
The crumb texture of the slices of bread (2 cm thick) was evaluated with a texture analyzer (model TA.XT plus, Stable Micro
systems, United Kingdom) according to the bread rmnesscompression test method 74-10.02 (AACC International (2000))
with a trigger force of 0.048 N. Cohesiveness, hardness, gumminess,
chewiness and elasticity were evaluated at the center of the slice.
The parameters were calculated from the resulting texture prole
analysis curves. Tests were conducted at days 0 (fresh), 1, 2 and 5
days of bread slices kept at room temperature.
The crumb color of the control and experimental breads was
objectively measured on three slices with a colorimeter (CR 300,
Minolta, Japan). Bread-crumb color parameters L*, a*, and b* were
the average of three measurements at different parts of the slice.
Tests were conducted after 1 day storage at room temperature.
2.7. Sensory analyses
In-house consumer panels (pilot consumer panels) consisted of
forty untrained panelists, evaluated the sensory features and
overall acceptability of control and experimental breads in individual booths after 24 h of baking. Bread evaluation was performed
in a sensory evaluation laboratory (ITESM-Campus Monterrey)
according to the guidelines described by Watts et al. (1989). Each
panelist was simultaneously given six coded samples along with a
ballot, and was asked to rate color, texture, avor, odor, and overall
quality on a 5-point hedonic scale, where 1 is like very much and

M.A. Lazo-V
elez et al. / Journal of Cereal Science 64 (2015) 63e69

5 is dislike very much. In order to obtain an overall evaluation of

the samples, a score was assigned to each bread for nutritional,
dough properties, baking performance and sensory acceptability
parameters.
2.8. Overall evaluation of breads
In order to obtain an overall evaluation of the samples, a score
was assigned to each bread for nutritional, dough properties,
baking performance and sensory acceptability parameters. The
control bread scored 1 for each property and considered these
values as baseline. The experimental SB-fortied breads were
assigned with neutral, positive (better) or negative (worse) values
according to variations among means (Tukey tests at p < 0.05). For
instance, when the parameter mean was higher or lower than one
or two standard deviations compared to the corresponding mean of
the control, the individual scores assigned were 1 and 2 or 1
and 2, respectively. Final desirability scores were the addition of
all individual scores.
2.9. Statistical analysis
Each experiment was performed in triplicate in three different
weeks and data was reported as means standard deviations.
Results were subjected to analysis of variance (ANOVA) and differences among means were compared by Tukey tests at p < 0.05.
All computations were made by the statistical software JMP
(version 11).
3. Results and discussion
3.1. Protein and amino acid composition
As planned, the protein content of the different SB-composite
ours were higher compared to the control and similar among all
the experimental ours. These averaged 14.4% protein (dry weight
basis), representing an average increase of 23% compared to the
control wheat our (11.7% protein dry weight basis).
More importantly, these SB products contained between 11 and
16 times higher lysine contents (3.45e4.93%) compared to the
rened wheat our (0.31%) (Table 1). Therefore, the addition of
approximately 6 and 4.5% of the different SB defatted ours and
concentrate respectively increased 1.6 times the lysine content in
composite ours and breads (Table 1). The contrasting difference
between the rened wheat our and the SB protein sources especially in terms of lysine makes these proteins ideally suited to upgrade protein quality and nitrogen retention. Earlier studies clearly

65

documented that the higher protein and enhanced essential amino

acid scores of SB-fortied breads improved protein quality and
animal growth (Serna-Saldivar et al., 1988; Mashayekh et al., 2008;
Mahmoodi et al., 2014). The limiting essential amino acid in wheat
is lysine, which is present in about half the amount required for
optimum infant growth. Nutrition studies have clearly indicated
that the higher lysine improves the amino acid score, protein efciency ratio, nitrogen retention evaluated by the biological and net
protein utilization values, growth, brain development and memory
performance in laboratory animals and humans (Serna-Saldivar
et al., 1988; Amaya-Guerra et al., 2006; Khan et al., 2009; Serrem
et al., 2011; Mahmoodi et al., 2014).
The SBF-1 showed the lowest protein dispersibility (PDI) and
water solubility (WSI) indexes. On the other hand, SBF-3 and 4 had
about 15 times higher urease activity and 3 times higher PDI
compared to SBF-1 or 2. SBF-3 and 4 had high urease activity and
PDI values due to the slight heat treatments applied prior to oil
extraction and during meal desolventization in a desolventizertoaster or DT. The PDI reported for SBC is similar to the value reported by Wang et al. (2004) for a counterpart produced from acidwashed white akes.
3.2. Rheological properties of SB-enriched doughs
3.2.1. Farinograms
As anticipated, rheological properties of doughs estimated with
the farinograph and baking performance were affected by the
addition of the different SB protein sources (Tables 2 and 3).
Addition of the different SB sources improved the dough water
absorption capacity by 3.5 units which represented about 6% more
water absorption compared to the wheat our (Table 2). Compared
to the wheat our, the corrected water absorption was between 2.2
and 4.7% units higher for the different SB-enriched counterparts.
Several investigations had documented the effect of increased
water absorption in doughs supplemented with different SB ours
(Serna-Saldivar et al., 1988; Ribotta et al., 2005). These differences
are related to the nature of the soybean proteins which are hydrophilic or with high WAI (Table 1). WAI of the different SB proteins ranged from 4.0 in the high PDI SBF-3 to 5.3 in the lowest PDI
SBF-1.
Addition of all SB ours prolonged arrival times, the longest
times were observed in composite ours containing SBF-2 or SBF-4.
Interestingly, these meals had low NSI values (Table 1). Only the
composite our containing 4.5% SBC had similar arrival time
compared to the control our (Table 2). The arrival times were
inversely correlated with both nitrogen solubility (NSI) (r 0.57)
and water absorption indexes (WAI) (r 0.56) (Table 4). In

Table 1
Functional characteristics and main essential amino acids of ve different soybean protein sources.a
Parameter

Crude Protein % (N  6.25, dry basis)c

Amino acid (%, dry weight basis)d
Lysine
Tryptophan
Methionine Cysteine
Urease activity
Nitrogen Solubility (NSI, %)
Water Absorption Index (WAI)
Water Solubility Index (WSI)
Protein Dispersibility Index (PDI)
a
b
c
d

Soybean protein sourcesb

SBF-1

SBF-2

SBF-3

SBF-4

SBC

53.6 0.2C

54.582B

54.58 0.4B

53.6 0.1C

74.9 0.2A

3.45
0.85
1.56
0.10 0.01E
19.7 0.7B
5.3 0.0B
25.7 0.4E
23.2 0.1D

3.5
0.87
1.61
0.15 0.01D
13.4 0.8C
4.8 0.3C
28.4 0.1D
25.3 1.6D

3.54
0.82
1.64
2.25
24.0
4.02
50.3
75.5

3.55
0.76
1.63
2.20 0.03B
25.5 0.7B
4.3 0.1D
47.6 1.8B
66.9 1.5B

4.93
1.09
1.95
0.40 0.01C
36.1 1.5A
8.3 0.1A
36.9 0.2C
52.0 0.5C

0.01A
0.2B
0.0E
0.2A
1.7A

Averages standard deviations, values with the same letter within row are not signicantly different at p < 0.05, (n 3).
The experimental SB enriched ours were produced by substitution of 6.0% SBF-1; 6.1% SBF-2; 6.3% SBF-3, 6.2% SBF-4 and 4.5% SBC of the rened bread our.
The rened hard wheat our contained 11.72% crude protein (N  5.7, dry basis).
The wheat our contained 0.31, 0.18 and 0.50% (dry weight basis) of lysine, tryptophan and methionine cysteine, respectively.

M.A. Lazo-V
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66

Table 2
Effect of soybean protein addition on the rheological properties of doughs estimated with the farinograph and alveograph.a
Parameter

Flour enriched withb

Control
Wheat our

SBF-2

12.2 0.2

10.1 0.4

Flour fortied protein, % (14% mb)

Farinograph
Water Absorption (%)
Corrected Water Absorption (%)
Arrival Time (min)
Departure Time (min)
Dough Stability (min)
Dough Development (min)
Mixing Tolerance Index (BU)c
Alveograph
P (mm H2O)
L (mm)
P/L
G
W (104J)
b

SBF-1

SBF-3

12.3 0.7

SBF-4

12.4 1

SBC

12.3 0.7

12.5 0.1A

58.6
58.3
0.9
13.5
12.7
8.0
40

0.2C
0.2C
0.1B
0.7A
0.6A
0.2A
0B

63.4
63.04
1.3
11.5
10.2
7.0
50

0.3A
0.3A
0.1B
0.4B
0.4B
0.3C
0AB

61.6
60.49
3.9
9.3
5.4
7.0
55

0.2B
0.2B
0.5A
0.4C
0.1C
0.4C
7.1A

61.0
60.5
1.25
14.1
12.9
8.0
30

0.1B
0.1B
0.1B
0.1A
0.1A
0.2A
0C

63.4
62.98
3.8
11.4
7.7
7.5
40

0.1A
0.1A
1.1A
0.1B
1.2C
0.2B
0B

61.8
61.5
0.8
11
10.3
6.5
50

0.2B
0.2B
0B
0B
0B
0.4D
0AB

123.5
53.5
2.3
16.3
275

4.9B
2.1A
0.2C
0.2A
2.8A

172
36
4.9
13
270

7.1A
0.7B
0.3ABC
0.1B
7.8A

120.5
19
6.6
9.7
101

4.9B
5.7C
1.7A
1.3C
20.5B

138.5
48
2.9
15.0
278

4.9B
3.5A
0.3BC
0.6AB
7.1A

134.5
52
2.6
16
288

0.7B
0A
0C
0.1A
2.1A

183.5
35.5
5.2
13.3
293

2.1A
0.7B
0.1AB
0B
5.7A

Average standard deviations, values with the same letter within row are not signicantly different at p < 0.05, (n 2).
The experimental SB enriched ours were produced by substitution of 6.0% SBF-1; 6.1% SBF-2; 6.3% SBF-3, 6.2% SBF-4 and 4.5% SBC of the rened bread our.
Brabender or Farinograph Units.

Table 3
Effect of soybean protein addition on the dough mixing and baking properties of pup loaves produced with the straight dough micro-baking procedure.a
Treatment

Flour enriched withb

Control
Wheat our

Water Absorption (%)

Dough Mixing Time (min)
Oven Springc (cm)
Bread Weight (g)
Bread Volume (cm3)
App. Bread Densityd (g/cm3)
a
b
c
d

63.7
3.9
2.3
145
899.2
0.16

0.5D
0.13BC
0.38A
0.7BC
20A
0.1C

SBF-1
65.2
4.1
1.5
146.7
783.5
0.19

SBF-2

0.3C
0.1AB
0.2B
1.8ABC
34C
0.1AB

66
4.0
1.5
149.4
775.8
0.19

SBF-3

0.8B
0.0AB
0.39B
1.7AB
33C
0.1A

64.7
4.4
1.5
144.5
809
0.18

SBF-4

0.5C
0.34A
0.44B
2.6C
27BC
0.0B

65.3
4.4
1.6
151.1
818.3
0.18

SBC

0.3BC
0.46A
0.61B
5.3A
9.3BC
0.0AB

67.3
4.1
1.7
146.6
828.3
0.18

0.3A
0.34AB
0.28B
2.1ABC
20B
0.0B

Averages standard deviations, values with the same letter within row are not signicantly different at p < 0.05, (n 6). Temperature of fermentation 31.2 0.8  C.
The experimental SB enriched ours were produced by substitution of 6.0% SBF-1; 6.1% SBF-2; 6.3% SBF-3, 6.2% SBF-4 and 4.5% SBC of the rened bread our.
Oven spring bread height e proof height.
Apparent Bread Density.

contrast, the departure time was positively correlated with both

WSI (r 0.73) and PDI (r 0.72). These data indicated that the
quantity of soluble proteins and PDI increased departure time.
According to Maforimbo et al. (2008), the poor dough and baking
qualities of composite wheat-soybean formulations could be
explained by the fact that wheat proteins favor hydrophobic

Table 4
Correlation between soybean quality parameters and rheological properties, and
micro-baking test estimated with different SBFs.
Parameters
Farinograph
Water Absorption (%)
Arrival Time (min)
Departure Time (min)
Dough Stability (min)
Dough Development (min)
Mixing Tolerance Index (BU)2
Alveograph
P (mm H2O)
L (mm)
P/L
G
W (104J)
Baking properties
Water Absorption (%)
Dough Mixing Time (min)
Oven Spring (cm)
Bread Volume (cm3)

%NS

WAI

WSI

PDI

0.05
0.57
0.30
0.49
0.26
0.22

0.12
0.55
0.28
0.13
0.83
0.48

0.33
0.06
0.73
0.49
0.73
0.91

0.27
0.12
0.72
0.51
0.63
0.88

0.67
0.47
0.33
0.52
0.76

0.79
0.29
0.45
0.22
0.22

0.26
0.79
0.84
0.77
0.49

0.14
0.82
0.85
0.81
0.59

0.53
0.26
0.89
0.93

0.90
0.52
0.76
0.41

0.34
0.91
0.16
0.67

0.24
0.89
0.30
0.77

The bolded data had signicant Pearson correlations (p < 0.05).

interactions whereas SB proteins had more afnity for water or are

hydrophilic.
Dough development times were similar (p > 0.05) for the control and SBF-3 composite dough and lowest for the SBC dough
(Table 2). The dough development times were positively correlated
with WSI (r 0.73) and PDI (r 0.63), and showed a high negative
correlation with WAI (r 0.83). The soluble proteins related to
PDI affected the time to achieve maximum consistency likely due to
the gel-forming ability of these macromolecules.
Dough stability times in all SB-enriched ours, except for the
one containing SBF-3, were lower compared to the control. The
farinograph dough stabilities were around 20% less for composite
ours with SBF-1 and SBC, 40% less for the SBF-4, and 57% less for
SBF-2. Ribotta et al. (2005) observed that both dough development
and stability times decreased in doughs prepared with mixtures of
wheat our with SB (active or heat-treated). In conclusion, the
highest PDI (Table 1) SBF-3 was the protein source that impacted
the most of all farinograph parameters. The two best performing SB
sources (SBC and SBF-3) had farinograph dough development and
stability times of 6.5e8 and 10.3e12.9 min, respectively. The correlation between farinograph dough stability time and PDI
(r 0.51) was moderate whereas correlations between mixing
tolerance index and PDI (r 0.91) and WSI (r 0.88) were
negative. The mixing tolerance indexes in all experimental doughs
showed varying tolerances in relation to the control. The our
containing SBF-2 with a low PDI yielded the weakest dough. This
particular dough had 37.5% reduced strength followed by counterparts containing SBF-1 or SBC which were approximately 25%

M.A. Lazo-V
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weaker. Interestingly, the composite our with SBF-4 had similar

strength compared to the control whereas the our with SBF-3
increased its strength by 25%. These values analyzed jointly with
the dough stability indicated that the our enriched with SBF-2 was
unstable and less suitable for baking (MTI > 50 BU and dough
stability time of only 5.4 min). On the other hand, SBF-3 had the
best farinograph dough rheological properties (MTI < 30 BU and
dough stability time of 12.9 min) for bread making.
3.2.2. Alveograms
Alveograms also demonstrated that the different SB proteins
affected differently the properties of doughs. In terms of alveograms (Table 2), composite ours containing SBC or SBF-1 had a
higher maximum alveograph over pressure compared to the wheat
our whereas the composite our containing SBF-2 showed the
lowest P value. The highest extensibility dough values were
observed in the control and high-PDI SBF-3 and 4 composite ours.
This indicates that the low heat-treated ours favored extensibility
to a level similar to the control wheat our. These notorious differences in P and L values affected the P/L ratios that are known to
be closely related to our functionality. The composite ours that
had similar values to wheat our were the composite ours containing SBF-3 and 4. As expected and due to the addition of glutenfree SB proteins, the index of swelling (G), that gives an indication
of the work to extend the dough, was signicantly lower for all
composite ours except for the system containing SBF-4. The best
performing composite ours produced with SBC or SBF-3 had G
values between 13.3 and 15 and W values between 293 and 278
*104 J (Table 2). Therefore, these doughs were suitable for yeastleavened bakery purposes (W values ranging from 200 to 300
*104 J). These G and W values were similar to the control dough.
Addition of the SBF-2 to wheat our negatively inuenced most of
the alveograph parameters. This particular composite our had the
lowest G and W values. In fact, this our with a W value of only
101  104 J was in the range suited for pastries (60e120 W).
The alveograph W values showed a moderate positive correlation with NSI (r 0.52) and PDI (r 0.60). The PDI has been proposed as a quality parameter to appraise heat treatment processes
applied to legume ours during their production in the oil crushing
industries. Soybean ours having PDI values higher than 50 were
the most recommended for bread making (Table 1).
3.3. Baking properties of soybean enriched pup loaves
The SBF-3 and SBC with adequate alveograph W values and
farinograph parameters produced the best straight dough breads in
terms of bread volume, height and apparent density (Table 3). The
optimum dough water absorptions assayed in the straight microbaking procedure of all composite ours were signicantly higher
compared to the control (p < 0.001). As similar as observed in
farinograms (Table 2), the SB-enriched doughs absorbed on average
3.2% more water compared to the control (Table 3). Among the
different SB composite ours, the SBC absorbed the highest (67.3%)
water, despite it being the source added with the least amount
(only 4.5%). Similar results were previously reported by SernaSaldivar et al. (1988). Soybean proteins are rich in hydrophilic and
binding protein moieties which increase water absorption. As a
result, the SB fortied yeast-leavened breads usually contain more
moisture and retain longer their crumb softness (Serna-Saldivar
et al., 1988; Sathe, 2012).
On the other hand, a high positive correlation was observed
between dough water absorption and WAI and a moderate positive
correlation with NSI (Table 4). Thus, the composite our containing
SBC with the highest WAI (8.34) showed the highest our water

67

absorption. Interestingly, the SBC had approximately 50% more WAI

compared to the SBF defatted ours tested herein (Table 1).
Optimum dough mixing times estimated during the microbaking tests were also affected by the addition of the different SB
proteins with values uctuating from 4.0 to 4.4 min. Doughs produced with SBF-3 and 4 (higher PDI) required longer dough mixing
schedules (4.4 min) compared to the control (3.9 min). Doughs with
SBF-1, SBF-2, or SBC had intermediate dough mixing requirements.
Dough mixing times were highly correlated with WSI (r 0.91) and
PDI (r 0.90). It is well known that high PDI and WSI soybean
ours contain high amounts of water soluble-proteins and resulting doughs commonly require higher mixing times due to their
lower dough integration and the thickening effect. On the other
hand, doughs manufactured with SB with low NSI (<20%) values
had evident problems with integration and yielded sticky doughs.
This was especially observed with the composite our containing
SBF-2 (Table 1). Interestingly, the SBF-3 or SBC did not present
major integration and textural problems during the dough mixing
stage. These two SB proteins had high PDI (>52) and NSI (>24%)
values (Table 1). Thus, a correct balance between NSI and PDI are
crucial for the selection of the most appropriate SB proteins in order
to generate appealing and good quality yeast-leavened breads. Low
NSI values indicate that the protein is denatured to a higher extent
which leads to water insolubilization and the formation of protein
aggregates (Ribotta et al., 2005). Thus, longer dough mixing times
might be due to inefcient dough integration, especially during the
rst minutes of kneading. Moreover, a moderate inverse correlation
(r 0.52) between WAI and optimum dough mixing time was
attributed to the high hydration capacity of the SBF proteins. It is
well-known that SB has high concentrations of hydrosoluble proteins (65e75%) compared to cereals (25%) (Serna-Saldivar, 2010).
Both oven spring and bread volume were negatively affected by
the different SB sources (p < 0.0001) (Table 3). Flours containing
SBC, SBF-4 or SBF-3 yielded breads with volumes higher than
800 cm3; however, only the our containing SBC exceeded the
820 cm3 considered as optimal. The gluten strength and dilution
explain the lower oven-spring values observed in all experimental
breads The oven spring had a positive correlation with WAI
(r 0.76) and with NSI (0.89) (Table 4). Dough viscosity has a
water-release effect necessary for starch gelatinization during
baking (Shin et al., 2013) and expedited yeast proliferation with a
consequent better gas generation and retention that positively
affected bread volume. Therefore, higher our water absorptions
generally yield better bread volumes. However, the slightly lower
bread volume observed in SB enriched breads is mainly attributed
to the addition of a non-gluten forming protein and the loss of
gluten interaction. It is also known that incorporation of single-cell
protein can disrupt the elastic gluten structure, allowing losses of
gas during proong and baking (Fleming and Sosulski, 1978).
Ribotta et al. (2005) showed that different SB ours and proteins
produced a gluten lm which was more permeable to the CO2
generated by yeast. Fleming and Sosulski (1978) showed that the
loss of gas during baking may be through small pores in the
viscoelastic gluten lm protein observed by scanning electronmicroscopy in breads supplemented with SB protein concentrate.
Soybean enriched doughs produced from denatured and partially
denatured proteins yielded weaker gluten lms compared to
counterparts produced from less denatured sources. These observations are in accordance to Ribotta et al. (2005).
3.4. Sensory analyses and overall evaluation of breads
Sensory evaluation results revealed that panelists did not
perceive signicant differences in color, odor, avor, texture and
overall acceptance of all breads with scores ranging from 1.5 to 1.8

68

M.A. Lazo-V
elez et al. / Journal of Cereal Science 64 (2015) 63e69

Fig. 1. A. Effect of soybean protein addition on the sensory properties of pan breads using a 5-point hedonic scale (1 like very much and 5 dislike very much) and B. Texture
parameters of slices of fortied breads throughout ve days storage at room temperature. Values with the same letter within cluster are not signicantly different at p < 0.05.

(p 0.09), 1.8-2.2 (p 0.54), 1.8-2.1 (p 0.32), 1.8-2.1 (p 0.64) and

1.7-2.1 (p 0.26) (Fig. 1A), respectively. Although rated equally,
panelists documented changes in crumb colorations in the SBenriched breads. These breads had a darker cream coloration
compared to the light cream color of the control or SBC enriched
crumbs. As expected, among the different SB enriched breads the
one manufactured with SBC had the lightest crumb color. All SB
enriched breads also had darker crust colors compared to the
control. These color changes have been previously documented in
other investigations (Mashayekh et al., 2008; Ivanovski et al., 2012;
Yezbick et al., 2013). Thermal non-enzymatic caramelization and
Maillard reactions occurring between reducing sugars and amino
acids are the main ones responsible for the observed darker crust
colorations (Ivanovski et al., 2012). Thus, the observed changes in

the crust are related to the non-protein fraction of the different SBF
proteins associated with the NSI.
Objective crumb bread lightness L* and a* values were not
signicantly different among breads, whereas b* values were
similar among breads manufactured with SB having high and low
PDI. Also, bread crust lightness (L*) values were different in all
breads whereas a* values were similar (Supplementary Fig. 1). On
the other hand, bread cohesiveness, hardness and chewiness,
evaluated by the texture analyzer showed no signicant differences after one-day storage (Fig. 1B). Elasticity was the most stable
parameter during the 5 day storage. Values did not signicantly
differ among breads. Moreover, chewiness and hardness showed
signicant differences (p 0.007 and 0.001, respectively) after the
second day of storage. Cohesiveness was the other textural

M.A. Lazo-V
elez et al. / Journal of Cereal Science 64 (2015) 63e69

Table 5
Overall scores of soybean-fortied breads compared to wheat bread estimated by
nutritional attributes, dough properties, baking, crumb texture and organoleptic
tests.
Parameters

Control

Flour enriched witha

Wheat our SBF-1 SBF-2 SBF-3 SBF-4 SBC

Nutritional
Protein content
Amino acid composition
Dough machinability
Alveograph P
Mixing tolerance index
Baking performance
Water absorption
Dough mixing time
Bread volume
App. bread density
Acceptability
Overall acceptability
Texture after 5 days storageb
Desirability value

1
1

1
1

1
1

1
1

1
1

1
1

1
1

1
0.5

1
1

1
1

1
0

1
0.5

1
1
1
1

1
1
2
1.5

2
1
2
2

1
1.5
1.5
1

1.5
1.5
1.5
1.5

3
1
1
1

1
1
1

0
0.5
1.5

0
1
4

0
1.5
0.5

0
1
1

0
1
1.5

The experimental SB enriched ours were produced by substitution of 6.0% SBF1; 6.1% SBF-2; 6.3% SBF-3, 6.2% SBF-4 and 4.5% SBC of the rened bread our.
b
Average of cohesivity, hardness and chewiness.

parameter in which all sample values were very close. The control
and SBC enriched breads showed the higher and lower values
respectively (Fig. 1B).
Ivanovski et al. (2012) showed that SB our or protein isolates
that substituted wheat our 20 or 12%, respectively, also inuenced
both bread avor and texture. SB enriched breads had a signicantly stronger avor characterized as beany. Mahmoodi et al.
(2014) described that, appearance, avor and taste in breads containing 3, 7 or 12% defatted SB decreased with higher substitution
levels. These authors concluded that the best formulations were
attained between 3 and 7% substitution.
Finally, overall evaluations of the best performing ours were
made (Table 5). Scores for white bread compared to the different SB
sources were obtained according to the improvement in protein
quality, changes in rheological properties, features of breads and
organoleptic tests. The best scores were for SBC followed by the
SBF-3 whereas the worst treatment was the composite our containing SBF-2 mainly because its addition worsened rheological
properties, lowered bread volume and increased apparent density
of bread.

4. Conclusions
In conclusion, the SBC and SBF-3 blended with rened wheat
our produced the best enriched breads in terms of organoleptic
perception, texture, and bread properties. These SB proteins yielded
slightly higher amounts of bread due to their higher dough water
absorption. The SB enriched breads had improved protein concentration and amounts of the limiting amino acid lysine and
consequently a better protein quality as has been documented by
other investigations. The SB-enriched breads had comparable
sensory acceptability values and texture throughout 5 days storage
compared to the control wheat bread. Additionally, correlation
between functional soybean properties, baking properties of pup
loaves, and rheological properties of SB composite ours (farinograms and alveograms) evidenced that the functional characteristic
of the SB ours especially in terms of PDI and NSI should be taken
into consideration for the production of fortied yeast-leavened
breads.

69

Acknowledgments
The authors would like to acknowledge USSEC for providing the
array of soybean samples and sponsoring this research. In addition,
authors acknowledge the continuous support of the Consejo

xico, Secretara
Nacional de Ciencia y Tecnologa (CONACyT), Me

n
Nacional de Estudios Superiores, Ciencia, Tecnologa e Innovacio
(SENESCyT), Ecuador and the Nutrigenomic Research Chair of Tec
gico de Monterrey for providing research and support funds for
nolo
the senior author.
Appendix A. Supplementary data
Supplementary data related to this article can be found at http://
dx.doi.org/10.1016/j.jcs.2015.04.007.
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