Food Bioscience 8 (2014) 33 –36

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Processing and bread-making potential of proteins

isolated from malted and non-malted pea seeds
by ultrafiltration/diafiltration

Sébastien Villeneuvea,b, Martin Mondora,b,n

a
Agriculture and Agri-Food Canada, Food Research and Development Centre, 3600 Casavant Blvd West,
Saint-Hyacinthe, QC, Canada J2S 8E3
b
Institute of Nutrition and Functional Foods (INAF), Laval University, QC, Canada G1K 7P4

art i cle i nfo ab st rac t

Article history: The bread-making potential of a pea protein isolate produced by an ultrafiltration/
Received 4 June 2014 diafiltration (UF/DF) process applied to malted seed has not been studied, to date. Thus,
Received in revised form the objective of this work was to study the impact of supplementing breads, at a 10% level,
9 October 2014 with malted pea protein isolate produced by an UF/DF processes with a 50 kDa hollow
Accepted 20 October 2014 fibers membrane. Malted pea protein isolate with approximately 90% of protein (dry basis)
was obtained. Flour substituted with this isolate showed good bread-making potential

Keywords: allowing the production of bread with protein content over 20%, which was comparable to

Yellow peas the protein content obtained for bread produced from non-malted pea protein isolate. The

Protein isolate bread produced from malted pea protein isolate had a specific volume of 3.970.1 cm3 g
1

Germination which was slightly lower than for the one produced from non-malted pea protein isolate

Malting (4.470.1 cm3 g
1) but comparable to the control bread produced from 100% refined bread

Ultrafiltration/diafiltration flour (4.270.1 cm3 g
1). Color (Ln,an,bn) of the crust and of the crumb of breads produced

Bread from UF/DF isolates was barely affected by the malting process but slightly differed from
the color of bread produced from 100% refined bread flour.
Crown Copyright & 2014 Published by Elsevier Ltd. All rights reserved.

1. Introduction level of substitution. Generally, the substitution of wheat

Fortifying wheat breads with pea flours or isolates to improve
their nutritional value has received considerable interest
(Morad, Leung, Hsu, & Finney, 1980; Hsu, Leung, Finney, &
Morad, 1980; Hsu, Leung, Morad, Finney, & Leung, 1982;
Sadowska, Blaszczak, Fornal, Vidal-Valverde, & Frias, 2003;
Des Marchais, Foisy, Mercier, Villeneuve, & Mondor, 2011;
Mondor, Guévremont, & Villeneuve, 2014). The impact of
fortification is dependent on the pea processing and on the
flour with pea-based ingredients at a level of more than 10%
is harmful to the processing of bread (Sadowska et al., 2003).
A process that has been investigated in past studies is the
germination (or malting) of peas prior to the preparation of
pea flours (Sadowska et al., 2003; Mondor et al., 2014) or pea
isolates (Hsu et al., 1982). It has long been recognized that
germination generally enhances the nutritive value of seeds
(Hübner and Arendt, 2013) and would thus result in flours/
isolates with improved nutritional value. It was also

n
Corresponding author at: Agriculture and Agri-Food Canada, Food Research and Development Centre, 3600 Casavant Blvd West,
Saint-Hyacinthe, QC, Canada J2S 8E3. Tel.: þ1 450 768 3297; fax: þ1 450 773 8461.
E-mail address: martin.mondor@agr.gc.ca (M. Mondor).

http://dx.doi.org/10.1016/j.fbio.2014.10.004
2212-4292/Crown Copyright & 2014 Published by Elsevier Ltd. All rights reserved.
34 Food Bioscience 8 (2014) 33 –36
demonstrated that germinated or malted pea flours can be
successfully incorporated into breads, at a level of 10%,
without major defects to the bread structure (Sadowska
et al., 2003; Mondor et al., 2014). However, in the two afore-
mentioned studies, the improvement of the breads in terms of
protein content remained limited. On the other hand, the
incorporation of pea protein isolate produced from germi-
nated pea (and from non-germinated pea) into breads, at a
level of 8%, had negative effects on bread volume (Hsu et al.,
1982). However, no significant difference was observed in
terms of loaf volume between the bread substituted with
germinated pea protein isolate and the one substituted with
non-germinated pea protein isolate, indicating that the detri-
mental effect of substitution is not attributable to the germi-
nation process but to the processing of peas into protein
isolates (Hsu et al., 1982).
It is well known that the production of plant protein
isolates at industrial scale by isoelectric precipitation, and
as it was the case in the study of Hsu et al. (1982), results in
protein denaturation due to the use of acids during the
precipitation step. This will in turn negatively impact the
functional properties of the protein isolate (Petruccelli &
Anon, 1994; Wagner, Sorgentini, & Anon 2000; Taherian
et al., 2011). One possible explanation for the detrimental
effect observed by Hsu et al. (1982), on the loaf volume, when
substituting breads at 8% level with pea protein isolates is
that the denatured proteins would be less prone to take part
in the formation of bread structure, which would negatively
impact its specific volume.
In a recent work, it was demonstrated that pea protein
isolates produced by an ultrafiltration/diafiltration (UF/DF)
process have significantly better functional properties than
commercial pea protein isolates produced by isoelectric pre-
cipitation (Taherian et al., 2011). The good bread-making
potential of this isolate, at a level of 10%, was also confirmed
(Des Marchais et al., 2011). To our knowledge, the bread-
making potential of pea protein isolates produced by an
UF/DF processes applied to malted seed has not been studied
to date. Nevertheless, even at a substitution level of 10%, it
would allow the production of breads with improved nutri-
tional value and with protein content significantly higher
than when flours are used for substitution. Thus, the objec-
tive of this work was to study the impact of supplementing
breads, at a 10% level, with pea protein isolate produced by an
UF/DF processes from malted yellow peas.
2. Materials and methods
2.1. Raw materials
Refined bread flour (Robin Hood KEYNOTE 80 Bleached
Enriched Flour; Horizon Milling, Montreal, Quebec, Canada)
was used. Certified #1 Eclipse yellow peas were purchased
from Wagon Wheel Seed Corporation (Churchbridge, SK,
Canada). Commercial pea protein isolate was obtained from
Nutri-pea Limited (Portage La Prairie, MB, Canada) and used
as control.
2.2. Malting process and production of pea protein isolates
by UF/DF
The malting procedure previously described in Mondor et al.
(2014) was used in this study. The brittle acrospires of dry peas
were removed by sieving and the peas were processed into
flours as previously described (Mondor et al., 2014). Then,
isolates were produced by extracting pea flours produced from
non-malted and malted yellow peas, in water with 1:15 w/w
solid/liquid ratio at room temperature and pH 7.5 for 1.25 h.
Extracts were purified by UF/DF processes performed with a
50 kDa hollow fibers membrane, by applying a volume con-
centration ratio of 5, in order to obtain protein isolates, as
previously described by Taherian et al. (2011). A 50 kDa
membrane was selected based on the work of Fuhrmeister
and Meuser (2003) who demonstrated that the most favourable
combination of permeate flux and pea protein yield in eco-
nomic terms is achieved at that cut-off. A 50 kDa membrane
was also used by Frederickson, Biot, Alminger, Carlsson, and
Sendberg (2001) for the production of high quality pea protein
isolate. The resulting pea protein isolates were lyophilized and
placed in aluminium pouches which were hermetically sealed
and stored at 4 1C until used. Ash and protein contents of the
flours and isolates were measured based on the methods
provided by Taherian et al. (2011).
2.3. Bread preparation
Bread was prepared according to AACC International Method
10–10.03 applying the 100 g-flour procedure. Water absorption
of 62.5% and mixing time of 7 min were applied. Three different
breads, such as bread made using pea protein isolates produced
by UF/DF from non-malted peas (NMPB), bread made using pea
protein isolates produced by UF/DF from malted peas (MPB) and
bread made using commercial pea protein isolates (CPB), were
prepared in duplicate with refined bread flour substituted at a
10% level (dry basis) with the pea protein isolates. Control bread
was also made using 100% refined bread flour. After the cooling
period (1 h after removal from the oven), the breads were
weighed and their volume was determined by rapeseed dis-
placement (National Mfg, Co., Lincoln, Nebraska, U.S.A.). The
specific volume was then calculated by dividing the volume of
the bread by its mass. Color measurements of crust and crumb
were performed using a Minolta colorimeter model CR-300
(Konica Minolta Sensing Americas, Ramsey, NJ, U.S.A.). Mea-
surements were done in a light testing box (Macbeth, The Judge
II, Grand Rapids, MI, USA) under “day” light intensity. Results
are expressed in terms of the Hunter colour system Ln (black-
ness to whiteness), an (greenness to redness) and bn (blueness to
yellowness). Bread crumb was dried in a vacuum oven at 92 1C
for 16 h and protein content was determined using a Kjeldahl
digestion system (Tecator, Höganäs, Sweden) and a conversion
factor of N  6.25 for peas and N  5.7 for refined wheat flour.
2.4. Statistical analysis
Analysis of variance was performed a priori on each para-
meter using SAS software (version 8.2, SAS Institute Inc. Cary,
NC, USA). Multiple comparison procedure (Least Significant
 Food Bioscience 8 (2014) 33 –36 35

Difference, p¼ 0.05) was performed a posteriori to compare
parameters whose variance was significantly different.
3. Results and discussion
3.1. Composition of yellow pea protein isolates
Table 1 presents the composition of isolates processed from
non-malted and from malted peas by UF/DF, as well as the
one of the commercial pea protein isolate. Both pea protein
isolates produced by UF/DF from the non-malted and malted
pea flours resulted in products with similar protein and ash
contents. This is a good indication that UF/DF processes
performed with a 50 kDa hollow fibers membrane, as pre-
viously described by Taherian et al. (2011) are appropriate to
produce pea protein isolates from both non-malted and
malted peas. The commercial pea protein isolate has a
protein content of 88.770.2% (dry basis) and an ash content
of 5.270.0%.
3.2. Bread characteristics
Bread characteristics in terms of protein content, specific
volume and colour are presented in Table 1. Substitution with
pea protein isolates allowed to produce breads with protein
content over 20% dry basis (21.573.3% for NMPB, 21.870.9%
for MPB and 20.470.1% for CPB) compared to 14.170.3% for
refined wheat flour bread. A protein content of this magni-
tude is relatively high considering that reports in the litera-
ture conclude that high protein breads contain up to 15–20%
protein (Mohamed, Rayas-Duarte, Shogren, & Sessa, 2006).
In terms of specific volume, the NMPB showed a higher
specific volume than for the MPB and the CPB (4.470.1 cm3
g
1, 3.970.1 cm3 g
1 and 3.070.2 cm3 g
1, respectively). The
specific volume measured for the NMPB and MPB are similar
to the value measured for refined wheat flour bread
(4.270.1 cm3 g
1). On the other hand, the value measured
for the CPB is significantly lower, which is in agreement with
the observations of Hsu et al. (1982) that the substitution of
yellow pea protein isolates produced by isoelectric precipita-
tion, in a bread formulation, has a negative impact on bread
specific volume. The difference observed in the specific
volume between the NMPB and MPB is possibly due to the
presence of amylases in the pea protein isolate produced
from malted peas. Pea amylases have a molecular weight in
the order of 43–44 kDa (Beers & Duke, 1990) which is close to
the nominal molecular weight cut-off of the UF membrane.
This indicates that the amylases activated during the malting
process and not inactivated by the drying step at 22 1C are
partially retained and are thus present in the malted pea
protein isolate. Moreover, presence of amylases in the isolate
could at the end impact the dough rheological properties.
According to Mondor et al. (2014), heat treating malted yellow
pea flour at 80 1C in order to inactivate amylases led to dough
mixing properties similar to those of non-malted yellow pea
flour. On the other hand, low specific volume observed for the
CPB would be mainly due to the denaturation of the proteins
present in the isolate. Pea proteins in their native state would
interact more directly with wheat proteins while denatured
proteins from the commercial isolate would alter dough
properties more indirectly because of water competition
and gluten dilution, similar to what was observed in the case
of wheat-soy proteins interactions (Ribotta, Edel Leon, Pérez,
& Anon, 2005; Roccia, Ribotta, Pérez, & Edel Leon, 2009) and
with durum wheat semolina- pea proteins (Mercier et al.,
2012).
In terms of colors, the crust darkening of the breads
containing both malted and non-malted UF/DF pea protein
isolates as compared to control bread might have been
attributed to an increased Maillard reaction taking place
during baking of the loaves, due to the higher protein content
of enriched breads. In the Maillard reaction reducing carbo-
hydrates react with free amino acid side chains of proteins
and lead to amino acid sugar reaction products (polymerised
protein and brown pigments) (Hallén, Ibanoglu, & Ainsworth,

Table 1 – Characterization of pea protein isolates and breadsn.

Parameters 100% Refined bread flour (Control) Commercial isolate Non-malted (UF/DF) Malted (UF/DF)

Isolates
Protein content (g 100 g- – 88.770.2a 91.571.9b 89.373.9ab
dry matter
1)
Ash (g 100 g-dry matter
1) – 5.270.0b 3.870.4a 3.570.6a

Breads
Protein content (g 100 g- 14.170.3a 20.470.1b 21.573.3b 21.870.9b
dry matter
1)
Specific volume (cm3 g
1) 4.270.1bc 3.070.2a 4.470.1c 3.970.1b

Colour of the crust

Ln value 48.771.3c 61.472.1d 35.970.1a 38.373.6b
an value 14.070.1b 13.171.1a 13.870.2b 15.370.8c
bn value 34.271.2c 36.070.5d 19.970.1a 24.576.7b

Colour of the crumb

Ln value 86.470.2d 80.770.5a 83.270.8b 83.570.8c
an value
5.370.0a 1.070.2d
4.070.0b
3.770.1c
bn value 17.070.2a 20.970.6b 20.870.6b 20.970.6b

n
Values not sharing a common letter on the same row are significantly different (po0.05).
36 Food Bioscience 8 (2014) 33 –36
2004). On the other hand, the crust of the bread containing
the commercial isolate showed a brighter color than the
control bread. A possible explanation might be that the
denatured proteins are less prone to take part to Maillard
reactions. However, this remains to be experimentally con-
firmed. Although some significant color differences were
instrumentally observed for the crumb, differences based on
visual observation may not be that discernible. Our observa-
tions are in agreement with those of Charoenthaikij et al.
(2010) for germinated brown rice.
4. Conclusion
Combination of peas malting and UF/DF process performed
with a 50 kDa hollow fibers membrane is an appropriate
approach to produce pea protein isolates from malted peas.
Flour substituted with pea protein isolates produced from
malted yellow peas showed good bread making potential,
when compared to flour substituted with commercial pea
protein isolate.
Acknowledgements
The authors would like to acknowledge the technical assis-
tance of Claire Baudin, Louis-Philippe Des Marchais, Hélène
Drolet and Michel Gros-Louis.
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