583 Egypt. J. Chem. Vol. 66, No. 12, pp.

353 - 365 (2023)

Egyptian Journal of Chemistry

http://ejchem.journals.ekb.eg/

Angiotensin-converting enzyme (ACE-1) inhibitory and

antioxidant activities of probiotic yogurt enriched with rice bran

Mohamed Samir Darwish *1, Mohamed Shalaby Gomaa 1, Elsherbiny S.

Elsherbiny 1 and Mohamed S. Mostafa 1
Dairy Department, Faculty of Agriculture, Mansoura University, Mansoura 35516, Egypt

Abstract

Angiotensin converting enzyme inhibitory (ACE-I) peptides make up the majority of the bioactive peptides created
during yogurt production. In the current study, the influences of rice bran (RB) fortification at two different levels
(1 and 2%) on several quality aspects of probiotic set yogurt were assessed during 14 days of cold storage. Probiotic
yogurts were analyzed and contrasted with plain yogurt in terms of their in vitro ACE-I inhibitory activity,
proteolysis, and antioxidant capability, as well as various physicochemical and sensory evaluations. The TS, TSS,
syneresis, apparent viscosity, and water holding capacity values of yoghurt samples were positively impacted by
RB addition up to 2% concentration. All yogurt batches had higher antioxidant, O-phthalaldehyde and ACE-
inhibitory activity up until day 7, but they started to decline at day 14. The maximum activity was seen in yogurt
with 2% RB during 7 days of storage, while control yogurt had the lowest antioxidant and ACE-inhibitory activity
at zero time (p ˂ 0.05). The panelists gave all RB-fortified yogurts lower ratings than they gave plain yoghurt. The
viability of S. thermophilus, B. lactis BB-12 and L. acidophilus LA-5 was improved by the addition of various
concentrations of RB. To produce dairy products with increased antioxidant and improved anti-ACE properties,
rice bran may be employed to change the microbial fermentation of milk.

Keywords ACE- inhibitory activity, total phenolic content, antioxidant activity, proteolysis and rice bran

1. Introduction in the previous studies of probiotic yogurt [1]. The

The utilization of probiotic organisms [Lactobacillus least 106 CFU/mL of probiotic bacteria is the
acidophilus and Bifidobacteria] in food, especially concentration of live cells that is most frequently
dairy products, has garnered considerable interest in advised for probiotic products [2,3]. Relying on the
recent years. One of the fermented milk products that peptides created through fermentation and
is widely consumed globally is yogurt, which contains cold storage, probiotic yogurt may have enhanced
probiotic bacteria, along with other dairy products like health benefits. Given that the prevalence of
yogurt drinks. Due to its beneficial features for health, hypertension is rising rapidly, recently, peptides that
such as preventing allergies, immune system could really reduce blood pressure among
activation, treating diarrhea, restoring intestinal hypertensive people have garnered a lot of attention
microbiata and lowering cholesterol, one popular [4]. These peptides' mechanism of action is based on
functional food is probiotic yoghurt. Additionally, the suppression of the ACE-1, but it cannot be ruled
different probiotic bacteria strains that are utilized to out that their activity may also involve a variety of
make yogurt may help to produce bioactive peptides other complex pathways that could have additional
that have anticancer, antihypertensive and antioxidant positive impacts on consumer health [5]. The enzyme
properties [1]. ABT cultures which have Lactobacillus ACE is especially necessary for the rennin-angiotensin
acidophilus, Bifidobacteria and Streptococcus system, which adjusts the balance of salt and water and
thermophilus as main fermenting organisms are blood pressure in the body [5]. Angiotensin I is
generally used in the production of probiotic yogurt. hydrolyzed by ACE into a potent vasoconstrictor
As well as this, L. acidophilus, Lactobacillus casei and (angiotensin II), which results in an increase in blood
Lactobacillus paracasei have been studied extensively pressure and degradation to a larger extent than
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*Corresponding author e-mail: [email protected].; (Mohamed Samir Darwish).
Receive Date: 28 January 2023, Revise Date: 18 March 2023, Accept Date: 08 April 2023, First Publish Date: 09 April 2023
DOI: 10.21608/EJCHEM.2023.190160.7527
©2023 National Information and Documentation Center (NIDOC)
354 Mohamed Samir Darwish et.al.
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necessary of bradykinin, which has vasodilatory matter content, crude protein content, crude fat
activity [6]. content, mineral content and crude fiber content [22].
Prebiotics were not digestible food ingredients and In a nutshell, method 962.09 was used to calculate the
have positive effects on host health due to their crude fiber content, the ash content was calculated
characteristics that are responsible for activation or using method 942.05, and the moisture content was
reproduction of probiotic bacteria in colon [7,8]. estimated using method 934.01. The protein content
Numerous studies have been conducted to determine was calculated using the Kjeldahl method, which was
the effects of enriching milk with prebiotic substances computed as nitrogen x 6.25 (method 2001.11). The
to encourage the probiotics growth. The best way to amount of crude fat was assessed by Soxhlet
fortify milk with prebiotics is to add fiber-rich foods extraction. utilizing a flame atomic absorption
like nuts, grains and fruits to it [3,9-12]. Foods spectrophotometer Agilent 240 FS (Agilent
containing both prebiotics and probiotics are referred Technologies, USA), levels of minerals were
to as synbiotic foods [13]. In addition to synbiotic is measured after the instrument was calibrated with the
the synergy between prebiotic and probiotic [14]. The appropriate standard solutions in accordance with the
most common way that probiotic bacteria are applied official procedure established by the Association of
is through dairy products, and yoghurt has been Official Analytical Chemists.
successfully marketed and modified to appeal to the
target consumers. For a modulated production of 2.3. Polysaccharides extraction:
yogurt, several way may be applied such as prebiotics The rice bran powdered was pre-extracted by Soxhlet
addition or probiotic culture selection [15]. There are system for 48 h with acetone, followed by extraction
several researches were concerned about the synbiotic with methanol for 48h, the supernatant was eliminated.
fermented milk [16,17]. The precipitate was dried at 50 °C for 72h. the residue
The outer layer of rice, known as rice bran (RB), is a was added to hot distilled water (50-60 °C) at 1:10
byproduct of the milling process [18]. Depending on g/ml for 1h and subsequently was centrifugated for 20
the source and milling methods, rice bran's chemical min at 10. 000, supernatant was kept, and three further
make-up differs. It contains 25 to 45% carbohydrate, extractions were performed on the precipitate to
12 to 25% oil 10 to 16% protein, 6 to 10% ash 6 to extract any remaining polysaccharides. The mixed
15% crude fiber [19]. Traditionally, it has been used supernatants were centrifugated for 20 min at 12.000g,
mostly in animal feeds, but in recent years, its use in the pellet was removes and the combined supernatants
human food and nutrition has increased dramatically were added to ethanol (final concentration 80% (v/v))
[20]. More focus has been placed on rice bran and the precipitate was dried at 50 °C for 24 h and
polysaccharides (RBPSs) among these beneficial RB dissolved in appropriate distilled water [23].
components because of their many biological
advantages, such as their anti-inflammatory, 2.4. Investigation of rice bran's monosaccharide
antioxidant and anti-tumor properties, as well as the content:
enhancement of immune function and diarrhea The analysis of monosaccharide composition of crude
symptoms [20,21]. Therefore, the current study aimed polysaccharide of rice bran was determined with slight
to examine the effects of combining rice bran with modification according to [24]. Briefly, 1 g rice bran
milk on yogurt with respect to (1) yogurt acidification, polysaccharide were added to 100 ml trifluoro acetic
(2) sensory and physicochemical properties, (3) acid (4 M), followed by heated at 80 °C for 12 h and
antioxidant activity, (4) milk protein hydrolysis and subsequently filtered by syringe filter (0.45 µm).
(5) ACE-1 inhibition. HPLC (Agilent, USA) supplied with quaternary pump,
refractive index (RI detector) operated at 40 °C and
2. Materials and Methods: column (300 mm x 7.8 mm operated at 80 °C) used
2.1. Materials: phenomenex ® Rezex RCM-Monosaccharide. The
Raw cow milk with 12% total solids, 3.0% fat, 3.0% separation process is carried out using isocratic elution
protein, and a pH of 6.70, as well as rice bran, were by flow rate 0.6 ml/min with HPLC grade water.
bought at local markets in Mansoura City, Egypt. The
ABT-2 culture consisted of Bifidobacteria lactis BB- 2.5. Functional yogurt production:
12, Streptococcus thermophilus ST-20 and L. To produce functional yogurt, fresh raw cow milk was
acidophilus LA-5 and was obtained by Chr. Hansen, used. First, milk was separated into three experimental
Egypt. lots, each of which contained 1 and 2% RB powder
with the exception of plain yogurt. Each mixture was
2.2. Analyses of the rice bran's chemical heated for 15 minutes at 94 °C before being quickly
composition and minerals: cooled to 40 ± 1 °C in chilled water for the 0.03%
The chemical composition analysis of rice bran was (w/w) addition rate of the freeze-dried ABT-2 culture.
determined using the methods of the Association of All experimental yogurts were divided among 100 mL
Official Analytical Chemists, comprising the dry sterile plastic containers, which were then sealed and
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Egypt. J. Chem. 66, No. 12 (2023)
 Angiotensin-converting enzyme (ACE-1) inhibitory and antioxidant activities of probiotic yogurt enriched with rice bran 355
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kept at 4 °C after being incubated at 40 ± 1°C until a 2.9. Sensory evaluation of yogurt samples:
pH of 4.4–4.5 was obtained. A previously described technique by Darwish, Qiu,
Taher, Zaki, Abou-Zeid, Dawood, Shalabi, Khojah
2.6. The procedure for making yogurt water and Elawady [9] was slightly modified to quantify the
extracts: sensory evaluation of yoghurt samples. In a nutshell,
2.5 mL of sterile deionized water were mixed with 10 all yoghurt samples underwent a consumer approval
g of yogurt samples. The yoghurts' pH was measured, test (hedonic sensory assessment) after being kept cold
and then they were acidified with HCl to pH 4.0. (0.1 for an overnight period, seven days, and fifteen days.
M). The acidified mixtures were then centrifuged The majority of the untrained panelists were Mansoura
(4500g, 12 min, 5 °C) after being heated in a water University staff members and students, all of whom
bath for 15 min at 45 °C. NaOH (0.1 M) was added to consumed yoghurt on a regular basis. The ratings were
the supernatant to bring its pH up to 7.0. The given on a 9-point hedonic scale, with extreme like on
neutralized supernatants were centrifuged again for the right receiving a score of 9, neither like nor dislike
12 minutes at 5 °C (4500 g), and until it was required on the center receiving a score of 5, and severe dislike
for analysis, the supernatant was gathered and kept at on the left receiving a score of 1. The consumer
20 °C in a freezer [9]. panelists evaluated the body and texture, sourness,
appearance, odor, taste and overall acceptance of the
2.7. Determination of total titratable acid (TTA) products. To avoid bias, the panelists were not given
and pH: any information regarding the different sample types.
A titration with 0.1 N NaOH was used to calculate
TTA. 9 mL of dH2O and 1 mL of sample were 2.10. O-Phthalaldehyde (OPA) method:
blended. As a pH indicator, A few drops of The peptide content of yogurt water extract was
phenolphthalein at 0.1% were applied. The mixture examined using the spectrophotometric OPA-based
was then continually stirred while being titrated with technique [33]. Prepared ingredients included 0.95 g
0.1 N NaOH until a steady pink color emerged. The of sodium tetraborate, 25 mL of dH2O, and 20 g/100
following formula was used to determine how much mL (wt/wt) sodium dodecyl sulphate. 100 mL of -
acid was created during fermentation [25]. Before mercaptoethanol were added after dissolving 40 mg of
determining pH, Initially, water was used to OPA in 1 mL of methanol. In a volumetric flask, 25
homogenize the yoghurt (1: 9 ratio). Using a digital pH mL of sodium tetraborate solution, 20 g/100 mL
meter (Hanna Hi2210), The homogenized sample's pH (wt/wt) SDS, and OPA mixture were combined. The
was measured. [26]. volume was then brought up to 50 mL by adding sterile
Lactic acid percentage = V NaOH x 0.009 x 0.1 N x dH2O. 30 mL of yogurt water extract were put into a
dilution factor (10) x 100…. (1) plastic cuvette along with 1 mL of OPA reagent. After
two minutes, a reading at 340 nm was recorded. A340
2.8. Physicochemical analysis of yogurt samples: was converted to a standard curve based on tryptone.
The method of AOAC [27] was used to measure the
total solid and titratable acidity of yogurts, and a 2.11. ACE-inhibitory activity analysis:
refractometer was used to determine the total soluble According to Elbermawi, Darwish, El-Awady, Zaki,
solids content. Yogurt samples' pH levels were Qiu and Samra [10], with a small adjustment, the
measured using the technique of Metin [28]. Using a inhibitory activity was evaluated
chroma meter (Konica Minolta, Inc., Osaka, Japan), spectrophotometrically. The process relies on ACE
the a (redness), b (yellowness), and L (lightness; 0 = cleaving hipuryl-histdylleucine (HHL) to hippuric
black and 100 = white) values of yoghurt samples were acid. HHL (3.8 mM) was diluted in 200 µL of 0.1 M
determined [29]. Yogurt syneresis was found in borate buffer, which contains 0.3 M NaCl and has a
accordance with Farooq and Haque [30]. A test tube pH of 8.3, and then 35 µL of the yogurt water extract
containing about 10 g of yogurt was centrifuged at was added. The combination was then incubated for
5000 rpm for 20 min at 4 C to determine the water 10 min at 37 °C. 20µ L of ACE solution (0.1 U/mL in
holding capacity (WHC), which was then expressed as borate buffer) was added to start the reaction, which
(clear supernatant/initial weight) X 10 [31]. he was then left to run for 30 min at 37 °C. The reaction
apparent viscosity of the samples was measured using was then put into a 250 µL solution of 1 M HCl to put
a viscometer (Brookfield Rotational Viscometer). an end to it. Hippuric acid was obtained by mixing 1.5
Peroxide value (PV) analysis was performed using the mL of ethyl acetate vigorously for 30 seconds and then
acetic acid-chloroform method (AOCS Cd 8-53) and centrifuging the mixture at 1200 g for 10 minutes. The
the findings were represented as milliequivalent solvent was then removed by placing 1 mL of the
peroxide/kg product [32]. supernatant (ethyl acetate) in boiling water for 30
minutes in a fresh tube. To test absorbance at 228 nm,
the residual hippuric acid residue was dissolved in 1
mL of distilled water. Distilled water served as a
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Egypt. J. Chem. 66, No. 12 (2023)
356 Mohamed Samir Darwish et.al.
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control. Each sample was examined three times. fiber. Potassium was the most prevalent mineral in the
Lyophilized aqueous extract (whey) produced from rice bran sample's mineral makeup (2.03 ± 0.11%),
plain yogurt was utilized as a negative control whereas followed by phosphorus and magnesium (1.23 ± 0.1
captopril (0.008 mg/mL) was used as a positive and 0.67 ± 0.09%, respectively), while minerals
control. The formula for calculating ACE inhibitory detected in the minor percentages were calcium,
activity was as follows: manganese, iron, zinc, sodium and copper (390.33 ±
ACE inhibition (%) = 1-[(A-C)/(B-D)] X 100…… 2.1, 158.733 ± 2.4, 71.43 ± 0.76, 50.87 ± 1.6, 19.73 ±
(2) 1.74 and 2.42 ± 0.55 mg/Kg, respectively)
wherein A represents absorbance with the sample,
ACE and HHL; B represents absorbance with ACE Table (1). The proximate composition and mineral
and HHL without sample; C represents absorbance content of rice bran
with the sample and HHL; and D represents Parameter Value
absorbance with HHL without ACE and sample. Proximate composition (%)
Dry matter 91.15 ± 0.61
2.12. Enumeration of viable bacteria: Ash 8.23 ± 0.72
In yogurt samples taken at various points during Crude fiber 6.36 ± 0.79
storage, viable bacterial counts were counted. Crude protein 12.74 ± 0.87
Different selective media were employed to count the Crude fat 16.15 ± 0.85
microorganisms. By pour plating 1 mL of each Mineral content
dilution in M-17 agar (Oxoid) and incubating under K (g/100g) 2.03 ± 0.11
aerobic circumstances at 39 ± 1 °C for 48 hours, the P (g/100g) 1.23 ± 0.09
viability of S. thermophilus was evaluated [34]. MRS- Mg (g/100g) 0.67 ± 0.08
NNLP agar, which includes paramycin sulphate (200 Ca (mg/Kg) 390.3 ± 2.1
mg/L), lithium chloride (3000 mg/L), neomycin Fe (mg/Kg) 71.43 ± 0.76
sulphate (100 mg/L), and nalidixic acid (150 mg/L), Mn (mg/Kg) 158.73 ± 2.39
was used to count B. lactis BB-12. Just before pouring, Zn (mg/Kg) 50.87 ± 1.6
filter sterilized NNLP (NNLP; Sigma Chemical Co.) Na (mg/Kg) 19.73 ± 1.74
was added to the autoclaved MRS foundation. To Cu (mg/Kg) 2.42 ± 0.55
reduce the medium's oxidation-reduction potential and Results are mean of three values ± Standard error (SD)
promote the development of anaerobic bifidobacteria,
0.05% final concentration of filter-sterilized L- 3.2. Composition of monosaccharide:
cysteine-HCl was also added at the same time to the Monosaccharide compositions of RBP was assessed
medium. Triplicate inoculated plates were by HPLC. The results indicated that glucose was
anaerobically incubated at 37°C for 72 hours [35]. L. presented as the essential monosaccharide of RBP and
acidophilus LA-5 viable counts were counted on BL- the percentage of glucose, galactose, arabinose,
MRS (agar enriched with 1.5 g/l bile salts) [36]. They mannose and xylose was 34.44, 10.11, 3.15, 2.69 and
underwent anaerobic incubation for 72 hours at 37 C. 4.12% (Table 2). These results was contestant with
The counts of viable bacteria expressed as log other reports of Liu, Cao, Zhuang, Han, Guo, Xiong
CFU/mL. and Zhang [20], who showed that monosaccharides of
RBP are composed of glucose, galactose, xylose,
2.13. Statistical analysis: mannose and arabinose at a molar ratio of 32:6:5:2:4.
Every test was run in triplicate. The total titratable acid This result also partly agreed with Zha, et al. [37], who
(TTA), pH, physicochemical parameters, sensory found out three types of polysaccharides in hot
properties, OPA, TPC, antioxidant, and ACE- aqueous extracts from rice bran and determine their
1inhibitory activities were all evaluated using an compositions of monosaccharides according to GC
ANOVA test with a significance level of p ˂ 0.05. The and GC-MS analysis, PW1 was consisted of glucose,
information was displayed as an average and standard galactose, mannose, arabinose and ribose with molar
deviation. The Duncan's multiple range tests were percentage of 54.1%, 21.7%, 10.5%, 6.3% and 7.4%
used to identify significant deviations between values. respectively, while the molar percentage of PW3 was
All statistical tests in the current study were evaluated 50.7%, 32.7%, 10.1%, 4.1% and 2.4%. Wang, et al.
using SPSS Statistics program. [38] also detected that the polysaccharide of rice bran
was consisted of xylose, arabinose, galactose and
3. Results and discussion: glucose at molar ration of 2:4:4:1. The variation
3.1. Proximate and mineral composition of rice among the compositions of monosaccharide of RBP
bran may be due to contradiction between raw materials,
The chemical composition of rice bran is shown in steps of purification and protocols of extraction.
Table (1). Rice bran contains 16.15 ± 0.85% fat, 12.74
± 0.87% protein, 8.23 ± 0.72% ash and 6.63 ± 0.80%
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 Angiotensin-converting enzyme (ACE-1) inhibitory and antioxidant activities of probiotic yogurt enriched with rice bran 357
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Table (2): Monosaccharides composition of rice bran

polysaccharides
Parameter Value
Glucose 34.44 ± 0.70
Galactose 10.11 ± 0.55
Arabinose 3.15± 0.35
Mannose 2.70 ± 0.30
Xylose 4.12 ± 0.61
Results are mean of three values ± Standard error (SD)

3.3. Effects on rice brane on yogurt fermentation:

Initial pH measurements for all yogurts were similar Fig. (1): pH levels of yogurts during the process of
(6.59-6.61; Fig. 1). During the 90–180-minute fermentation in the presence or absence of 1% and
fermentation period, yogurts fortified with RB 2% of rice bran (RB) at 41 °C. Vertical bars represent
experienced pH lowering rates that were faster (0.47 ± the means of treatments ± standard deviations.
0.05 and 0.52± 0.03 pH unit/h for 1 and 2% RB,
respectively) than control (0.34 ± 0.05 pH unit/h). As
a consequence, yogurt fortified with 2% RB was the
first to achieve pH 4.5 (298 min), followed by 1% RB
and plain yogurts (327 and 400 min, respectively).
Rice bran appeared to increase the yogurt bacteria's
metabolic activity. The pH scale measures the amount
of H+ that is present as a result of lactic acid bacteria
(LAB) producing organic acids. By using a typical
starting culture, yogurt was fermented thanks to the
symbiotic interactions of two bacterial groups.
According to Sandine and Elliker [39], Formic acid Fig. (2): Alterations in total titratable acid (% lactic
and carbon dioxide, among other S. thermophilus acid) during yogurt fermentation at 41 C in the
metabolites, encourage the growth of certain presence or absence of 1% and 2% rice bran (RB).
Lactobacillus spp., whereas amino acids and short Vertical bars represent the means of treatments ±
peptides produced by Lactobacillus spp. proteolytic standard deviations.
activity promote the growth of S. thermophilus [40].
Yogurts' pH dropped to lower pH levels (4.2 - 4.4) 3.4. Physicochemical properties of yogurt samples:
after being refrigerated for 28 days, potentially After 7 days, samples' physicochemical characteristics
because of the accumulation of acetic acid, were evaluated, and the findings are displayed in Table
acetaldehyde, formic acid, and lactic acid [41]. TTA is (3). Yogurt containing 2% RB was shown to have the
displayed in Fig. (2) and represents the proportion (%) highest levels of total soluble solids (TSS) and total
of lactic acid that is found in yogurt during solids (TS), while plain yogurt had the lowest levels of
fermentation. Compared to pH changes represented by these parameters (Table 3). According to Demirci, et
the “S” saped- curve, the increase in TTA for all yogurt al. [46], adding more rice bran to formulation of
samples was practically linear. At each stage of the yogurt dramatically enhanced TS and TSS levels.
incubation, the TTA for the yogurt samples fortified A key flaw in yogurt products, syneresis, is caused by
with various concentrations of RB was higher than that an extreme lack of curd stability. As shown in Table
of plain yogurt. Between the 90th and 210th minute of (3), the rice bran-based yogurts showed lower
fermentation, yogurt fortified with 2% of RB produced syneresis levels than plain yogurt. This could be
TTA at the greatest rate (0.213 ± 0.007), followed by explained by the fact that RB-fortified yogurt may
yogurt with 1% of RB and plain (0.193± 0.004 and have improved water-holding properties due to the
0.1567 ± 0.003%/h, respectively). The total amount of dietary fibers found in RB, including pectin, ß-glucan,
hydrogen ions in the sample of yogurt, excluding those arabinogalactan, hemicellulose, and
bonded to alkaline ions, is known as total titratable galactooligosaccharide. Additionally, Hu and Yu [47]
acidity. Thus, when evaluating the starter culture's demonstrated that dietary fibers from RB, including
capability for fermentation, TTA determination is hemicellulose and insoluble fibers, have a range of
more crucial [42]. According to Østlie, et al. [43], the beneficial properties, such as a high swelling capacity
production of lactic, formic, butyric, acetic, butyric, and capacity for holding water, which allowed them to
and citric acid in yogurt is linearly associated with bind more water and have a harder texture.
the TTA accumulation [44]. Accordingly, differences Furthermore, oat and barley ß-glucan significantly
in microbial population during fermentation can be reduced separation of whey in yogurts containing B.
linked to changes in yogurt's titratable acidity [45]. bifidum, according to Ozcan and Kurtuldu [48].
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358 Mohamed Samir Darwish et.al.
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Table (3) contains the samples' viscosity values. The the rising b values (Table 3). The similar finding
set yogurt appeared to have less apparent viscosity regarding when wheat bran was added to yoghurt, the
after addition of RB. All of the RB added yogurts had L* value fell or the a* and b* values rose [46].
viscosities that were lower than plain yogurt. This is
most likely a result of stabilizing casein aggregates and 3.5. Sensory evaluation of yogurt samples:
covering caseins with polysaccharides like pectin and The results that adding RB to yogurt samples had a
ß-glucan. These findings concur with those of Tseng negative impact on the sensory aspects in terms of
and Zhao [32], who found that adding wine grape overall preference, taste, sourness, body texture, odor
pomace as a prebiotic to yogurt decreased its perceived and appearance (Fig. 3). The panelists gave all RB-
viscosity values. Additionally, El-Said, et al. [49] fortified yogurts lower ratings than they gave plain
reported that decreasing viscosity values were yogurt. Increased rice bran content in this study may
achieved by increasing the concentration of have decreased yogurt acceptance. In agreement with
pomegranate peel extracts. They linked these findings the present study, Demirci, Aktaş, Sözeri, Öztürk and
to how the extract affected the electrostatic Akın [46] found that yogurts with RB scored lower on
aggregation of networks in yogurts. However, yogurts the sensory scale than plain yogurt. Yogurts with more
with inulin and peach dietary fiber added showed fiber had better body and texture, but their overall
considerably greater apparent viscosities than plain flavor quality decreased, according to Fernández-
yogurt [50,51]. Garía, et al. [53]. Similar to this, Hashim, et al. [54]
The amount of additional rice bran caused an increase found that yogurt that has been enriched with date
in peroxide levels (PV). At day 7, The PV for plain fiber at different concentrations and 1.5% wheat bran
yogurt was the lowest (1.06 mequiv/kg), while the PV did not receive higher overall sensory scores or be
for 2% RB yogurt was the highest (1.96 mequiv/kg) more palatable. As seen in Fig. (3), the yogurt fortified
(Table 3). This outcome was in line with observation with 1% RB demonstrated increased acceptance when
Sanabria [52] that adding purple rice bran oil boosted compared to the other fortified yogurt. Lack of a ritual
the PV value of frozen yogurt. of adding cereal-based additions to yoghurt among the
The presence of RB had an impact on the L* values (p volunteers, even though most consumers are aware of
˂ 0.05). RB-containing yogurt samples had lower L* the health advantages of these materials, may be a
values than the control group. L* values rapidly fell as contributing factor to the comparatively low
RB addition increased (Table 3). Additionally, RB preference scores reported for RB enrichment.
added samples had greater a* and b* values than plain
yogurt. The yellowish tint of RB can be used to explain

Table (3): Physicochemical characteristics of yogurt samples

Parameters Plain yogurt 1% RB 2% RB
Total solid (%) 12.64 ± 0.18 c 13.51 ± 0.17 b 14.10 ± 0.16 a
Total soluble solids (%) 10.08 ± 0.20 c 10.63 ± 0.09 b 11.20 ± 0.23 a
c
WHC (%) 38.35 ± 0.43 41.22 ± 0.31 b 43.68 ± 0.51 a
a
Syneresis (% v/w) 10.63 ± 0.09 9.75 ± 0.12 b 8.28 ± 0.27 c
a
Apparent viscosity (cP) 3177 ± 87.8 1966 ± 152.8 b 1620 ± 203 b
c
Peroxide (mequiv./Kg) 1.05 ± 0.03 1.27 ± 0.05 b 1.74± 0.07 a
* a
L 89.23 ± 0.11 86.14 ± 0.29 b 83.33 ± 0.38 c
* c
b 6.26 ± 0.12 10.20 ± 0.22 b 11.79 ± 0.32 a
* c
a -2.90 ± 0.15 -1.14 ± 0.10 b -0.46 ± 0.12 a
Significant differences are denoted by lowercase letters in the same row (p ˂ 0.05).

Fig. (3): Sensory scores of yogurt samples fortified with different concentration of RB compared with plain
yogurt after storage period for 7 days at 5±2°C. Lowercase letters present a significant impact on the sensory
assessment. Vertical bars represent the means of treatments ± standard deviations.
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 Angiotensin-converting enzyme (ACE-1) inhibitory and antioxidant activities of probiotic yogurt enriched with rice bran 359
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3.6. Total phenolic content: specific phytochemical contents of rice bran and the
Yogurt fortified with 2% of RB had the highest (TPC) results of microbial metabolic activities most likely
content (30.23 ± 0.87 mgGEA/mL), followed by contributed to the increased antioxidant activities in
yogurt fortified with 1% of RB and plain yogurts (20.4 yogurt samples supplemented with RB than in plain
± 0.85 and 11.17 ± 0.8 mgGEA/mL, respectively) yogurt [59]. Even at low temperatures, yoghurt
(Fig. 4). During refrigerated storage, the TPC bacteria are metabolically active, may be responsible
considerably (p ˂ 0.05) rose for all yogurt samples, for the maximum inhibition of DPPH inhibition
reaching maximum values of 38.4 ± 0.87, 28.17 ± following seven days of storage. Several of the
0.96, and 16.93 ± 0.70 mgGEA/mL in yogurts that antioxidant effects of polyphenols may have changed
were 2% RB, 1% RB, and plain yogurt, respectively because of ongoing microbial development during
(Fig. 4). Due to the absence of any plant extracts in chilled storage. Several of the antioxidant effects of
plain yogurt, the TPC values in control represent polyphenols may have changed as a result of ongoing
phenolic substances linked to the breakdown of milk microbial development during chilled storage [57]. It
protein [55]. Tyrosine, an amino acid, is one example is believed that increased breakdown of phenolic
where a phenolic side chain has been proposed to components with antioxidant properties is what caused
contribute to the reading in TPC [56]. Another idea is the decline in antioxidant activity during yogurt
that during fermentation and post-acidification, preservation in the refrigerator [60] or increasing the
bacteria use phenolic acids like p-coumaric acid and interaction between polyphenol and milk protein [61].
ferulic to produce other phenolic acids like p- To take advantage of the strong antioxidant activities
hydroxybenzoic acids and vanillic before the aromatic beneficial for a preventive cardiovascular effect,
ring structure is broken down [57]. The presence of yogurt consuming is highly advised within 7 days
native phytochemical substances (such as flavonoids following yogurt production [62].
and phenolic compounds) in rice bran can be used to
explain the higher TPC in yogurt supplemented with
RB [58].

Fig. (5): Response surface plot for antioxidant ability

Fig. (4): TPC in yogurt supplemented with RB when of yogurt fortified with different concentration (1 and
stored in a refrigerator (4 °C) compared with control. 2%) of rice bran and plain yogurt over 14 days of
Values are means ± SD after three replications of refrigeration (5°C) storage (% suppression of DPPH
each experiment. Gallic acid equivalent (ugGAE)/mL oxidation). Values are means ± SD from three
units were used to express the total phenols. separate experiments. Significant changes in
Significant changes in TPC between storage period antioxidant activity between storage period
conditions are indicated by lowercase letters (p ˂ conditions are indicated by lowercase letters (p ˂
0.05). Significant TPC changes across treatment are 0.05). Significant changes in antioxidant activity
denoted by capital letters (p ˂ 0.05). across treatment are denoted by uppercase letters (p ˂
0.05).
3.7. DPPH scavenging activity of yogurt samples:
Following fermentation and when being stored, the 3.8. Proteolysis evaluation by o-phthalaldehyde
antioxidant activity of all yogurt samples fortified with (OPA) method:
RB was higher (p ˂0.05) than that of plain yogurt (Fig. On the initial storage day, all yogurt samples that had
5). On day 7 of cold storage, each yogurt's ability to been fortified with RB had OPA values that were
inhibit DPPH oxidation climbed to its greatest levels, higher (17.3–18.5 mg/mL) than those of control (15.33
which were 60.87 ± 0.91, 51.2 ± 1.4, and 28.27 ± mg/mL) (Fig. 6). Seven days after being stored, yogurt
1.22% for RB-2, Rb-1 and plain yogurt, respectively. that had been fortified with RB at various
By day 14, the amount of inhibition had gradually concentrations showed an increase in proteolysis
decreased between 6.5 and 15.5% (Fig. 5). The (52.6–96%), with the highest peptide concentrations

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Egypt. J. Chem. 66, No. 12 (2023)
360 Mohamed Samir Darwish et.al.
_____________________________________________________________________________________________________________

being seen at 2% of RB (36.2 ± 1.2 mg/mL) followed of bioactive peptides created during the production of
by 1% of RB (26.4 ± 1.02 mg/mL). The OPA levels in yogurt and refrigeration storage. The level of ACE
yogurt fortified with RB (23.63 ± 1.0 and 18.9 ± inhibition peaked on day 7 of storage but subsequently
0.62mg/mL for 2 and 1% of RB, respectively) were decreased between 5.26 and 15.16 % on day 14 of
higher than those in plain yogurt (15.6 ± 1.06 mg/mL) storage (Fig. 7), corresponded with comparable
on day 14 of storage. Since S. thermophilus and L. reductions in OPA values. This indicates that during
acidophilus are metabolically active even at 4 °C [5], the first seven days of cold storage, the relatively less
the presence of different concentrations of rice bran particular peptides created throughout fermentation
may have increased the amount of proteolysis by these have been further cleaved to smaller, more bioactive
bacteria, as evidenced by a significant rise in OPA proteins, and that extensive proteolysis of these
values in yogurt after the first seven days of chilled proteins during prolonged storage up to day 14
storage (Fig. 6). This might change protein breakdown produced much smaller, less bioactive proteins.
and possibly increase the generation of bioactive Amirdivani and Baba [25] found that produce
peptides [63]. Over the following 14 days, the OPA fermented dairy products with stronger anti-ACE
value in plain yogurt during refrigeration remained properties, the microbial fermentation of milk may be
largely stable, whereas it decreased in yogurt fortified modified using peppermint, dill, and basil. At all
with different concentration of RB. Proteolysis was storage times, yogurt samples fortified with RB at
first accelerated by both RB concentrations before varying concentrations displayed greater anti-ACE
being slowed down. One potential explanation for this activity than control (p ˂ 0.05). The maximum ACE
is because the RB promotes cell metabolism growth, inhibition was seen in yogurt fortified with 2% of RB,
which raises the acidity [64]. As a result, the lactic acid followed by yogurt with 1% of RB. By attaching to
generated had an impact on the yogurt proteins, ACE, biomolecules like phytochemicals can change
causing denaturation and aiding hydrolysis. However, the enzyme's activity [25]. According to Amirdivani
the hydrolysis resulted in the production of peptides and Baba [25], since the ACE activities were
and free amino acids, which reduced the acidity. unaffected by the water extract of peppermint, dill, and
Similar to this, once the bacteria started consuming the basil, these herbs may have indirectly changed the
generated organic acids and peptides after using up the degradation of milk proteins by influencing the
sugar resources, proteolysis decreased [65]. metabolism and growth of the starter culture in
yogurt.

Fig. (6): O-phthalaldehyde (OPA) levels in plain and

yogurt fortified with RB changed following
fermentation (41 °C) and throughout 28 days of
chilled storage (4 °C). Values are means ± SD from
three separate experiments. Significant changes in
OPA between storage time conditions are indicated
by lowercase letters (p ˂ 0.05). Significant variations Fig. (7): Inhibition of the angiotensin-I converting
in OPA between treatment doses are denoted by enzyme (ACE-I) by a water extract of RB fortified
capital letters (p ˂ 0.05). yogurt stored at 4°C compared with control. Values
are means ± SD from three separate experiments.
3.9. ACE-inhibitory activity: Significant changes in ACE-1 between storage period
By attaching biomolecules like polyphenols, conditions are indicated by lowercase letters (p ˂
flavonoids, and bioactive peptides to the enzyme's 0.05). Significant variations in ACE-1 between
active binding sites, biomolecules like ACE's treatment concentrations are denoted by uppercase
enzymatic activity can be changed [66]. The ACE letters (p ˂ 0.05).
assay was used to assess the ACE-inhibitory activity
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Egypt. J. Chem. 66, No. 12 (2023)
 Angiotensin-converting enzyme (ACE-1) inhibitory and antioxidant activities of probiotic yogurt enriched with rice bran 361
__________________________________________________________________________________________________________________

3.10. Counts of viable microorganisms: WHC, whereas a decrease syneresis of set yogurt. RB
The changes in the B. lactis BB-12, S. thermophilus supplementation at different concentrations enhanced
ST-20 and L. acidophilus LA-5 counts obtained in of total phenolic content and antioxidant activity of
yogurt samples during storage are shown in Fig.8. Up yoghurt samples. Unfortunately, acceptance of all RB-
to 14 days of storage, probiotic counts for L. enriched yoghurts was lower than control. The
acidophilus LA-5 and B. lactis BB-12 were typically probiotic yogurt's inclusion of rice bran had no
greater than the minimal therapeutic threshold (106 negative influence on the bacterial counts. In contrast,
CFU/ g). In all yogurts with or without RB, S. it improved the probiotic strains' ability to survive. (L.
thermophilus levels ranged from 6.1to 9.28 log CFU/g. acidophilus LA-5 and B. lactis BB-12) during cold
S. thermophilus counts in samples of RB-added storage. These RB fortified yogurts at different
yoghurt gradually declined between days 1 and 14 of concentration, by virtue of the increased antioxidant
storage. Our findings are consistent with those of activities and existence of bioactive peptides and may
Espírito Santo, et al. [67], who discovered a provide a novel range of yogurts with eligible
comparable decline in S. thermophilus counts in açai multifunctional health benefits to consumers with
yoghurts co-fermented with B. lactis B104 between 1 hypertension.
and 14 days. The findings agreed with those of
Saccaro, et al. [68] and Senaka Ranadheera, et al. [69],
who noted a small decline in S. thermophilus during
the storage of probiotic yoghurts with fruit juice and L.
acidophilus LA5.
Throughout storage, the presence of RB with different
concentrations had significant effect (p ˂ 0.05) on the
number of B. lactis BB-12 counts (Fig. 8). The yogurt
with 2% RB and yogurt with plain yogurt had the
lowest and greatest concentrations of B. lactis BB-12,
respectively. At the end of the shelf life, the final
counts of B. lactis BB-12 were 6.73 ± 0.06 and 7.27 ±
0.12, respectively. The count of B. lactis BB-12 in Fig. (8): Microbial changes of RB fortified yogurt
yoghurt fortified with 1% and 2% RB rose gradually and control during 14 days of cold storage. Vertical
up to 7 days of cold storage period. With regard to L. bars represent the means of treatments ± standard
acidophilus LA5 counts, after d1 both 1 and 2 % RB deviations. Significant changes in viable counts of
fortified yogurt samples showed counts significantly bacteria between storage period conditions are
higher than plain yogurt (p ˂ 0.05). All microbial indicated by uppercase letters (p ˂ 0.05). Significant
counts were negatively impacted by the longest shelf variations in microbial counts between treatments are
life (d 14), with B. lactis BB-12 showing by far the denoted by lowercase letters (p ˂ 0.05).
lowest values. A synergistic effect between different
concentration of RB (1-2%) and probiotic strains L. Data Availability Statement
acidophilus LA5 and B. lactis BB-12) is evident (p < All of the data is contained within the article and the
0.05). Similar to this, researchers who examined at Supplementary Materials.
various yogurt supplements noticed that L. acidophilus
or Bifidobacteria levels rose [70,71]. Santos, et al. [72] Conflicts of Interest
had also studied the prebiotic flours used in dairy food The authors declare no conflict of interest.
processing and emphasized the possible prebiotic
benefits of adding flour derived from fruits or other 5. References
sources by encouraging the growth and increasing the 1. Erkaya-Kotan, T. In vitro angiotensin converting
viability of probiotics like Lactobacillus and enzyme (ACE)-inhibitory and antioxidant
Bifidobacteria. activity of probiotic yogurt incorporated with
orange fibre during storage. Journal of Food
4. Conclusion Science and Technology 2020, 57, 2343-2353,
The presence of rice bran modified the yogurt bacteria doi:10.1007/s13197-020-04272-1.
fermentation of milk leading to improved yogurt 2. Lourens-Hattingh, A.; Viljoen, B.C. Yogurt as
acidification, bioactive peptides formation and ACE probiotic carrier food. International Dairy
inhibition. In the presence of 2% RB, followed by 1% Journal 2001, 11, 1-17,
RB, yogurt bacteria had the highest levels of doi:https://doi.org/10.1016/S0958-
proteolytic activity during fermentation and cold 6946(01)00036-X.
storage. Higher peptide synthesis during storage is 3. Darwish, M.S.; Abou-Zeid, N.A.; Khojah, E.; Al
well correlated with increased ACE inhibition. Jumayi, H.A.; Alshehry, G.A.; Algarni, E.H.;
Fortification of RB resulted in an increase in pH, Elawady, A.A. Supplementation of Labneh with
________________________________________________
Egypt. J. Chem. 66, No. 12 (2023)
362 Mohamed Samir Darwish et.al.
_____________________________________________________________________________________________________________

Passion Fruit Peel Enhanced Survival of E. coli Be Used as Powerful Tool to Improve Cell
Nissle 1917 during Simulated Gastrointestinal Surfaces Properties and Adherence Potential of
Digestion and Adhesion to Caco-2 Cells. Foods Probiotic Strains. Fermentation 2022, 8, 122,
2022, 11, doi:10.3390/foods11111663. doi:10.3390/fermentation8030122.
4. FitzGerald, R.J.; Murray, B.A.; Walsh, D.J. 13. Pandey, S.M.; Mishra, H. Optimization of the
Hypotensive Peptides from Milk Proteins. The prebiotic & probiotic concentration and
Journal of Nutrition 2004, 134, 980S-988S, incubation temperature for the preparation of
doi:10.1093/jn/134.4.980S. synbiotic soy yoghurt using response surface
5. Papadimitriou, C.G.; Vafopoulou- methodology. LWT-Food Science and
Mastrojiannaki, A.; Silva, S.V.; Gomes, A.-M.; Technology 2015, 62, 458-467.
Malcata, F.X.; Alichanidis, E. Identification of 14. Zubaidah, E.; Nurcholis, M.; Wulan, S.N.;
peptides in traditional and probiotic sheep milk Kusuma, A. Comparative Study on Synbiotic
yoghurt with angiotensin I-converting enzyme Effect of Fermented Rice Bran by Probiotic
(ACE)-inhibitory activity. Food Chemistry 2007, Lactic Acid Bacteria Lactobacillus casei and
105, 647-656, Newly Isolated Lactobacillus plantarum B2 in
doi:https://doi.org/10.1016/j.foodchem.2007.04. Wistar Rats. APCBEE Procedia 2012, 2, 170-
028. 177, doi:10.1016/j.apcbee.2012.06.031.
6. Coates, D. The angiotensin converting enzyme 15. Soccol, C.R.; Pandey, A.; Larroche, C.
(ACE). The International Journal of Fermentation processes engineering in the food
Biochemistry & Cell Biology 2003, 35, 769-773, industry; CRC Press: 2013.
doi:https://doi.org/10.1016/S1357- 16. Granata, M.; Brandi, G.; Borsari, A.; Gasbarri,
2725(02)00309-6. R.; Gioia, D.D. Synbiotic yogurt consumption by
7. Mattila-Sandholm, T.; Myllärinen, P.; healthy adults and the elderly: the fate of
Crittenden, R.; Mogensen, G.; Fondén, R.; bifidobacteria and LGG probiotic strain.
Saarela, M. Technological challenges for future International journal of food sciences and
probiotic foods. International Dairy Journal nutrition 2013, 64, 162-168,
2002, 12, 173-182. doi:10.3109/09637486.2012.718742.
8. Oliveira, R.P.; Florence, A.C.; Silva, R.C.; 17. Hussein, L.; Gouda, M.; Fouad, M.; Labib, E.;
Perego, P.; Converti, A.; Gioielli, L.A.; Oliveira, Bassyouni, R.; Mohammad, M. Dietary
M.N. Effect of different prebiotics on the intervention with yoghurt, synbiotic yogurt or
fermentation kinetics, probiotic survival and traditional fermented sobya: bio-potency among
fatty acids profiles in nonfat symbiotic fermented male adolescents using five bio-markers of
milk. International Journal of Food relevance to colonic metabolic activities. Food
Microbiology 2009, 128, 467-472. and Nutrition Sciences 2014, 2014,
9. Darwish, M.S.; Qiu, L.; Taher, M.A.; Zaki, A.A.; doi:10.4236/fns.2014.512123.
Abou-Zeid, N.A.; Dawood, D.H.; Shalabi, 18. Ryan, E.P. Bioactive food components and
O.M.A.K.; Khojah, E.; Elawady, A.A. Health health properties of rice bran. Journal of the
benefits of postbiotics produced by E. coli Nissle American Veterinary Medical Association 2011,
1917 in functional yogurt enriched with Cape 238, 593-600.
gooseberry (Physalis peruviana L.). 19. Sharma, H.R.; Chauhan, G.S.; Agrawal, K.
Fermentation 2022, 8, 128-150, Physico-Chemical Characteristics of Rice Bran
doi:10.3390/fermentation8030128. Processed by Dry Heating and Extrusion
10. Elbermawi, A.; Darwish, M.S.; El-Awady, A.A.; Cooking. International Journal of Food
Zaki, A.A.; Qiu, L.; Samra, R.M. Isolation and Properties 2004, 7, 603-614, doi:10.1081/JFP-
biological activities of compounds from Rumex 200033047.
vesicarius L. and their use as a component of a 20. Liu, Q.; Cao, X.; Zhuang, X.; Han, W.; Guo, W.;
synbiotic preparation. Food Chemistry: X 2022, Xiong, J.; Zhang, X. Rice bran polysaccharides
14, 100306, doi:10.1016/j.fochx.2022.100306. and oligosaccharides modified by Grifola
11. Elbermawi, A.; Darwish, M.S.; Zaki, A.A.; frondosa fermentation: Antioxidant activities and
Abou-Zeid, N.A.; Taher, M.A.; Khojah, E.; effects on the production of NO. Food Chemistry
Bokhari, S.A.; Soliman, A.F. In Vitro 2017, 223, 49-53,
Antidiabetic, Antioxidant, and Prebiotic doi:https://doi.org/10.1016/j.foodchem.2016.12.
Activities of the Chemical Compounds Isolated 018.
from Guizotia abyssinica. Antioxidants 2022, 11, 21. Choi, J.-y.; Paik, D.-J.; Kwon, D.Y.; Park, Y.
doi:10.3390/antiox11122482. Dietary supplementation with rice bran
12. Khojah, E.; Gomaa, M.S.; Elsherbiny, E.G.; fermented with Lentinus edodesincreases
Elawady, A.; Darwish, M.S. The In Vitro interferon-γ activity without causing adverse
Analysis of Postbiotics in Functional Labneh to effects: a randomized, double-blind, placebo-
________________________________________________
Egypt. J. Chem. 66, No. 12 (2023)
 Angiotensin-converting enzyme (ACE-1) inhibitory and antioxidant activities of probiotic yogurt enriched with rice bran 363
__________________________________________________________________________________________________________________

controlled, parallel-group study. Nutrition Dairy Technology 2003, 56, 26-29,

Journal 2014, 13, 35, doi:10.1186/1475-2891- doi:https://doi.org/10.1046/j.1471-
13-35. 0307.2003.00070.x.
22. Horwitz, W.; Latimer Jr, G. Association of 32. Tseng, A.; Zhao, Y. Wine grape pomace as
Official Analytical Chemists–AOAC. Official antioxidant dietary fibre for enhancing
methods of analysis of AOAC International. 18th nutritional value and improving storability of
ed. Gaithersburg: AOAC International 2006. yogurt and salad dressing. Food Chemistry 2013,
23. Hefnawy, H.T.M.; El-Shourbagy, G.A. 138, 356-365,
Chemical analysis and antioxidant activity of doi:https://doi.org/10.1016/j.foodchem.2012.09.
polysaccharide extracted from rice bran. World 148.
Journal of Dairy & Food Sciences 2014, 9, 95- 33. Shori, A.B.; Baba, A.S. Effects of inclusion of
104. Allium sativum and Cinnamomum verum in milk
24. Dawood, D.H.; Elmongy, M.S.; Negm, A.; on the growth and activity of lactic acid bacteria
Taher, M.A. Extraction and chemical during yogurt fermentation. American-Eurasian
characterization of novel water-soluble Journal of Agricultural & Environmental
polysaccharides from two palm species and their Sciences 2013, 13, 1448-1457.
antioxidant and antitumor activities. Egyptian 34. Sah, B.N.P.; Vasiljevic, T.; McKechnie, S.;
Journal of Basic and Applied Sciences 2020, 7, Donkor, O.N. Effect of probiotics on antioxidant
141-158, and antimutagenic activities of crude peptide
doi:https://doi.org/10.1080/2314808X.2020.177 extract from yogurt. Food Chemistry 2014, 156,
3126. 264-270,
25. Amirdivani, S.; Baba, A.S. Changes in yogurt doi:https://doi.org/10.1016/j.foodchem.2014.01.
fermentation characteristics, and antioxidant 105.
potential and in vitro inhibition of angiotensin-1 35. Tharmaraj, N.; Shah, N.P. Selective Enumeration
converting enzyme upon the inclusion of of Lactobacillus delbrueckii ssp. bulgaricus,
peppermint, dill and basil. LWT - Food Science Streptococcus thermophilus, Lactobacillus
and Technology 2011, 44, 1458-1464, acidophilus, Bifidobacteria, Lactobacillus casei,
doi:https://doi.org/10.1016/j.lwt.2011.01.019. Lactobacillus rhamnosus, and Propionibacteria.
26. Taher, M.A.; Khojah, E.; Darwish, M.S.; Journal of Dairy Science 2003, 86, 2288-2296,
Elsherbiny, E.A.; Elawady, A.A.; Dawood, D.H. doi:https://doi.org/10.3168/jds.S0022-
Biosynthesis of Silver Nanoparticles by 0302(03)73821-1.
Polysaccharide of <i>Leucaena 36. Lima, K.G.d.C.; Kruger, M.F.; Behrens, J.;
leucocephala</i> Seeds and Their Anticancer, Destro, M.T.; Landgraf, M.; Gombossy de Melo
Antifungal Properties and as Preservative of Franco, B.D. Evaluation of culture media for
Composite Milk Sample. Journal of enumeration of Lactobacillus acidophilus,
Nanomaterials 2022, 2022, 7490221, Lactobacillus casei and Bifidobacterium
doi:10.1155/2022/7490221. animalis in the presence of Lactobacillus
27. AOAC, H.W. International A: Official Methods delbrueckii subsp bulgaricus and Streptococcus
of Analysis of the AOAC International. The thermophilus. LWT - Food Science and
Association: Arlington County, VA, USA 2000. Technology 2009, 42, 491-495,
28. Metin, M. Analysis Methods of Milk and Dairy doi:https://doi.org/10.1016/j.lwt.2008.08.011.
products (Sensory, Physical and Chemical 37. Zha, X.-Q.; Wang, J.-H.; Yang, X.-F.; Liang, H.;
Analysis). Bornova: Ege University Publications Zhao, L.-L.; Bao, S.-H.; Luo, J.-P.; Xu, Y.-Y.;
2006. Zhou, B.-B. Antioxidant properties of
29. Pinho, O.; Mendes, E.; Alves, M.M.; Ferreira, polysaccharide fractions with different molecular
I.M.P.L.V.O. Chemical, Physical, and Sensorial mass extracted with hot-water from rice bran.
Characteristics of “Terrincho” Ewe Cheese: Carbohydrate Polymers 2009, 78, 570-575.
Changes During Ripening and Intravarietal 38. Wang, L.; Zhang, H.; Zhang, X.; Chen, Z.
Comparison. Journal of Dairy Science 2004, 87, Purification and identification of a novel
249-257, doi:https://doi.org/10.3168/jds.S0022- heteropolysaccharide RBPS2a with anti-
0302(04)73163-X. complementary activity from defatted rice bran.
30. Farooq, K.; Haque, Z.U. Effect of sugar esters on Food chemistry 2008, 110, 150-155.
the textural properties of nonfat low calorie 39. Sandine, W.E.; Elliker, P.R. Microbially induced
yogurt. Journal of Dairy Science 1992, 75, 2676- flavors and fermented foods. Flavor in fermented
2680. dairy products. Journal of Agricultural and Food
31. Celik, S.; Bakirci, I. Some properties of yoghurt Chemistry 1970, 18, 557-562.
produced by adding mulberry pekmez 40. Kailasapathy, K. Survival of free and
(concentrated juice). International Journal of encapsulated probiotic bacteria and their effect
________________________________________________
Egypt. J. Chem. 66, No. 12 (2023)
364 Mohamed Samir Darwish et.al.
_____________________________________________________________________________________________________________

on the sensory properties of yoghurt. LWT - Food 52. Sanabria, L.A.A. Development of a frozen yogurt
Science and Technology 2006, 39, 1221-1227, fortified with a nano-emulsion containing purple
doi:https://doi.org/10.1016/j.lwt.2005.07.013. rice bran oil; Louisiana State University and
41. Vedamuthu, E.R. Fermented Milks. Di dalam Agricultural & Mechanical College: 2012.
Rose, AH Economic Microbiology. 1982. 53. Fernández-Garía, E.; McGregor, J.U.; Traylor, S.
42. Geidam, Y.A.; Ambali, A.G.; Onyeyili, P.A. The Addition of Oat Fiber and Natural
Phytochemical screening and antibacterial Alternative Sweeteners in the Manufacture of
properties of organic solvent fractions of Psidium Plain Yogurt1. Journal of Dairy Science 1998,
guajava aqueous leaf extracts. International 81, 655-663,
Journal of Pharmacology 2007, 3. doi:https://doi.org/10.3168/jds.S0022-
43. Østlie, H.M.; Treimo, J.; Narvhus, J.A. Effect of 0302(98)75620-6.
temperature on growth and metabolism of 54. Hashim, I.B.; Khalil, A.H.; Afifi, H.S. Quality
probiotic bacteria in milk. International Dairy characteristics and consumer acceptance of
Journal 2005, 15, 989-997, yogurt fortified with date fiber. Journal of Dairy
doi:https://doi.org/10.1016/j.idairyj.2004.08.015 Science 2009, 92, 5403-5407,
44. Billard, I.; Mekki, S.; Ouadi, A.; Gaillard, C.; doi:https://doi.org/10.3168/jds.2009-2234.
Trubert, D.; Moutiers, G. TTA solvation kinetics 55. Damin, M.R.; Alcântara, M.R.; Nunes, A.P.;
in the ionic liquid BumimTf2N. Comptes Rendus Oliveira, M.N. Effects of milk supplementation
Chimie 2007, 10, 1117-1121, with skim milk powder, whey protein
doi:https://doi.org/10.1016/j.crci.2007.04.007. concentrate and sodium caseinate on
45. Eissa, E.A.; Ahmed, I.M.; Yagoub, A.E.A.; acidification kinetics, rheological properties and
Babiker, E.E. Physicochemical, microbiological structure of nonfat stirred yogurt. LWT - Food
and sensory characteristics of yoghurt produced Science and Technology 2009, 42, 1744-1750,
from goat milk. Livestock Research for Rural doi:https://doi.org/10.1016/j.lwt.2009.03.019.
Development 2010, 22, 247-253. 56. Shah, N.P. Effects of milk-derived bioactives: an
46. Demirci, T.; Aktaş, K.; Sözeri, D.; Öztürk, H.İ.; overview. British Journal of Nutrition 2000, 84,
Akın, N. Rice bran improve probiotic viability in 3-10, doi:10.1017/S000711450000218X.
yoghurt and provide added antioxidative 57. Blum, U. Effects of Microbial Utilization of
benefits. Journal of Functional Foods 2017, 36, Phenolic Acids and their Phenolic Acid
396-403, Breakdown Products on Allelopathic
doi:https://doi.org/10.1016/j.jff.2017.07.019. Interactions. Journal of Chemical Ecology 1998,
47. Hu, G.; Yu, W. Effect of hemicellulose from rice 24, 685-708, doi:10.1023/A:1022394203540.
bran on low fat meatballs chemical and 58. Ghasemzadeh, A.; Karbalaii, M.T.; Jaafar,
functional properties. Food Chemistry 2015, 186, H.Z.E.; Rahmat, A. Phytochemical constituents,
239-243. antioxidant activity, and antiproliferative
48. Ozcan, T.; Kurtuldu, O. Influence of dietary fiber properties of black, red, and brown rice bran.
addition on the properties of probiotic yogurt. Chemistry Central Journal 2018, 12, 17,
International Journal of Chemical Engineering doi:10.1186/s13065-018-0382-9.
and Applications 2014, 5, 397. 59. Thompson, J.L.; Lopetcharat, K.; Drake, M.A.
49. El-Said, M.M.; Haggag, H.F.; Fakhr El-Din, Preferences for Commercial Strawberry
H.M.; Gad, A.S.; Farahat, A.M. Antioxidant Drinkable Yogurts Among African American,
activities and physical properties of stirred Caucasian, and Hispanic Consumers in the
yoghurt fortified with pomegranate peel extracts. United States. Journal of Dairy Science 2007, 90,
Annals of Agricultural Sciences 2014, 59, 207- 4974-4987,
212, doi:https://doi.org/10.3168/jds.2007-0313.
doi:https://doi.org/10.1016/j.aoas.2014.11.007. 60. Ouml; nder, Y.; Sadiye, P.E. Functional
50. Donkor, O.N.; Nilmini, S.L.I.; Stolic, P.; components of berry fruits and their usage in
Vasiljevic, T.; Shah, N.P. Survival and activity of food technologies. African Journal of
selected probiotic organisms in set-type yoghurt Agricultural Research 2009, 4, 422-426.
during cold storage. International Dairy Journal 61. Yuksel, Z.; Avci, E.; Erdem, Y.K.
2007, 17, 657-665, Characterization of binding interactions between
doi:https://doi.org/10.1016/j.idairyj.2006.08.006 green tea flavanoids and milk proteins. Food
51. Grigelmo-Miguel, N.; Gorinstein, S.; Martı́n- Chemistry 2010, 121, 450-456,
Belloso, O. Characterisation of peach dietary doi:https://doi.org/10.1016/j.foodchem.2009.12.
fibre concentrate as a food ingredient. Food 064.
Chemistry 1999, 65, 175-181, 62. Massey, L.K. Dairy Food Consumption, Blood
doi:https://doi.org/10.1016/S0308- Pressure and Stroke. The Journal of Nutrition
8146(98)00190-3.
________________________________________________
Egypt. J. Chem. 66, No. 12 (2023)
 Angiotensin-converting enzyme (ACE-1) inhibitory and antioxidant activities of probiotic yogurt enriched with rice bran 365
__________________________________________________________________________________________________________________

2001, 131, 1875-1878, media for enumeration of probiotic micro-

doi:10.1093/jn/131.7.1875. organisms in combination with yogurt starter
63. Shahidi, F.; Zhong, Y. Bioactive Peptides. cultures in fermented milk. African Journal of
Journal of AOAC INTERNATIONAL 2008, 91, Microbiology Research 2011, 5, 3901-3906.
914-931, doi:10.1093/jaoac/91.4.914. 69. Senaka Ranadheera, C.; Evans, C.A.; Adams,
64. Ashraf, R.; Shah, N.P. Selective and differential M.C.; Baines, S.K. Probiotic viability and
enumerations of Lactobacillus delbrueckii subsp. physico-chemical and sensory properties of plain
bulgaricus, Streptococcus thermophilus, and stirred fruit yogurts made from goat’s milk.
Lactobacillus acidophilus, Lactobacillus casei Food Chemistry 2012, 135, 1411-1418,
and Bifidobacterium spp. in yoghurt — A doi:https://doi.org/10.1016/j.foodchem.2012.06.
review. International Journal of Food 025.
Microbiology 2011, 149, 194-208, 70. do Espírito Santo, A.P.; Cartolano, N.S.; Silva,
doi:https://doi.org/10.1016/j.ijfoodmicro.2011.0 T.F.; Soares, F.A.S.M.; Gioielli, L.A.; Perego,
7.008. P.; Converti, A.; Oliveira, M.N. Fibers from fruit
65. Rezaei, A.; Amirdivani, S.; Asl, A.K.; by-products enhance probiotic viability and fatty
Malekinejad, H.; Zomorodi, S.; Hosseinmardi, F. acid profile and increase CLA content in
Inhibition of the Angiotensin I Converting yoghurts. International Journal of Food
Enzyme (ACE) and proteolysis of non-fat Microbiology 2012, 154, 135-144,
probiotic yogurt. Brazilian Journal of Food doi:https://doi.org/10.1016/j.ijfoodmicro.2011.1
Technology 2019, 22. 2.025.
66. Donkor, O.N.; Henriksson, A.; Singh, T.K.; 71. Sendra, E.; Fayos, P.; Lario, Y.; Fernández-
Vasiljevic, T.; Shah, N.P. ACE-inhibitory López, J.; Sayas-Barberá, E.; Pérez-Alvarez, J.A.
activity of probiotic yoghurt. International Dairy Incorporation of citrus fibers in fermented milk
Journal 2007, 17, 1321-1331, containing probiotic bacteria. Food
doi:https://doi.org/10.1016/j.idairyj.2007.02.009 Microbiology 2008, 25, 13-21,
. doi:https://doi.org/10.1016/j.fm.2007.09.003.
67. Espírito Santo, A.P.d.; Silva, R.C.; Soares, 72. Santos, R.O.; Silva, M.V.F.; Nascimento, K.O.;
F.A.S.M.; Anjos, D.; Gioielli, L.A.; Oliveira, Batista, A.L.D.; Moraes, J.; Andrade, M.M.;
M.N. Açai pulp addition improves fatty acid Andrade, L.G.Z.S.; Khosravi-Darani, K.; Freitas,
profile and probiotic viability in yoghurt. M.Q.; Raices, R.S.L.; et al. Prebiotic flours in
International Dairy Journal 2010, 20, 415-422, dairy food processing: Technological and
doi:https://doi.org/10.1016/j.idairyj.2010.01.002 sensory implications. International Journal of
. Dairy Technology 2018, 71, 1-10,
68. Saccaro, D.M.; Hirota, C.Y.; Tamime, A.Y.; de doi:https://doi.org/10.1111/1471-0307.12394.
Oliveira, M.N. Evaluation of different selective

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Egypt. J. Chem. 66, No. 12 (2023)