FORMULA NUTRACEUTICAL PROTEIN COMBINATION OF FOUR TYPES OF

BEANS WITH HIGH DIGESTIBILITY AND GOOD FUNCTIONAL

CHARACTERISTICS

FORMULA NUTRASETIKAL PROTEIN KOMBINASI EMPAT JENIS KACANG

BERDAYA CERNA TINGGI DAN BERSIFAT FUNGSIONAL BAIK

Tejasari*, Sih Yuwanti, Nadia Ika Oktaviana

Faculty of Technology Agriculture, University of Jember
Jalan Kalimantan No.37 Tegalboto Campus, 68121, East Java, Indonesia
*Email: tejasari.unej@gmail.com

ABSTRACT

Digestibility can be determined by enzymatic biochemical analysis based on the nitrogen

content of amino acids in pea protein. Peanuts have high protein quality if they can be
hydrolyzed specifically or non-specifically to produce large amounts of amino acids. This study
aims to determine the quality and functional characteristics of the protein extract of peanuts and
the digestibility of peanut protein by combination treatment. This study used a completely
randomized design (CRD) with one factor, namely differences in the combination of peanut
nutraceutical protein. The test results showed that there was a significant effect on the quality
characteristics and functional properties of pea protein. The different combinations of peanut
nutraceutical protein used are considered to be able to improve the quality characteristics and
functional properties Analysis of protein digestibility using pepsin and pancreatin enzymes was
highest at 80 percent in the combined sample of mung beans and cowpea and the lowest was 65
percent in the lima beans, cowpea, and kidney beans combination samples. The protein content
of nutraceutical protein of beans was in the range of 69.4 – 94.0 percent. Functional
characteristics in the form of protein solubility of the pea nutraceutical protein were highest in
the sample combination of lima beans, cowpea, and kidney beans which was 99.7 percent and
the lowest were in the combined sample of mung beans and kidney beans, namely 50.8 percent.
The highest water absorption rate is 349.1 percent and the lowest is 210.6 percent. Oil absorption
ranged from 101.6 - 248.8 percent with the highest yield in the combined sample of lima beans
and mung beans and the lowest in the control sample of mung beans. The different combinations
of peanut nutraceutical protein used are believed to increase protein content, solubility, water
absorption, oil absorption, and protein digestibility.

Keywords: Digestibility, Protein, Peanuts

ABSTRAK

Daya cerna dapat ditentukan dengan analisis biokimiawi secara enzimatis berdasarkan
kandungan nitrogen asam amino dalam protein kacang. Kacang bermutu protein tinggi apabila
dapat dihidrolisis secara spesifik maupun non-spesifik untuk menghasilkan asam amino dalam
jumlah besar. Penelitian ini bertujuan untuk mengetahui karakteristik mutu dan fungsional
ekstrak protein kacang-kacangan serta daya cerna protein kacang dengan perlakuan kombinasi.
Penelitian ini menggunakan rancangan acak lengkap (RAL) dengan satu faktor yaitu perbedaan
kombinasi ekstrak protein kacang. Hasil pengujian menunjukkan terdapat pengaruh yang
signifikan pada karakteristik mutu dan sifat fungsional protein kacang. Perbedaan kombinasi
ekstrak protein kacang yang digunakan dianggap mampu meningkatkan karakteristik mutu dan
sifat fungsionalnya. Analisis daya cerna protein menggunakan enzim pepsin dan pankreatin
tertinggi sebesar 80 persen pada sampel kombinasi kacang hijau dan tunggak dan terendah
sebesar 65 persen pada sampel kombinasi kacang koro, tunggak, dan merah. Kadar protein
ekstrak kacang berada pada kisaran 69,4 – 94,0 persen. Karakteristik fungsional berupa kelarutan
protein ekstrak protein kacang tertinggi pada sampel kombinasi kacang koro, tunggak, dan merah
yaitu 99,7 persen dan terendah pada sampel kombinasi kacang hijau dan merah yaitu 50,8
persen. Tingkat penyerapan air tertinggi 349,1 persen dan terendah 210,6 persen. Daya serap
minyak berkisar antara 101,6 - 248,8 persen dengan hasil tertinggi pada sampel kombinasi
kacang koro dan tunggak dan terendah sampel kontrol kacang hijau. Perbedaan kombinasi
ekstrak protein kacang yang digunakan diyakini dapat meningkatkan kadar protein, kelarutan,
daya serap air, daya serap minyak dan daya cerna protein.

Kata kunci: Daya cerna, Protein, Kacang

INTRODUCTION

Digestibility needs set because determines the ability of a protein for broken down and
becomes amino acids or component of its constituents. Ability the determine protein quality.
Peanut high-quality protein could be hydrolyzed Specific or non-specific for producing amino
acids in a large amount. The number of amino acids in a protein determines nitrogen content.
Elemental nitrogen plays a role in the determination of digestibility so the number is very
influential in score digestibility. In general digestibility of vegetable protein is lower compared to
animal protein and has to limit amino acids so the diversification of protein sources expected
could increase protein quality. Quality protein height also has power and high protein
digestibility.
Digestibility determines the availability of amino acids by biological. Contains amino
acids in a protein determines big score percentage digestibility. If the protein can hydrolyze with
good Becomes amino acids so that the amounts of amino acids that are absorbed and utilized by
the body are high, then the digestibility of the protein high. Protein digestibility is the size
amount of Absorbed amino acids from a given protein intake. Measurement digestibility is based
on the measurement amount calculated nitrogen (N) residue as a protein that does not could
digest (Tejasari, 2019). Digestibility could be rated by biochemistry with the use of various
enzymes. Enzymes that can be used are pepsin-trypsin, pepsin-pancreatin, and solution
multienzyme (mixture of trypsin, chymotrypsin, and peptidase).
Potency protein amino acids in legumes are enough high. Contains amino acids in mung
beans including lysine 140.2 mg/g, methionine 130 mg/g, leucine 69.1 mg/g, and protein content
of 23.84% (Bishti et al., 2017). Peanut amino acids arrears include alanine 39 mg/g, arginine 75
mg/g, valine 50 mg/g (Rangel et al., 2004), and protein content of 20.5-22.11% (Research
Agency Peanuts and tubers, 2008). Lima bean's amino acids include serine 73.9 mg/g, histidine
32.4 mg/g, proline 76.2 mg/g (Chel-Guerrero et al., 2012), and protein content ranging from
between 19.93-21.40% (Diniyah et al., 2013). Kidney bean's amino acids include tryptophan
10.1 mg/g, phenylalanine 53.2 mg/g, and serine 4.72 mg/g (Audu and Aremu, 2011) with a
protein content of 23.10% (Nio, 2012). Content enough amino acids complete on fourth type
peanut this could utilize as product protein concentrate. Research previously has got digestibility
by enzymatic in peanut protein cowpea, lima beans, kidney beans, and mung beans (Tinus et al.,
2012; Siddhuraju, 2001; Nergiz, 2007; El-Adawy, 2000). The treatment combination of pea
protein expected capable increase score digestibility. In state mixed, amino acids derived from
pea protein different could each other fill and complete so that no there is no limiting amino acid.
Protein amino acids are also used for knowing the characteristics of protein function.
Characteristics of protein functional effect to the product to be generated. Because it is necessary
to conduct testing characteristics nature functional and value power protein digestibility
enzymatic with treatment combination of legume protein to use knowing influence combination
to score power protein digestibility and characteristics functional. Study this aim for knowing
protein content, characteristics of protein function (solubility, water absorption, and oil
absorption), and analysis of protein digestibility of legumes.

MATERIALS AND METHODS

Materials
Kidney beans, mung beans, lima beans, and cowpea are earned from the traditional
market Jember. After grinding each bean into 80 mesh fluor using a laboratory mill, the bean
flour was defatted with n-hexane for 1 h in a shaker. Enzyme pancreatin (Sigma-Aldrich
4xUSP), pepsin enzyme (Sigma-Aldrich 250 units/mg), aquadest, 1 M HCl (Mediss), NaOH
(Merck), trichloroacetic acid (TCA) (Merck), CuSO4.5H2O (Merck), sodium potassium tartrate,
Bovine Serum Albumin (BSA) (Sigma), Folin-Ciocalteu (Merck), Lowry mix, H 2SO4, selenium
(Merck), acid borate (H3BO3) (Merck), MM-MB (MBC) (Mediss), phosphate buffer pH 7
(Merck), another chemical namely n-hexane (Merck), palm oil, aluminum foil, and paper strain.
Specific tools used were a freeze dryer (CHRIST Alpha 1-2 LD plus), shaking water bath
(Stuart SBS 40), centrifuge (Tomy MRX-150 and Hitachi CR21GIII), pH meter (NAVI F-51),
spectrophotometer (Hitachi type U-2900 UV-Vis), Kjeldahl flask (BUTCHI); destilator
(BUTCHI K-355), and a burette.

Methods
Protein Extraction of the Bean
Protein extraction was performed using the isoelectric precipitation method (Chel-
Guerrero et al., 2012). The deffated flour suspension of beans (1:10 w/v for mung bean and
kidney bean) (1:6 w/v for cowpea dan lima bean) was adjusted to its optimal pH (pH 9 for mung
bean and kidney bean; pH 11 for cowpea and lima bean) by adding 1 M NaOH. After the pH
value is stable, do a stirring constantly for 1-2 hours at 25ºC using a magnetic stirrer. After that,
the solution was centrifuged (at 5500 rpm, 4ºC, 10 minutes for mung bean; 9500 rpm, 4ºC, 20
minutes for kidney; 10.000 rpm, 4ºC, 20 minutes for cowpea; and 10.000 rpm, 4ºC, 10 minutes
for lima bean) were done for obtaining the supernatant. The supernatant containing soluble
protein was gained thru subsequent precipitation by setting the isoelectric pH of each bean (pH
4,6 for mung bean and pH 4,6 for kidney, cowpea, and lima bean) with the addition of 1 M HCL
and stirring constantly until the pH is stable. Then let stand for 30 minutes to allow the protein to
be absorbed and perfectly deposited. The solution was centrifuged at the same condition. The
protein precipitate was washed twice with distilled water and then dried using a freeze-dryer for
the next analysis.

Protein Content Analysis of the Bean

 Protein content analysis is based on the Kjeldahl method which refers to Bakhtra et al.
(2017). The analysis is carried out by weighing samples as much as 0.1-0.5 g and then put into a
Kjeldahl flask. After that 5 ml of concentrated H 2SO4 and 0.9 g of selenium were added. The
solution is destroyed at a temperature of 410°C for approximately 2 hours or until clear and then
cooled. Prepared 15 ml of boric acid (H 3BO3) in an Erlenmeyer which was then added to 2 drops
of indicator mixture of methyl red and methyl blue. Then attached to a distillation apparatus, in
which the distillation tip must be immersed in boric acid. The sample of destruction that has been
made is then put in a flask distillation. The Kjeldahl flask was washed using sufficient aquadest
about 50 to 75 ml then attached to the distillation apparatus. The next step is NaOH 40% added
little by little from the top until the base (color slightly dark brown). The distillation process is
carried out until the distillate reaches approximately 40 ml. Yield solution distillation was then
titrated using 0.1 M HCl until the color changed. The percentage of protein was calculated as: %
N = (ml sample HCl-ml HCl blank) x M HCl x 14.01) / (sample weight x 1000) x 100% total
protein =% N x conversion factor (6.25). The sample and the blank were measured in their
protein content at three replications.

Protein Digestibility Analysis

Protein digestibility analysis was carried out enzymatically referring to Zhu et al. (2018)
and Kiin-Kabari et al. (2015) with a few modifications. The analysis begins by dissolving 150
mg of the sample with distilled water until the concentration of the solution is 3mg/ml and the
pH is adjusted to reach 1.5 using 0.1 N HCl. The solution was homogenized using a water bath
shaker at a speed of 50 rpm and a temperature of 37°C then added pepsin enzyme (5mg/ml in
0.01 M HCl) with a 1:100 ratio of enzyme and substrate was reacted for 30 minutes. After that, 1
M NaOH was added until the pH of the solution reached 7.8 then added the enzyme pancreatin
(5mg/ml phosphate buffer, pH 7) with a ratio of enzymes and substrates at 1:30. The reaction
was stopped after 60 min by adding 22.5 ml of 10% TCA solution. The solid residue is separated
by centrifugation for 5 minutes at 5000rpm, 5°C. The obtained solid was washed with 30 ml of
distilled water and filtered again with Whatman GF/C filter paper. The residue was filtered and
then dried in an oven at 105°C for two hours and analyzed for protein content (% residual
protein) using the Kjeldahl method. Protein digestibility was calculated as: % Digestibility =
(Amount of N in sample−Amount of N in residue) / Number of N in sample x 100%. The sample
was measured for its digestibility at three replications.

Protein Solubility Analysis

Protein solubility analysis was carried out referring to Zhu et al. (2017) with
modification. The bean protein nutraceutical was dissolved in distilled water (1% w/v) and the
pH was adjusted (to 3, 5, 7, 9, and 11) with 0.1 M HCl and 0.1 M NaOH while stirring for 30
minutes at 25ºC. The suspension was then centrifuged for 15 minutes at 5000 rpm. The total
protein in the hydrolyzates was analyzed by dissolving it in 0.2 M NaOH (1% w/v). The
supernatant obtained was then analyzed using a uv-vis spectrophotometer at 595nm. The content
of the dissolved protein in the supernatant and suspension as a whole was measured at three
replication using the Lowry method. The soluble protein was measured as percentage of the
soluble protein sample with soluble total protein.

Oil Absorption Analysis

 Oil absorption analysis was carried out referring to Mwangwela et al., (2007).
Measurements were made by weighing an empty centrifuge tube measuring 50 ml as the weight
of (a) grams, then 10 ml of palm oil is added and 0.5 g of sample was added and weighed as (b)
grams. Then the sample vortexed for 20 minutes at a speed of 2000 rpm until there was a
separation between supernatant and residue. The supernatant obtained was discarded and the
residue was weighed as weight (c) grams. Calculation of OHC can be calculated by the formula:
OHC (dB %) = ((weight C - weight A) − weight B) / weight B x 100%. Description A is the
weight of the empty cylinder, B is the weight of the cylinder containing, and C is the weight of
remaining precipitate. The sample measured its oil absorption at three replications.

Water Absorption Analysis

Analysis of water absorption was carried out referring to Mwangwela et al. (2007).
Measurements were made by weighing an empty centrifuge tube measuring 50 ml as weight (a)
grams, then 10 ml of distilled water was added, and added a sample of 0.5 g is weighed as (b)
grams. Then the sample is vortexed for 20 minutes at a speed of 2000 rpm until there is a
separation between supernatant and residue. The supernatant obtained was discarded and the
residue was weighed as weight (c) grams. WHC calculation can be calculated by the formula:
WHC (dB %) = (weight C - weight A) − weight B) / weight B x 100%. Description A is the
weight of the empty cylinder, B is the weight of the cylinder containing, and C is the weight of
remaining precipitate. The sample measured its oil absorption at three replications.

Statistical Analysis
Data from the test results of extract protein content, protein digestibility, solubility extract
protein, water absorption, and oil absorption were analyzed using SPSS software version 25.0
statistical method Analysis of Variance Test (ANOVA) with a significance level of = 0.05 or
95% confidence level. When it is significantly different, it will be continued with Duncan's
Multiple Range Test (DMRT) to determine the level of difference between treatments. Results
obtained are presented in tables and graphs. In addition, secondary data is also used obtained
from several references and several related studies.

RESULTS AND DISCUSSION

Protein Content of the Beans

Test protein levels performed for knowing protein content in nutraceutical Pea protein.
Protein content in extract peanuts is influenced by the nature physics, chemistry, and structure of
these proteins. Analysis of protein levels performed using method The calculated Kjeldahl based
on the total amount of nitrogen in the sample. The result of nutraceutical protein content obtained
from peanuts is presented in Figure 1
 Description:
K1: Mung Bean A2: MC Combination B1: MLC Combination
K2: Lima Bean A3: LC Combination B2: MCK Combination
K3: Cowpea A4: KC Combination B3: LCK Combination
K4: Kidney Bean A5: MK Combination B4: LKM Combination
A1: ML Combination A6: KL Combination C1: MLCK Combination
Figure 1. The protein content of nutraceutical protein of beans
Based on Figure 1 nutraceutical pea protein with treatment combination take effect on the
resulting protein content. The analysis of the protein content range between 69.4 ±0.99 – 94.0
±0.37 percent. Tiwari and Singh (2012) stated purity protein concentrate can reach 60-70 percent
and the composition is influenced by every method used. The protein content of each
nutraceutical peanut is similar, the treatment combination considered could increase the protein
content of peanuts.
Peanuts contain enough protein high. The study previously identify the nutraceutical
protein content mung beans at 72.0 percent, lima beans at 68.2 percent, cowpea at 64.2 percent,
and kidney beans at 62.5 percent (Afsari, 2018; Ahmad, 2018; Aulia, 2018; Agustina, 2018).
Based on literacy, protein content with treatment combination is considered to increase protein
content because could add nitrogen content in originating samples from peanut different.
Research obtained results of the highest protein content in the treatment combination of two
types of peanuts (mung beans and cowpea), this is due to control mung beans and cowpea taller
compared to lima beans and kidney beans so that treatment combination of two peanuts could
increase nutraceutical protein content.
The protein content is a basic parameter determination quality protein concentrate, high
protein content shows extraction performed effectively (Dennison, 2003). The protein content
produced by each nut is different depending on the amount of nitrogen it contains. Besides that
type of catalyst used and time heating, as well as addition ingredient reduction/oxidizing also
become influencing factors in extra protein content food (Muchtadi, 2010). Treatment
combination is considered capable increase protein content because the existence diversity
interlocking amino acids complete one each other so that Nitrogen levels also follow increase.
Protein Digestibility of Beans
The measure of the quality of a protein is not only determined by the protein content, but
also by the digestibility value. Protein digestibility shows the level of ease of protein to be
broken down into amino acids or their constituent components. The amino acids contained in a
protein determine big percentage score digestibility. Every type of peanut has power with
different digests. The analysis results in protein digestibility of pea and treatment the
combination by enzymatic (pepsin and pancreatin) can be seen in Figure 2

Description:
K1: Mung Bean A2: MC Combination B1: MLC Combination
K2: Lima Bean A3: LC Combination B2: MCK Combination
K3: Cowpea A4: KC Combination B3: LCK Combination
K4: Kidney Bean A5: MK Combination B4: LKM Combination
A1: ML Combination A6: KL Combination C1: MLCK Combination
Figure 2. Digestibility of nutraceutical protein of beans
Research results Based on Figure 2 show that the average digestibility obtained in the
whole sample is different by significance. Rated digestibility of nutraceutical protein of beans
highest A2 (combination of mung beans and cowpea) with a score of 80 ±0.35 percent
digestibility and lowest B3 (combination of lima beans, cowpea, and kidney beans) by 65 ±0.19
percent. This thing is following results which control shows that digestibility of mung beans and
cowpea is taller compared to kidney beans and lima beans. Based on the literature, amino acids
for example histidine in mung beans (37.5mg/g) and cowpea (37mg/g) are enough tall compared
to lima beans (32.4mg/g) and kidney beans (28.3mg/g) (Brishti et al., 2017; Rangel et al., 2004;
Chel-Guerrero et al., 2012; Audu and Aremu 2011).
Enhancement of digestibility is connected with an increase in total protein and a
decreased content of substance nutrients in the ingredients of food. Enhancement could be
caused by the hydrolysis process. The hydrolysis process used pepsin and pancreatin enzymes
capable break bond protein peptides Specific Becomes from amino acids so that digestibility the
taller (Shumoy et al., 2018). Protease enzymes break down proteins into free peptides and amino
acids so that could increase the concentration of amino acids. The enzyme pepsin converts
proteases to protease and peptone next followed by the hydrolysis process by enzymes capable of
pancreatin cut off chain peptide on side carboxyl protein (Muchtadi, 2010; Maurer, 2001).
 Based on Figure 2 treatment combination could increase the digestibility of pea protein
because in a state mixed derived amino acids of each bean could each other fill and complete
however, the amount is not far different with results digestibility one type peanuts. A study
previously obtained score digestibility by enzymatic in pea protein green by 80-85 percent (El-
Adawy, 2000), peanuts cowpea 82-86 percent (Tinus et al., 2012), beans red 68-72 percent
(Nergiz, 2007), and 68.25 percent lima beans (Sidduraju, 2001). The value of protein
digestibility can be affected by processing like heating and hydrolysis with sour so which could
reduce substance anti- nutrition in peanuts. Besides that, protein digestibility is influenced by
factors endogenous and exogenous. Factor exogenous is protein interactions with other
compounds such as polyphenols, carbohydrates, fats, and protease inhibitors. Whereas factor
endogenous is related to the characterization of protein structure (Caire-Juvera et al., 2013).

Solubility of the Bean Nutraceutical Protein

Protein solubility information is needed in protein-based food applications or processing.
Functional characteristics of protein in the form of solubility can affect the character of food and
are closely related to the type of protein. Proteins with high solubility can be widely applied
compared to proteins with low solubility. The results of protein solubility in nutraceutical protein
of various types of peanuts and their combinations can be seen in Figure 3
 Description:
K1: Mung Bean A2: MC Combination B1: MLC Combination
K2: Lima Bean A3: LC Combination B2: MCK Combination
K3: Cowpea A4: KC Combination B3: LCK Combination
K4: Kidney Bean A5: MK Combination B4: LKM Combination
A1: ML Combination A6: KL Combination C1: MLCK Combination
Figure 3. Solubility nutraceutical protein of beans
The results of the study based on Figure 3 showed that the combination of treatment and
pH conditions affected the solubility of the nutraceutical protein obtained. There was a
significant difference in the overall sample (p= 0.000 < 0.05). The lowest solubility value of
nutraceutical protein was found in sample A5 (combination of mung bean and kidney bean)
which was 50.8 ±0.21 percent, while the highest solubility value was found in sample B3
(combination of koro, cowpea, and kidney beans) which was 99.7 ±0.27 percent. Solubility of
nutraceutical protein of beans varies according to pH conditions and protein type. In Figure 3 can
be seen that the lowest pH conditions were found at pH 5. At that pH, the nutraceutical protein of
peanuts approaches point isoelectric pH 4-6 so that the protein does not have a charge and settle
(Li et al. 2005; Shevkani et al.,  2014; Khalid et al., 2013; Chel -Guero et al., 2012; Segura-
Campos et al., 2012).
On condition language more protein solubility tall compared to condition sour because
the amount of payload negative on the molecule more amino acids a lot. Based on Figure 3,
when condition acid and base sufficient protein solubility height, following the statement put
forward by Kusnandar (2010), protein reaches point solubility Lowest moment approach point
isoelectric, because at the point isoelectric interaction between protein molecules with other
proteins stronger compared protein interactions with water. Whereas when the pH is above or
under point isoelectric protein interacts with more water strong so that protein can dissolve
better.
In general, protein solubility is influenced by several factors, including pH, ionic
strength, and molecular size. Differences in the solubility level of nutraceutical protein in beans
can also be caused by differences in the number and length of amino acid chains as well as
differences in the ratio of hydrophilic amino acids to hydrophobic amino acids (Moure et al.,
2006). Increasing the pH of the extraction is thought to be able to extract polar amino acids, the
further the pH of the solution is from the isoelectric pH, the solubility of amino acids will
increase. According to research by Kempka et al., (2014) protein has a very wide utilization
application related to its functional and nutritional properties by considering solubility as a
specific property for use in food processing, however, protein extracts that show high solubility
results in acidic pH need to be limited in use. Protein solubility affects the processing of food
products, the higher the solubility, the better for food products.

Water absorption nutraceutical protein of beans

Water absorption is useful for measuring the ability of food material to retain the water it
absorbs. Protein can bind water because it is hydrophilic. The results of the analysis of the water
absorption of the protein extract of beans can be seen in Figure 4
 Description:
K1: Mung Bean A2: MC Combination B1: MLC Combination
K2: Lima Bean A3: LC Combination B2: MCK Combination
K3: Cowpea A4: KC Combination B3: LCK Combination
K4: Kidney Bean A5: MK Combination B4: LKM Combination
A1: ML Combination A6: KL Combination C1: MLCK Combination
Figure 4. Water absorption nutraceutical protein of beans
The results based on Figure 4 show that the average water absorption obtained in all
samples is significantly different (p= 0.000 < 0.05). Figure 4.4 shows that the water absorption of
the extract ranges from 210.6 ±0.06 - 349.1 ±0.13 percent. There were significant differences in
the three samples treated with the combination of two beans (mung bean and cowpea), (cowpea
and lima beans), a combination (kidney beans and lima beans), and a combination of three beans
(green, koro, and kidney beans). The highest absorption was found in sample A3 (lima beans and
cowpea) and the lowest was in sample K1 (mung bean). The combination of nutraceutical
peanuts can increase the amount of protein so that the value of water absorption increases. The
increase in absorption value can also be caused by the different types of amino acids contained in
each peanut sample. Onsaard (2012) stated that amino acid composition, protein conformation,
and protein surface polarity affect water absorption.
Proteins are hydrophilic due to the presence of peptide chains that have polar groups such
as carbonyl, hydroxyl, amino, carboxyl, and sulfhydryl, so they can form bonds with water. The
number and type of different polar groups cause the protein's ability to absorb water to be
different (Triyono, 2010). In Figure 4 the value of water absorption in the combination of lima
beans and cowpea is high due to the presence of charged amino acids such as lysine, aspartic
acid, and glutamic acid in large quantities. This is following the literacy of Rosida et al. (2015),
which state that the important amino acids found in cowpea are the amino acids lysine, aspartic
acid, and glutamic acid. According to literacy, the number of amino acids in lima beans and
cowpea, respectively, is; lysine (59.9 mg/g and 68 mg/g), aspartic acid (128 mg/g and 116 mg/g),
glutamic acid (153 mg/g and 162 mg/g) (Rangel et al. 2004; Chel-Guerrero et al. 2012).
Hydrophilic amino acids in legumes include lysine, arginine, serine, threonine, tyrosine,
histidine, aspartic acid, and glutamic acid (Brishti et al. 2017; Rangel et al. 2004; Chel-
Guerrero et al. 2012; Audu and Aremu 2011). The content of amino acids which is quite large in
the peanut extract should be able to increase the value of water absorption in the combination
treatment of three and four types of beans. The results of the analysis obtained in this study are
not following the literature proposed by Prabowo (2010), that the higher the protein, the greater
the water absorption, and the lower the protein, the lower the water absorption.

Oil absorption nutraceutical protein of beans

Functional properties of proteins whose mode of action tie oil-free that is oil absorption.
Oil absorption is used to measure the ability of a protein in withhold absorbed oil. Oil absorption
takes effect on the natural texture and quality of food. Analysis result power absorb nutraceutical
protein oil and treatment the combination could be seen in Figure 5

Description:
K1: Mung Bean A2: MC Combination B1: MLC Combination
K2: Lima Bean A3: LC Combination B2: MCK Combination
K3: Cowpea A4: KC Combination B3: LCK Combination
K4: Kidney Bean A5: MK Combination B4: LKM Combination
A1: ML Combination A6: KL Combination C1: MLCK Combination
Figure 5. Oil absorption nutraceutical protein of beans
Based on Figure 5, can is known that treatment combinations have a significant influence
on the absorption of the resulting oil (sig = 0.000 < 0.05). Figure 5 shows that oil absorption in
nutraceutical protein peanuts range between 101.6 ±0.06 - 248.8 ±1.67 percent. Rated power
absorb oil highest found in sample A3 (combination of lima beans and cowpea) and the lowest
in-sample K2 (lima beans). Rated oil absorption is different in each sample could cause because
to the existing differences in protein content, amino acids, and non-polar components of protein.
This thing in line with the statement of Yoshie et al. (2008) that oil absorption is related to the
composition and type of amino acids. Oil absorption could influence by several factors including
protein content, non-polar protein components, and levels of liquidity from oil. Oil absorption
can also be caused by the size of a particle, the size of more particles small and uniform will
produce high oil absorption content (Suwarno, 2003).
In Figure 5 it is known oil absorption treatment control is worth lower compared with
treatment combination extract peanuts. This is following the statement of Sutrisniati (1995) in
Giyarto et al. (2016) that a higher protein content will result in more oil absorption. The value of
oil absorption is closely related to hydrophobicity and protein solubility. Proteins that have a
high absorption value are hydrophobic, hydrophobic proteins are effective at lower surface
tension and bind more lipophilic materials.
The increase in the value of oil absorption can be caused by the combination treatment,
but it is also due to the different types of amino acids contained in each peanut sample. The
increase in protein content also increases oil absorption due to its hydrophobicity. Similar to
water absorption, oil absorption depends on the protein structure of the ingredients. The protein
structure which is more lipophilic (non-polar) contributes to the increase in oil absorption (Lin et
al., 1974 Pratiwi et al., 2018). Onsaard (2012) also suggested that the mechanism of oil
absorption is caused by the presence of non-polar groups on proteins that are bound to the
hydrocarbon chains of fatty acids or oils. The hydrophobic amino acids in legumes include
alanine, glycine, leucine, valine, isoleucine, phenylalanine, proline, cysteine, methionine, and
tryptophan (Brishti et al., 2017; Rangel et al., 2004; Chel-Guerrero et al., 2004). al., 2012; Audu
and Aremu 2011).

CONCLUSION

The protein content of legume extracts was in the range of 69.4 – 94 percent with the
highest value in sample K1 (mung beans) and the lowest in-sample K2 (lima beans). Functional
characteristics of protein extracts from legumes had the highest solubility value in sample B3
(combination of koro, cowpea, and kidney beans) of 99.7 percent and the lowest in sample A5
(combination of green and kidney beans) of 50.8 percent. The water absorption value of the
protein extract of legumes ranged from 210.6 – 349.1 percent with the highest value in sample
A3 (combination of lima beans and cowpeas) and the lowest in-sample K1 (mung bean).
Functional characteristics in the form of oil absorption of the protein extract of peanuts obtained
the highest value of 248.8 percent in sample A3 (combination of lima beans and cowpeas) and
the lowest value of 101.6 percent in sample K2 (lima beans). The digestibility of the protein
extract of legumes ranged from 65 – 80 percent with the highest value in sample A2
(combination of green and cowpeas) and the lowest in sample B3 (combination of koro, cowpea,
and kidney beans).

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