J Food Sci Technol

https://doi.org/10.1007/s13197-019-03785-8

ORIGINAL ARTICLE

Effect of pH on protein extraction from sour cherry kernels

and functional properties of resulting protein concentrate
Mehtap Çelik1 • Melih Güzel2 • Metin Yildirim3

Revised: 26 March 2019 / Accepted: 12 April 2019

Ó Association of Food Scientists & Technologists (India) 2019

Abstract The aims of this research were to examine the Introduction

effect of pH on extraction of proteins from sour cherry
(Prunus cerasus L.) kernels, and to investigate the func- Because of the growing consumers’ concerns, the food
tional properties of the resulting protein concentrate. The industry is persistently searching for healthier and inex-
optimum pH values for the protein extraction and iso- pensive protein ingredients to substitute those originated
electric precipitation were determined as 10.0 and 4.5, from animal sources (e.g., gelatine, casein, whey proteins,
respectively. The protein concentrate contained and ovalbumin). Therefore, research on the plant proteins
4.03 ± 0.16% moisture, 3.31 ± 0.17% ash, 2.94 ± 0.36% has become a chief issue to be addressed (Adebowale et al.
carbohydrate, 1.93 ± 0.16% lipid, and 80.48 ± 2.38% 2011).
protein. Water holding capacity, oil holding capacity and Cherry belongs to the genus Prunus of the Rosaceae
the least gelling concentration of the protein concentrate family. Sour (tart) cherry (Prunus cerasus L.) has a higher
were 2.42 ± 0.09 g water/g, 1.73 ± 0.17 g oil/g and 8%, acid/sugar ratio than sweet cherry (Prunus avium L.).
respectively. Results showed that emulsifying activity and Therefore, sour cherry is mostly used for the production of
stability indices, foaming capacity and stability of protein juice, concentrate, jam, puree, marmalade or pie filling
concentrate were 38.91 ± 2.50 m2/g, 37.49 ± 2.41 min, while sweet cherry is generally consumed fresh (Toydemir
35.00 ± 3.54% and 71.80 ± 7.25% (after 30 min), et al. 2013; Yılmaz et al. 2018). Worldwide total sour
respectively. The functional and chemical properties of the cherry production is about 1.38 million tons in 2016
protein concentrate indicate that it may find application as (FAOSTAT 2017), approximately 85% of which is pro-
functional ingredient for various food products. cessed into various food products (Toydemir et al. 2013),
creating a high amount of seed as a waste material that
Keywords Sour cherry kernel  Protein concentrate  causes an important disposal issue for the food industry
Chemical properties  Functional properties (Yılmaz and Gökmen 2013). Currently, large amounts of
seeds are discarded at the processing plants. For example,
15,000–20,000 tons of sour cherry seeds were obtained in
the United States for the year 2014, mainly from juice
processing factories (Korlesky et al. 2016). The absence of
& Metin Yildirim alternatives for reusing these wastes makes further inves-
[email protected] tigations really important. Sour cherry kernels comprise
1
Department of Food Engineering, Faculty of Engineering,
25.3–31.7% protein, 9.5–30.3% dietary fibre and
Hitit University, 19030 Çorum, Turkey 17.0–41.9% oil (Korlesky et al. 2016; Yılmaz and Gökmen
2 2013; Bak et al. 2010; Kamel and Kakuda 1992; Lazos
Department of Food Processing, Şiran Mustafa Beyaz
Vocational School, Gümüşhane University, 1991). Garcı́a et al. (2015) has also showed the presence of
29700 Gümüşhane, Turkey antioxidant and antihypertensive peptides in sour cherry
3
Department of Food Engineering, Faculty of Engineering, kernel proteins.
Niğde Ömer Halisdemir University, 51240 Niğde, Turkey

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Functional properties of proteins are those physico- (InoLab WTW pH 720, Weilheim, Germany). The result-
chemical properties which determine their behaviour in ing slurry was stirred for 150 min at room temperature
food systems during preparation, processing, and storage. meanwhile the pH of the slurry was kept constant by
Solubility, emulsifying, foaming, water and oil holding, readjusting every 30 min, if needed. Then, the slurry was
and gelation are a few examples of important functional filtered through a coarse filter paper (Whatman Grade 1)
properties of proteins. Therefore, proteins are used as a and the filtrate was collected and analysed for protein
functional ingredient in food products to form certain content to calculate the protein extraction yield (the
sensory characteristics and/or to improve nutritional qual- amount of protein in supernatant*100/the amount of pro-
ity (Kinsella 1981; Damodaran 1997). Research on the tein in the sample) and to determine optimum extraction
functional properties of food proteins obtained from dif- pH. After that, the pH of the filtrate was adjusted to 4.5
ferent sources is an essential in order to understand better with 2 N HCl, allowed to stand for 15 min, and further
their roles in food systems. filtered through Whatman Grade 1 filter paper. The pre-
The sour cherry kernel could be a valuable source of cipitated proteins were re-suspended in distilled water and
proteins for use as functional food ingredients. At present, its pH was adjusted to 7.0 and then dried at 50 °C for
to the best of the researchers’ knowledge, no information 12–18 h with an air flow oven (Memmert 100-800, Sch-
on the functional properties of these proteins is available. wabach, Germany) and stored at - 18 °C until use.
Thus, extracting the proteins from sour cherry kernels and
understanding the functional properties of the resulting Proximate composition
protein concentrate is crucial for food use. Therefore, the
specific objectives of the present study were to: (1) The dry matter and ash contents of sour cherry kernel flour,
investigate the extractability of sour cherry kernel proteins defatted flour and protein concentrate were determined by
from defatted meal as a function of pH, and (2) determine gravimetric method (AOAC 1997). Ankom extractor
the selected functional properties and chemical composi- (Ankom XT10 Extractor, Macedon, NY, USA) was used to
tions of the resulting protein concentrate. analyse the lipid content. The total carbohydrate content of
the samples was determined according to the phenol sul-
phuric acid method (Geater and Fehr 2000). The micro-
Materials and methods Kjeldahl method was used to analyse the nitrogen content
of the samples (AOAC 1997). The value of 6.25 was used
Materials as protein conversion factor.

The sour cherry kernels were obtained from a local com- Colour parameters
pany in the city of Tokat, Turkey. Samples were stored in
plastic bags at 4 °C until use. Sodium caseinate containing The CIE Lab parameters (L*, a*, b*) of the sour cherry
13.5–16.0% nitrogen was purchased from Sigma-Aldrich kernel flour, defatted flour and protein concentrate which
(St. Louis, MO, USA). All the chemicals and reagents used were spread with a thickness of 1 cm over glass petri
were of analytical grade and used without further dishes were directly read from 3 different points via a
treatment. colorimeter (Minolta, CR-300, Osaka, Japan) calibrated by
means of a white tile (L* = 96.97, a* = 0.16, b* = 1.86) as
Extraction of proteins reference. In this coordinate system, the a* value varies
from green (-) to red (?), the b* value varies from blue
Sour cherry kernel protein concentrate (SCKPC) was pre- (-) to yellow (?) and the L* value is a measure of light-
pared by alkaline extraction followed by isoelectric pre- ness, ranging from 0 (black) to 100 (white).
cipitation. The kernels were ground (B 1 mm) with a
coffee grinder (Bosch MKM 600, Munich, Germany). Sodium dodecyl sulphate–polyacrylamide gel
Then, the sour cherry kernel flour (SCKF) was defatted electrophoresis (SDS-PAGE)
four times (1 h each) using n-hexane in a 1:6 flour to
solvent ratio at room temperature. The defatted sour cherry The electrophoretic profiles of the proteins from DSCKF
kernel flour (DSCKF) was left in a fume hood for 12 h to and SCKPC were determined according to the method
remove hexane residues, and then transferred to a sealed described by Laemmli (1970). The sample equivalent to
glass jar and stored at room temperature until use. For 5 mg of protein was dissolved in 1 mL of sample buffer
protein extraction, the DSCKF was dispersed in distilled [3.8 mL of distilled water, 1 mL of 0.5 M Tris–HCl pH 6.8
water (5%, w/v) and the pH of the slurry was adjusted to buffer, 0.80 mL of glycerol, 1.6 mL of 10% (w/v) SDS,
1.0–12.0 with 2 N HCl or 2 N NaOH using a pH meter 0.8 mL of b-mercaptoethanol, 0.4 mL of 0.05% (w/v)

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bromophenol blue] and then heated at 95 °C for 5 min. Foaming capacity and foaming stability
After cooling, a 10 lL aliquot was loaded to 1 mm thick
gels (4% stacking; 12% separating). A mixture of standard The foaming capacity and foaming stability of SCKPC
proteins (6.5–200 kDa, catalogue number S8445, Sigma- were determined using the method of Moure et al. (2001)
Aldrich, St. Louis, MO, USA) was used as molecular with slight modifications. 0.5 g SCKPC was dispersed in
weight marker and the gels were stained with Coomassie distilled water, the pH was adjusted to 7.0, and the volume
Brilliant Blue G-250 and de-stained with 10% acetic acid. was made up to 40 mL (1.25%, w/v) with distilled water.
For stacking gel 25 mA and for separating gel 35 mA The dispersion was homogenized with Ultra Turrax (T18
currents (Consort E815, Turnhout, Belgium) were basic, IKA-Werke GmbH & Co. KG, Staufen, Germany) at
employed. 20,000 rpm for 2 min at room temperature and then
instantly transferred to a measuring cylinder, and the total
Protein solubility volume and volume of liquid phase were recorded.
Foaming capacity was calculated as:
Protein solubility of the sour cherry kernel protein con- ðV1  V2 Þ  100
centrate was determined as described by Beuchat et al. Foaming capacity ð%Þ ¼
V2
(1975) with slight modifications. Dispersions containing
5% (w/v) SCKPC were prepared and the pH of dispersions where V1 was the total volume after homogenization, V2
was adjusted to 1.0–12.0 using either 1 N HCl or 1 N was the total volume before homogenization.
NaOH. The dispersions were stirred at room temperature The foaming stability was calculated by the following
for 90 min after initial pH adjustment. The pH was checked equation by measuring the change in foam volume after 0,
at 30 and 60 min, and if needed readjusted to the stipulated 10, 30, 60, 90 and 120 min of storage.
value. Then, the dispersions were centrifuged at 40009g Vt  100
for 30 min. Protein content in the supernatant was deter- Foaming stability ð%Þ ¼
Vk
mined by micro-Kjeldahl method (AOAC 1997) and the
where Vt was the volume of foam at time t, Vk was the
protein solubility was calculated as:
foam volume at 0 min after homogenization.
W1  100
Protein solubility ð%Þ ¼
W0 Emulsifying activity and stability indices
where W1 was the amount of protein in the supernatant (g),
W0 was the amount of protein in the sample (g). Emulsifying activity (EAI) and emulsifying stability indi-
ces (ESI) were determined according to the method of
Water-holding capacity (WHC) and oil holding Pearce and Kinsella (1978). For emulsion formation,
capacity (OHC) 6.6 mL of commercial sunflower oil was added to 20 mL
of SCKPC dispersions (0.1% protein, w/v, pH 7.0) and
Water and oil holding capacities of SCKPC were deter- homogenized by using Ultra Turrax, (T18 basic, IKA-
mined by using the method outlined by Naczk et al. (1985). Werke GmbH & Co. KG, Staufen, Germany) at
For the water holding capacity, 0.5 g SCKPC was dis- 20,000 rpm for 1 min. Fifty microliters of emulsion (by
persed in 4 mL of distilled water in a test tube. The dis- avoiding the foam layer) were removed carefully from the
persion (pH 7.0) was stirred for 30 s every 10 min and held emulsions, immediately mixed with 4.95 mL of 0.1% (w/v)
up for 70 min, and then centrifuged at room temperature SDS solution (1:100 dilution) and vortexed for 10 s, and
for 15 min at 20009g. The supernatant was drained for the absorbance of the mixture (A0) was measured at
10 min at 45° angle. The gain in weight was recorded as 500 nm versus 0.1% SDS as blank using a spectropho-
WHC (g water/g sample). tometer (Perkin Elmer UV/Vis spectrophotometer, Lambda
In the determination of oil holding capacity, 0.5 g EZ 201, Waltham, MA, USA). Ten minutes later, another
SCKPC was dispersed in 3 mL of commercial sunflower 50 lL of emulsion were removed from the emulsions as
oil in a test tube. The dispersion was stirred for 30 s every mentioned above, immediately mixed with 4.95 mL of
5 min and allowed to stand for 30 min, and then cen- 0.1% SDS solution and vortexed for 10 s, and the absor-
trifuged at room temperature for 25 min at 16009g. The bance of the mixture (A10) was determined at 500 nm. EAI
supernatant was drained for 5 min at 45° angle. The gain in and ESI were calculated using the following equations:
weight was recorded as oil holding capacity (g oil/g sam-
2  2:303  A0  N
EAI m2 =g ¼
ple). Sodium caseinate was used as a reference for WHC c  u  10; 000
and OHC tests.

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A0  t Table 1 Proximate composition of sour cherry kernel flour (SCKF),

ESIðminÞ ¼ defatted sour cherry kernel flour (DSCKF), and sour cherry kernel
A0  A10
protein concentrate (SCKPC)
where, A0 was the absorbance of the diluted emulsion Parameters (%) SCKF DSCKF SCKPC
immediately after homogenization, N was the dilution
factor (100), c was the protein concentration of protein Moisture 4.64 ± 0.04 6.94 ± 0.35 4.03 ± 0.16
dispersion (0.001 g/mL), u was the oil volume fraction of Protein 32.21 ± 0.83 42.07 ± 0.09 80.48 ± 2.38
emulsion (6.6/26.6 = 0.248), A10 was the absorbance at Lipid 34.75 ± 0.68 9.00 ± 0.28 1.93 ± 0.16
10 min after homogenization, t was the time interval, Total carbohydrate 10.16 ± 0.73 16.28 ± 1.63 2.94 ± 0.36
10 min. Ash ND ND 3.31 ± 0.17
All the data are expressed as mean ± standard deviations and are the
The least gelation concentration mean of three replicates (n)
ND Not determined
The least gelling concentration (LGC) of the samples was
measured according to the method described by Coffman
and Garcia (1977). 5 mL of aqueous dispersions (2–14%
w/v, pH 7.0) of SCKPC were placed in a boiling water bath could be the insufficient solvation of polar lipids such as
for 1 h, followed by rapid cooling to 4 °C in an ice bath, phospholipids and free fatty acids by hexane (Makri et al.
and then held up for 2 h. Gel formation was evaluated by 2005), the extraction of lipids without drying or roasting
inverting the tubes containing the treated dispersions. The the kernel and performing the extraction at room temper-
least gelation concentration was then determined as the ature. Higher extraction efficiency was reported for broad
concentration at which the sample from the inverted tube bean (Fernandez-Quintela et al. 1997), hyacinth bean
did not fall or slip. (Subagio 2006), and chickpea (Kaur and Singh 2007). The
removal of lipid caused an increase in protein and total
Statistical analysis carbohydrate content of defatted sour cherry kernel flour
(Table 1).
Three independent determinations were used to obtain
mean values and standard deviations. One-way analysis of Extraction conditions of proteins
variance and Duncan tests were used for the statistical
evaluation and comparison of the data in the Minitab There are various techniques such as ultrafiltration or
program (Minitab release 12.1, 1998, Minitab Inc., State reverse osmosis, extraction with organic solvents or neutral
College, PA, USA) at p \ 0.05 significance level. salts for protein isolation, but the extraction in alkaline
conditions followed by isoelectric precipitation is more
advantageous due to cheaper chemicals used and simpler
Results and discussion equipment needed. Therefore, the alkaline extraction was
chosen in this study. In order to determine the optimum pH
Proximate composition for the protein extraction from sour cherry kernel, DSCKF
samples were dispersed in distilled water (5% w/v), and the
Moisture, protein, lipid, and total carbohydrate contents of pH of the dispersions were adjusted from 1.0 to 12.0. After
SCKF and DSCKF are shown in Table 1. The proximate mixing and filtration, the yield of protein extraction was
composition of SCKF was similar to those reported in the determined (Fig. 1). The yield of protein extraction from
literature. The moisture content of sour cherry kernel was sour cherry kernel was high at pH 1.0 (75.3 ± 4.60%), 2.0
recorded as 3.91% (Yılmaz and Gökmen 2013). The pro- (78.9 ± 4.80%), 9.0 (67.8 ± 2.60%), 10.0 (74.1 ± 2.50%),
tein content of sour cherry kernel was found to be 25.3% by 11.0 (70.5 ± 0.60) and 12.0 (76.9 ± 0.70), and there were
Lazos (1991) and 37.80% by Kamel and Kakuda (1992). no statistically significant differences (p [ 0.05) among
The reported lipid content of sour cherry kernel was 26.0% these extraction pHs. The yield of protein extraction was
(Lazos 1991), 32.0–36.0% (Bak et al. 2010), 41.9% (Kamel minimal at pH 4.0 (31.9 ± 2.30%); therefore, the isoelectric
and Kakuda 1992). Lazos (1991) and Yılmaz and Gökmen point of the sour cherry kernel proteins was assumed to be
(2013) reported that the total carbohydrate content of sour pH 4.0. However, there were no significant differences
cherry kernel was 34.5% and 46.6%, respectively. (p [ 0.05) among the extraction pHs 4.0 (31.9 ± 2.30%),
After extraction, the lipid content of the kernel flour was 5.0 (34.9 ± 1.16%), and 6.0 (38.7 ± 2.71%). The proteins
reduced from 34.75 ± 0.68% to 9.00 ± 0.28%, indicating extracted at these pHs are mainly proteins with higher iso-
lower lipid extraction efficiency. The reason for this result electric point (Makri et al. 2005) and small polypeptides

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Fig. 1 Effect of pH on protein 100

extraction from defatted sour
cherry kernel flour 90

Protein extraction yield (%)

80
70
60
50
40
30
20
10
0
1 2 3 4 5 6 7 8 9 10 11 12
pH

(Subagio 2006). The extraction pHs of too high or too low (2006) reported a higher yield of 37–40% for hyacinth bean
may result in a structural damage in proteins; therefore, protein isolate.
although a higher yield of protein extraction was obtained at
pH 12.0 (76.9%), the optimum pH value for protein SDS-PAGE
extraction was assumed to be pH 10.0.
SDS-PAGE analysis was performed to determine apparent
Production and composition of SCKPC molecular weight distribution of the proteins recovered in
SCKPC and the electrophoretogram generated is presented
Sour cherry kernel protein concentrate was produced by in Fig. 2. As can be seen from the electrophoretogram,
extracting the proteins at pH 10.0 and precipitating at pH apparent molecular weights of proteins in SCKPC varied
4.5 at room temperature. While the minimum solubility from 14 to 66 kDa under reducing and denaturing condi-
was observed at pH 4.0, the isoelectric precipitation was tions. More specifically, five major protein bands (sub-
performed at pH 4.5 due to the formation of a firmer pre- units) with approximate apparent molecular weights of 45,
cipitate. The protein rich product obtained by this way was 36, 29, 23, and 16 kDa were observed.
regarded as protein concentrate because its protein content Almost all of the proteins present in DSCKF were also
in the dry matter (83.86%) was less than 90%. The prox- present in SCKPC, indicating that practically all the kernel
imate composition of SCKPC (Table 1), except for protein protein sub-units were transferred to SCKPC. Additionally,
content was parallel to the results reported for barley pro- the electrophoretogram showed that no significant protein
tein concentrate (Fernandez-Quintela et al. 1997) and degradation was occurred during protein concentrate pro-
chickpea protein concentrate (Ghribi et al. 2015). duction. However, some of the minor bands between
While 74.1 ± 2.50% of the sour cherry kernel proteins 29–36 kDa in DSCKF were absent in SCKPC. This may be
were solubilized at pH 10.0, only 35.56 ± 0.52% of the due to the fact that these protein bands were not extracted
proteins were recovered in SCKPC by precipitating at pH at pH 10.0 or remain soluble at pH 4.5.
4.5, indicating a poor protein recovery rate (18.59 g of
SCKPC was produced from 100 g of DSCKF). In order to Colour properties
increase the protein recovery rate, higher extraction tem-
peratures, multi-stage extraction and membrane filtration Among the colour parameters (CIE L*, a* and b*), the L*
techniques can be used. Additionally, enzymes such as and b* values of SCKF (69.73 ± 1.08 and 27.73 ± 0.15),
pectinase, cellulases, phytase and xylanase can assist to DSCKF (84.19 ± 1.94 and 15.80 ± 0.71) and SCKPC
increase the extractability of proteins bound to cellular (55.43 ± 0.96 and 23.71 ± 0.50) were considerably dif-
components and phytate. Studies on the protein extraction ferent from each other (p \ 0.05); however, there were no
from different sources showed that the recovery rates were significant differences between a* values of SCKF
varied significantly. While Kaushik et al. (2016) found a (5.11 ± 0.18) and SCKPC (5.67 ± 0.21) (p [ 0.05).
yield of 20.09% for flaxseed protein concentrate; Subagio DSCKF had the highest L* value and the lowest

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Additionally, information on the solubility of proteins is

important for extracting and purifying proteins from natu-
ral resources. The solubility of a protein depends on such
factors as pH, ionic strength, temperature, concentration
and the presence of other molecules (Damodaran 1997). As
can be seen from Fig. 3, the solubility of SCKPC was
clearly pH dependent. The maximum solubility
(92.96 ± 1.66%) was observed at pH 12.0 and it was
similar to the solubilities at pH 2.0 (85.52 ± 2.81%), 9.0
(86.26 ± 1.26%), 10.0 (90.15 ± 1.87%) and 11.0
(90.70 ± 2.20%) (p [ 0.05). Greater protein solubilities at
these pHs were likely associated with increased positive or
negative charges resulting in electrostatic repulsion (Moure
et al. 2001). The lowest solubility was at pH 5.0
(12.41 ± 1.23%), and the solubility at this pH was con-
siderably different from solubilities at other pHs
(p \ 0.05). As is the case in the present study, the majority
of food proteins are acidic and their minimum solubilities
are in the range of pH 4.0 to 5.0, and their maximum
solubilities are in alkaline pHs (Damodaran 1997). Shev-
kani et al. (2015) reported that cowpea protein isolates
showed the lowest protein solubility at pH 4.0–5.0, while
high protein solubility in acidic and alkaline pH.
The protein solubility of SCKPC in water at different
pHs showed a U-shaped curve (Fig. 3), similar to cashew
nut protein isolate (Ogunwolu et al. 2009), walnut protein
isolate and concentrate (Mao and Hua 2012), and cowpea
protein isolates (Shevkani et al. 2015).
Fig. 2 SDS-PAGE electrophoresis of sour cherry kernel products.
S Molecular weight standard, DSCKF defatted sour cherry kernel Water holding capacity
flour, SCKPC sour cherry kernel protein concentrate
The water holding capacity is regarded as an indicator of
a*(1.56 ± 0.47) and b* values, indicating the highest the ability of a protein to physically hold water against
lightness and the lowest redness and yellowness compared gravity (Kinsella 1981). WHC is a crucial functional
to the other samples. There was a significant increase property for highly viscous foodstuffs such as soups,
(p \ 0.05) in the L* value of DSCKF compared to SCKF, doughs, gravies, sauces, and bakery products. The water-
resulting probably due to the removal of lipid and lipid holding ability of the protein molecule is a function of size,
soluble pigments. On the other hand, the lowest L* value shape, hydrophilic and hydrophobic interactions. Further-
(p \ 0.05) was obtained for SCKPC, indicating that the more, water holding capacity is affected by lipid and car-
Maillard type browning reactions occurred probably during bohydrate content, and the properties of amino acid
drying at 50 °C. Similar result (a lower L* value) was residues on the surface (Damodaran 1997).
reported for chickpea protein concentrates (Ghribi et al. The WHC of SCKPC was 2.42 ± 0.09 g water/g
2015) because of longer drying time at 40 °C. L*, a* and (242%), being higher than those of pea protein isolate
b* values in the range of 62.9 to 77.6, 2.7 to 7.8 and 16.3 to (1.7 g/g), broad bean protein isolate (1.8 g/g), soy protein
19.4, respectively were reported for cowpea protein iso- isolate (1.3 g/g) (Fernandez-Quintela et al. 1997), bayberry
lates (Shevkani et al. 2015). kernel protein isolate (2.2 g/g) (Cheng et al. 2009), cashew
protein concentrate (1.74 g/g) (Ogunwolu et al. 2009), and
Protein solubility apricot kernel protein concentrate (1.40 g/g) (Sharma et al.
2010). However, it was lower than those of flaxseed protein
Protein solubility is a crucial factor for food applications concentrate (2.70 g/g) (Martinez-Flores et al. 2006), and
because it affects the other functional properties such as various rice bran protein concentrates (3.87–5.60 g/g)
emulsifying and foaming capacity, and serves as a useful (Chandi and Sogi 2007). Water holding capacity is a useful
indicator of denaturation and interactions of proteins. indication of whether protein concentrate or isolates can be

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Fig. 3 Solubility of sour cherry 100

kernel protein concentrate at
different pHs 90
80

Protein solubility (%)

70
60
50
40
30
20
10
0
1 2 3 4 5 6 7 8 9 10 11 12
pH

incorporated into aqueous food formulations. Higher WHC capacity may determine whether the protein concentrate or
of SCKPC suggests its appropriateness for improving the isolate can perform well as meat extenders or analogues. As
viscosity of food formulations. Hence, it could be useful in the SCKPC demonstrated satisfactory oil holding capacity,
extension of shelf life of meat products through the it could find application in sausages and cake batters.
reduction of moisture loss.
Foaming capacity and stability
Oil holding capacity
Foam formation and properties are influenced by the type
The interaction between protein and lipids determines the of protein, preparation method, composition, solubility,
sensory qualities of many foods. These interactions are concentration, pH, the presence of salts and hydrophobic
influenced by pH, ionic strength, temperature and the other interactions. Moreover, molecular elasticity, surface
variables in the system. Proteins with low solubility and hydrophobicity, charge distribution and hydrodynamic
high hydrophobicity can hold oil in large quantities properties affect foam formation and stability (Kinsella
(Damodaran 1997). High oil holding is essential for some 1981). Foaming capacity depends on the diffusion of sol-
food systems such as sausages, cake batters, mayonnaise uble proteins toward the air–water interface, rapid con-
and salad dressings (Chandi and Sogi 2007). formational change and rearrangement at the interface
The oil holding capacity of SCKPC was 1.73 ± 0.17 g (Damodaran 1997). Therefore, flexible protein molecules
oil/g (173%), being lower than that of sodium caseinate have a good foaming capacity. On the other hand, globular
(2.00 ± 0.04 g/g), but there were no significant difference proteins, whose surface denaturation is very difficult, have
(p [ 0.05). High OHC may indicate the presence of large low foaming capacity.
amounts of hydrophobic groups on the surface of the The foaming capacity of SCKPC at pH 7.0 was found to
protein molecule (Subagio 2006; Kaur and Singh 2007). be significantly lower than that of sodium caseinate used as
According to these results, there may be similar amount of a standard protein (p \ 0.05) (Table 2). This result indi-
hydrophobic groups on the surface of sour cherry proteins cated that the proteins in SCKPC was likely less flexible
and casein molecules. than sodium caseinate. The foaming capacity of SCKPC
The OHC of SCKPC was higher than that reported for soy (35.00 ± 3.54%) was lower than those of bayberry kernel
protein isolate (1.10 g/g) (Fernandez-Quintela et al. 1997), protein isolate (47.4%) (Cheng et al. 2009), cowpea protein
flaxseed protein concentrate (1.18 g/g) (Martinez-Flores isolates (82–93%) (Shevkani et al. 2015), and peanut pro-
et al. 2006), and apricot kernel protein concentrate (1.40 g/ tein concentrate (50%) (Wu et al. 2009). However, it was
g) (Sharma et al. 2010). However, this value was lower than higher than those of chickpea protein isolate (30.4%) (Kaur
that reported for hyacinth bean protein isolate (2.54 g/g) and Singh 2007), soy protein isolate (22.0%) (Fernandez-
(Subagio 2006), bayberry kernel protein isolate (1.80 g/g) Quintela et al. 1997), rice bran protein concentrate
(Cheng et al. 2009), peanut protein isolate (2.00 g/g) (Wu (5.2–8.7%) (Chandi and Sogi 2007), and apricot kernel
et al. 2009), cashew protein concentrate (3.32 g/g) (Ogun- protein concentrate (21.0%) (Sharma et al. 2010).
wolu et al. 2009), and chickpea protein concentrate Foaming stability is an important property since the
(1.91–2.77 g/g) (Ghribi et al. 2015). The oil holding usefulness of a foaming agent depends on its ability to

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Table 2 Foaming capacities

Properties (%) Sour cherry protein concentrate Sodium caseinate
and stabilities of sour cherry
kernel protein concentrate and Foaming capacity 35.00 ± 3.54a 87.50 ± 0.00b
sodium caseinate a
Foaming stability at 10 min 71.80 ± 7.25 83.62 ± 5.12a
a
Foaming stability at 30 min 71.80 ± 7.25 70.38 ± 17.82a
a
Foaming stability at 60 min 61.28 ± 11.24 50.71 ± 24.57a
a
Foaming stability at 90 min 54.10 ± 10.52 34.66 ± 8.16b
Foaming stability at 120 min 46.92 ± 9.79a 21.75 ± 0.67b
All the data are expressed as mean ± standard deviations and are the mean of three replicates (n)
a,b
Means followed by different letters within the same line represent significant differences (p \ 0.05)

maintain gas bubble for as long as possible (Kinsella 1981). probably did not adsorb to the water/lipid interface as
High foaming stability often requires a protein having rapidly as sodium caseinate. The EAI value of SCKPC
proper surface-active properties and adequate intermolec- (38.91 ± 2.50 m2/g) was much lower than those of hya-
ular interactions at the interface. The rise in the protein cinth bean protein isolate (534 m2/g) (Subagio 2006),
concentration of the continuous phase results in an increase chickpea protein concentrates (312.54–410.05 m2/g)
in the protein–protein interaction that leads to an increase (Ghribi et al. 2015) and flaxseed protein isolate (375 m2/g)
in viscosity. This facilitates the formation of an adhesive (Kaushik et al. 2016). However, it was higher than those of
multi-layer protein film around each gas bubble. The foams oat bran protein concentrates (with heat treatment
with this layer resist to liquid drainage and bubble coa- 18.90 m2/g, without heat treatment 20.04 m2/g) (Guan
lescence (Kaur and Singh 2007). et al. 2007), groundnut protein concentrate (5.43 m2/g)
Time-dependent (0–120 min) changes in the foaming (Jain et al. 2015), and cowpea protein isolates (7.7–8.9 m2/
stability of SCKPC are presented in Table 2. SCKPC had a g) (Shevkani et al. 2015).
lower stability than sodium caseinate after storage at room ESI is a measure of the ability of a protein to form a
temperature for 10 min, but had a higher stability after stable emulsion for a certain period (Subagio 2006). Sev-
30 min, although no statistically significant difference eral factors influence the emulsifying properties of proteins
existed between the values (p [ 0.05). The observed lower such as protein source, concentration, structural and sur-
foaming stability of SCKPC after storage at room tem- face characteristics, solubility, pH, temperature, and
perature for 10 min might be due to the fact that the ability equipment and methods used for forming emulsion. The
of SCKPC was limited to form a thick, cohesive and vis- emulsifying properties of protein concentrates are gener-
coelastic film around gas bubbles (Damodaran 1997). ally parallel to the water solubility profiles (Damodaran
Foaming stability of SCKPC (71.80 ± 7.25%) was higher 1997; Shevkani et al. 2015).
than those of cashew protein isolate (55%) (Ogunwolu The ESI value was found to be 1187.50 ± 17.70 min
et al. 2009) and bayberry kernel protein isolate (56%) for sodium caseinate, and 37.49 ± 2.41 min for SCKPC. It
(Cheng et al. 2009), and lower than those of soy protein was obvious that SCKPC had significantly lower ESI value
isolate (%93) (Fernandez-Quintela et al. 1997). than sodium caseinate (p \ 0.05). From this result, it was
inferred that the proteins in SCKPC had a limited capacity
Emulsifying activity and stability indices to reduce the interfacial tension and to form a protective
layer around the oil droplet. The ESI values of SCKPC
Surface hydrophobicity and concentration are the most (37.49 ± 2.41 min) was higher than that of peanut protein
important characteristics affecting the emulsifying ability concentrate (19.18 min) (Wu et al. 2009) and groundnut
of a protein. Also, a higher hydrophobic amino acid content protein isolate (28.62 min) (Jain et al. 2015). However, the
in the protein molecule favour emulsification (Damodaran values found for hyacinth bean protein isolate (2.7 h)
1997; Subagio 2006). (Subagio 2006) and flaxseed protein isolate (180 h)
EAI reflects the ability of a protein to adsorb rapidly to (Kaushik et al. 2016) were much higher than the result of
the water/lipid interface during emulsion formation. The the present study.
EAI values of SCKPC and sodium caseinate at pH 7.0 were
38.91 ± 2.50 m2/g and 176.44 ± 2.63 m2/g, respectively. The least gelling concentration
Sodium caseinate was used as a standard protein due to its
good emulsifying properties. As seen, the EAI value of The gelling properties of proteins are particularly important
SCKPC was significantly lower than that of sodium case- in emulsion meat products such as salami and sausage
inate (p \ 0.05), indicating that the proteins in SCKPC (Kinsella 1981). The gelling capacities of the proteins

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depend mainly on protein concentration, ionic strength, pH, the food industry as functional food ingredient. Further
and the amount of sulfhydryl and hydrophobic groups. In studies are required to determine the effect of various
addition, interactions of proteins with carbohydrates and protein extraction techniques on functional properties and
lipids also affect their ability to form a gel. Hydrogen yield of sour cherry kernel proteins.
bonds and ionic interactions are responsible for the stabi-
lization of the gel (Damodaran 1997). Physical, chemical Acknowledgements The study was partially supported by Tokat
Gaziosmanpaşa University Scientific Research Projects Unit (Project
or enzymatic applications can be used to form protein gels. No.: 2009/59).
The least gelling concentration is a measure of the gel
forming ability of a protein; the lower the LGC, the better
the gelling capacity. LGC of SCKPC at pH 7.0 was References
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Publisher’s Note Springer Nature remains neutral with regard to
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