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Isolation and Identification of Lactic Acid Bacteria from
Fermented Soybean Pulp and Its Application as Starter Culture for
Soy Yogurt
Charity Joy L. Sumagaysay
1
*, James B. Sumagaysay
1
, Jeany Mae H. Menisterio
2
, Lalaine Grace M. Robles
3
, Reggie Y.
Dela Cruz
3
, and Zeus S. Elumba
3
1
Mountain View College, College Heights, Mt. Nebo, Valencia City, Bukidnon, Philippines
2
Research and Development, East-West Seed Company Incorporated, Buena Vista Farm, Zone 5, Sankanan Manolo
Fortich, Bukidnon, Philippines
3
Institute of Biological Sciences, Central Mindanao University, Maramag, Bukidnon, Philippines
DOI: https://doi.org/10.51583/IJLTEMAS.2025.1407000107
Received: 17 July 2025; Accepted: 24 July 2025; Published: 14 August 2025
Food waste processing holds great potential in mitigating global food insecurity. Valorization of food waste is a sustainable practice
that also lessens the effects of climate change. Isolation, identification, and characterization of lactic acid bacteria from fermented
soya pulp was conducted. The isolated lactic acid bacteria species was utilized as starter culture to produce soy yogurt. This study
was performed to identify LAB isolated from fermented soya pulp using 16s rRNA gene sequencing; to ferment soymilk using the
LAB isolated from fermented soya pulp and develop soy yogurt; to determine the proximate nutritional composition of the soy
yogurt through proximate analysis; and to evaluate the sensory characteristics of the yogurt such as color, aroma, texture, taste, and
overall acceptability. The 16s rRNA gene sequencing procedure identified the isolated lactic acid bacteria species as Pediococcus
pentosaceus Mees, 1934. Four yogurt samples were formulated using P. pentosaceus and a commercial starter culture. Proximate
compositions such as titratable acidity, ash content, crude protein, and crude fat were determined. The soy-based yogurt drinks were
similarly high in protein compared with the cow’s milk counterpart. Notably, formulation 1 with the P. pentosaceus as the sole
starter culture scored highest in crude protein. Significant difference in the average ash content between the four samples was
observed. Post-hoc analyses using the Bonferroni correction revealed that formulation 1 with the isolated starter culture, P.
pentosaceus, had the highest ash content. Sensory evaluation revealed that formulation 2 with both the isolated starter culture and
commercial starter gained peak in color, taste, and overall acceptability. There was significant difference in the average aroma and
texture scores between the four samples, favoring the cow’s milk yogurt sample for the aroma and the soymilk yogurt with both P.
pentosaceus and commercial starter culture for the texture. This study showed the potential of P. pentosaceus as a sole starter
culture and in combination with commercial starter culture in producing functional foods from food waste such as soymilk-based
yogurt. Refinement and standardization of the yogurt production protocol is recommended to produce the yogurt with better texture
and taste. By so doing, product development from food waste can support sustainable food production and mitigate global food
insecurity and climate change.
Keywords: isolation, identification, lactic acid bacteria, starter culture, soymilk yogurt, okara
I. Introduction
Global food insecurity is on the rise these days. According to the Food and Agriculture Organization (FAO), around 1.3 billion tons
of food made for human consumption is wasted annually. This number is equivalent to more than one-third of the total food
produced worldwide. These deficits would result to wastage of resources (FAO, 2019). One solution to this pressing problem is
valorization of food waste. Food waste processing methods include isolating, characterizing, and identifying lactic acid bacteria to
produce fermented functional food products.
Meanwhile, most of the soybean pulp products are outcomes of fermentation using various microorganisms. Fermented foods have
better shelf life, bioactive molecules, vitamins, and enhanced availability of necessary contents (Rezac et al., 2018). Some also have
therapeutic properties such as improving gut microbiota and gastrointestinal health and overcoming diabetes and cardiovascular
diseases (Melini et al., 2019). In a study in 2024, the soya pulp was used as a culture medium for lactic acid bacteria to minimize
improper disposal of soymilk by-product and to explore soya pulp potential as a culture medium to grow lactic acid bacteria and
save the cost of buying the standard MRS medium (Retnowati et al., 2024).
Lactic acid bacteria have been reported to possess several health benefits such as improving gut health, modulating appetite, and
managing weight, modulating the immune system, decreasing the severity of infection, showing antimicrobial effects on food-borne
pathogens, lowering blood pressure, high radical scavenging activity, and improving cognitive health (Mathur et al., 2020). Because
they are generally recognized as safe (GRAS), they are considered as one of the most important groups of microorganisms being
utilized as starter cultures in producing functional food, exerting probiotic effects, and exhibiting food preservation potential (Frick
et al., 2007). Due to their fermentative nature, they are used to produce dairy, meat, and vegetables fermented products (De Vuyst
and Leroy, 2007). Lactic acid bacteria isolates can be identified from fermented foods which are being studied for the potential of
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antimicrobial activity against foodborne pathogens and non-resistance to antibiotics (Banik et al., 2023). By far, dairy, and
fermented foods provide the main source of probiotics. But because of lactose intolerance and high cholesterol levels, non-dairy
fermented products are now gaining more interests (Liu et al., 2022). Lactic acid bacteria can be used against food-borne pathogens
using fermentates. Food-grade cultures and undefined fermentates of lactic acid bacteria are more consumer-friendly and do not
require extensive labelling, in contrast to purified fermentates which require extensive labelling as additives (Field et al., 2015).
At present, the usefulness of 16S rRNA gene sequence is expanded to food processing application where molecular characterization
has recently become essential in classifying and identifying lactic acid bacteria (Ayivi and Ibrahim, 2022). This modern step of
molecular identification for lactic acid bacteria isolated from soya milk or soya pulp can be done through 16S rRNA sequencing
(Retnowati et al., 2024).
Several of studies have been published about yogurt production using lactic acid bacteria and cow’s milk. However, in soymilk
yogurt production, only very few research is available. Limited reviews mention only a few isolated lactic acid bacteria from
fermented soybean pulp; mostly are from different fermented sources (Qi et al., 2021; Jiang et al., 2021). Of the few isolated lactic
acid bacteria from fermented soybean pulp, P. pentosaceus was only used as a mixed starter culture for soymilk yogurt along with
other lactic acid bacteria species. There are no published studies on using P. pentosaceus isolated from fermented soybean pulp as
a pure starter culture for soymilk yogurt production. Hence, this study was conceived.
II. Materials and Methods
Place and Duration of Study
The molecular characterization of lactic acid bacteria samples was done by Macrogen Inc., 1007, 254 Beotkkot-ro, Geuncheon-gu,
Seoul 08511, Rep. of Korea. The preparation of soy yogurt was conducted at the Isolation Room of the Department of Biology in
Mountain View College of Seventh-day Adventists, Inc., College Heights, Mt. Nebo, Valencia City. Lyophilization or freeze-drying
of the samples and proximate analysis were conducted at the Natural Science Research Institute (NSRI) and Center for Food
Research, Innovation, and Extension (CFRIE) of Central Mindanao University, University Town, Musuan, Maramag, Bukidnon,
Philippines. Sensory evaluation was performed by selected participants. The study started on February 5, 2025 until May 2025.
Molecular Characterization of LAB Isolated from Fermented Soya Pulp
Pure lactic acid bacteria that were previously isolated from fermented soya pulp and stored in 85% MRS broth and 15% Glycerol
were sent to Kinovett Scientific Solutions, Co. Then, online request for services to Macrogen Inc. was made following the
procedures prescribed by Kinovett Scientific Solutions, Co. The service requested was identification, specifically 16S rRNA gene
sequencing. Kinovett Scientific Solutions, Co. facilitated the air mail of the samples to Macrogen Inc., a South Korean public
biotechnology company. Macrogen communicated the progress, results, and payment invoice through emails.
Preparation and Allocation of Starter Culture
The method of Utami et al., (2020) was followed with some modifications. Frozen stock cultures of isolated lactic acid bacteria
from fermented soya pulp was thawed and activated in 1 L 10% skim milk and 1% sucrose mixture. The mixture was incubated at
30°C for 18 hours. The 1L starter culture was allocated into three different formulations: each formulation received 100 mL of the
starter culture per replicate. There were three replicates per formulation.
Preparation of Soy Yogurt
For this procedure, the method of Ugwona et al., (2018) was followed with some modifications. The following set-ups were made:
Control (Soymilk + Yakult), Formulation 1 (Soymilk + Starter Culture), Formulation 2 (Soymilk + Yakult + Starter Culture),
Formulation 3 (Cow’s milk + Starter Culture). Three replicates were made per set-up. One point five liter (1.5 L) of soymilk was
used in all soymilk set-ups. The Cow’s milk was prepared by mixing 125 g of full cream powdered milk (Bear Brand) with 1 L of
distilled water. The liquid Cow’s milk and soymilk samples were separately pasteurized at 85°C for 15 mins and cooled at 44°C.
After pasteurization, formulations 1-3 were inoculated with 100 mL (6.7%) starter culture per replicate at 44°C. Four (4) bottles of
Yakult (80 mL per bottle) with live Lacticaseibacillus paracasei strain Shirota was used for each liter of soymilk in the control
sample and formulation 2 sample. One-hundred milliliters (100 mL) of starter culture was mixed with soymilk in formulation 1 and
another 100 mL of starter culture was added into the soymilk and Yakult mixture in formulation 2. One-hundred milliliters (100
mL) of starter culture was also mixed with Cow’s milk in formulation 3. All the samples were fermented in a water bath at 44°C
for 7h and then allowed to cool gradually.
Allocation of Samples for Proximate Analysis and Nutrition Facts Analysis
Fresh soy yogurt samples were brought to the Center for Food Research, Innovation, and Extension (CFRIE) of Central Mindanao
University for immediate analysis of titratable acidity and ash content. Freeze-drying at -86 °C for 42 hours was performed using
the lyophilizer at the Tuklas Lunas Development Center (TLDC). Freeze drying was done for the analysis of percentage crude fat
and crude protein.
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Sensory Evaluation
Ten (10) semi-trained panelists performed the sensory evaluation assessing the attributes of the yogurt such as: color, aroma, texture,
taste, and overall acceptability. Their feedback was recorded using a hedonic scale, ranging from 1 (extremely disliked) to 9
(extremely liked). Labels of the finish product were concealed from the panelists to prevent bias (Asghar et al., 2022).
Statistical Analysis
Data were analyzed using Microsoft Excel (2016). Analysis of Variance (ANOVA) was used to compare means at p < 0.05 level
of significance.
III. Results and Discussion
Identification of the LAB Isolated from Fermented Soya Pulp through 16s rRNA Sequencing
Macrogen Inc. identified the lactic acid bacteria species that was isolated from the fermented soya pulp as Pediococcus pentosaceus
Mees, 1934 (Figure 1). Pediococcus is a genus of Gram-positive lactic acid bacteria, previously placed within the family of
Streptococcaceae (Chen et al., 2020), and recently classified under the family of Lactobacillaceae (d’Acierno et al., 2025). P.
pentosaceus is one of the 12 species of Pediococcus that have been described; and along with P. acidilactici, are associated with
dairy products such as cheese, fermentation of milk, and cheese manufacture (Holland et al., 2010). Pediococcus species usually
occur in pairs or tetrads, and divide along two planes of symmetry. They are coccus shaped microbes, non-motile, and non-spore
forming. The end product of their metabolism is lactic acid, making them acid tolerant, and thus classifying them under lactic acid
bacteria. They are also responsible for the fermentation of cabbage, making it sauerkraut, together with Leuconostoc and
Lactobacillus (Macrogen, 2025). Previous studies confirmed that P. pentosaceus occurs naturally in spontaneously fermented
products that could have a considerable role in quality, safety, and efficiency when put together with other bacteria in a mixed
fermenter (Gong and Qi, 2020; Montemurro et al., 2020; Xu et al., 2021). Like most lactic acid bacteria, P. pentosaceus are
anaerobic and ferment sugars. They can be found in plant materials, ripened cheese, and a variety of processed meats (Macrogen,
2025).
Figure 1. Molecular identification of the isolated lactic acid bacteria from fermented soya pulp using 16S rRNA gene sequencing.
a) Sample from moist colony. b) Sample from mucoid colony. c) Sample from round colony. d) Sample from
irregularly shape colony.
Pediococcus pentosaceus has many health benefits such as probiotic effects, rich in antioxidants, lowering cholesterol, and boosting
the immune system. Recently, P. pentosaceus’ potential as an ingredient in probiotic fermentation products increasingly attracts
more attention (Qi et al., 2021). A study in 2022 stated that P. pentosaceus is increasingly studied in the dairy industry due to its
promising qualities as a starter culture bacterium. The study explored the physiological, survivability, and genomic properties of P.
pentosaceus. The strain revealed amazing tolerance in low pH and high concentrations of bile salts. It also showed 71% and 84%
activity against pathogens, strong biofilm formation, and notable resistance to antibiotics. The whole genome sequence also showed
various genes responsible for stress tolerance, adhesion, biofilm formation, as well as acids and bile salts tolerance (Mgomi et al.,
2022).
The use of P. pentosaceus from different sources is widespread in several applications such as antioxidant and fatty acid profiling
of fermented goat, camel, and cow’s milk (Balakrishnan and Agrawal, 2014); as an adjuvant therapeutic option for patients with
ulcerative colitis (UC) (Bamba et al., 2018); anti-listerial activity (Jang et al., 2014); as starter culture in sourdough bread-making
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0.73 0.5 0.61 0.43
5.63
6.13
5.89
3.37
15.4
19.63
18.8
15.45
1.73
7.61
4.59
15.02
0
5
10
15
20
25
Control F1 F2 F3
Percentage (%)
Titratable Acidity Ash Content Crude Protein Crude Fat
(Plessas et al., 2020); as a heat stable, bacteriocin-producing and vancomycin-sensitive lactic culture from beans (Venkateshwari
et al., 2010); and producing key odorants and non-volatile metabolites in broccoli juices (Xu et al., 2021). However, in soymilk
yogurt production, only very few researches have been published; and those studies only mention the use of P. pentosaceus as a
mixed starter culture for soymilk yogurt along with other lactic acid bacteria species. In the two reviews read about P. pentosaceus,
there was no mention about it being isolated from the fermented soybean pulp, but from other different fermented sources (Qi et
al., 2021; Jiang et al., 2021). There are also no published studies on using P. pentosaceus isolated from fermented soybean pulp as
a pure starter culture for soymilk yogurt production. Hence, this study was conceived.
Proximate Nutritional Composition of the Soy Yogurt
There were four proximate compositions determined by the Center for Food Research, Innovation, and Extension (CFRIE) of the
Central Mindanao University. One was a physico-chemical component called, titratable acidity; and chemical components such as
ash, crude protein, and crude fat. All were determined in percentage form (Table 1).
Table 1. Mean percentage and standard deviation values of the proximate nutritional compositions of the soy yogurt drink
samples.
Proximate Composition
Control (%)
F1 (%)
F2 (%)
F3 (%)
Titratable Acidity
0.73±0.009
0.61±0.02
Ash Content
5.63±0.08
a
5.89±0.82
f
Crude Protein
15.4±0.25
18.8±0.73
Crude Fat
1.73±0.08
4.59±0.22
*mean value of the same letter are not significantly different at P value 0.05.
The titratable acidity, crude protein, and crude fat of the samples were comparably similar. The soymilk based samples however
showed high fermentation activity indicated in the titratable acidity. This meant that soymilk based samples produced more lactic
acid compared to the control. Meanwhile, the soy-based yogurt drinks were similarly high in protein compared with the cow’s milk
counterpart. Notably, formulation 1 with the P. pentosaceus as the sole starter culture scored highest in crude protein. This meant
that soy-based yogurts can be good protein source alternatives for cow’s milk yogurt. Additionally, the results revealed that P.
pentosaceus can be used solely as a starter culture to develop equally protein rich, plant-based alternative functional food from food
waste.
There was significant difference in the average ash content between the four samples. Post-hoc analyses using the Bonferroni
correction revealed that the amount of ash in the control group scored significantly lower compared with the ash of formulation 1.
The ash of the control group scored significantly higher than formulation 3. The ash of formulation 1 also scored significantly
higher than formulation 3; and the ash of formulation 2 also scored significantly higher than formulation 3. Notably, formulation
1 with the P. pentosaceus as the sole starter culture scored highest in the ash content. This may suggest higher mineral content that
is naturally present in formulation 1 (Figure 2).
Figure 2. Proximate nutritional components of the soymilk-based yogurt drink samples: control, formulation 1, and formulation 2;
in comparison with formulation 3 as the cow’s milk-based yogurt drink counterpart. All the results are presented as
mean ± standard deviation of the mean (n=3).
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Sensory Characteristics of the Yogurt
A 9-point hedonic scale sensory evaluation of the formulated yogurt drink samples was used. There were five sensory parameters
considered: color, aroma, texture, taste, and overall acceptability (Table 2).
The color, taste, and overall acceptability of the yogurt drink samples were comparably similar. However, formulation 2 (P.
pentosaceus with commercial starter culture) scored highest in color, taste, and overall acceptability. Although using different lactic
acid bacteria species, this result is similar with Olusola and Adepoju (2022), where the soy yogurt fermented by both the isolated
lactic acid bacteria starter culture and the commercial starter culture scored higher in sensory evaluation in contrast to the soy yogurt
solely fermented by the isolated lactic acid bacteria.
Table 2. Mean and standard deviation values of the proximate nutritional compositions of the soy yogurt drink samples.
Sensory Parameters
Control
F1
F2
F3
Color
6.5±0.97
6.5±1.18
7.3±1.16
6.6±1.65
Aroma
6.9±0.74
a
5.5±0.97
b
6.8±1.55
a
7±1.25
a
Texture
5.2±0.42
a
6.1±1.29
a
6.3±1.06
b
5±0.82
c
Taste
6.3±0.67
6±0.47
7.5±0.53
4.6±0.52
Over-All Acceptability
7.1±0.88
5.8±0.79
7.2±0.79
3.1±1.10
There was significant difference in the average aroma scores between the four samples. Post-hoc analyses using the Bonferroni
correction revealed that the aroma of the control group scored significantly higher compared with the aroma of formulation 1
(Tables 3 and 4).
Table 3. Mean and variance values of the aroma scores in the yogurt drink samples.
Table 4. ANOVA of the aroma scores in the yogurt drink samples.
Source of Variation
SS
df
MS
F
P-value
F crit
Between Groups
14.9
3
4.966667
3.64898
0.02143
2.86626555
Within Groups
49
36
1.361111
Total
63.9
39
There was also significant difference in the average texture scores between the four samples. Post-hoc analyses using the Bonferroni
correction revealed that the texture of the control group scored significantly lower compared with the texture of formulation 2; and
the texture of formulation 2 scored significantly higher than formulation 3 (Tables 5 and 6).
Table 5. Mean and ANOVA values of the texture scores in the yogurt drink samples.
Groups
Count
Sum
Average
Variance
Control
10
52
5.2
0.177778
T1
10
61
6.1
1.655556
T2
10
63
6.3
1.122222
T3
10
50
5
0.666667
Groups
Count
Sum
Average
Variance
Control
10
69
6.9
0.544444
F1
10
55
5.5
0.944444
F2
10
68
6.8
2.4
F3
10
70
7
1.555556
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Table 6. Mean and ANOVA values of the texture scores in the yogurt drink samples.
Source of Variation
SS
df
MS
F
P-value
F crit
Between Groups
12.5
3
4.166667
4.601227
0.00795
2.866266
Within Groups
32.6
36
0.905556
Total
45.1
39
Over-all, there were significant differences in the aroma and texture of the yogurt drink samples. Formulation 1 (P. pentosaceus)
scored lowest in odor, while formulation 3 (cow’s milk) revealed the most favored aroma. The lower preference in aroma for soy-
based yogurt may be caused by its naturally, slightly beany or grassy smell. Meanwhile, formulation 2 (P. pentosaceus and
commercial starter culture) scored highest in texture followed by formulation 1 (P. pentosaceus) (Figure 3). This result is similar
with Olusola and Adepoju (2022).
Figure 3. Sensory evaluation on attributes: color, aroma, texture, taste, and over-all acceptability of soymilk-based and cow’s
milk-based yogurt drinks. All the results are presented as mean ± standard deviation of the mean (n=10).
IV. Conclusion and Recommendation
Valorization of food waste using the naturally fermented soybean pulp as the prebiotic ingredient in isolating lactic acid bacteria
with a probiotic potential was conducted prior to this study. Results revealed that the isolated lactic acid bacterium was identified
as Pediococcus pentosaceus Mees, 1934. This species was utilized as a starter culture in producing the soymilk yogurt drinks in
this study. Proximate analysis showed that formulation 1 with the P. pentosaceus as the sole starter culture scored highest in crude
protein and ash content. It also scored second highest in crude fat. There was significant difference in the average ash content
between the four samples. The color, taste, and overall acceptability of the yogurt drink samples were comparably similar. But
formulation 2 scored highest in color, taste, and overall acceptability. Significant differences were observed in the average aroma
and textures scores.
This study showed the potential of P. pentosaceus as a sole starter culture and in combination with commercial starter culture in
producing functional foods from food waste such as soymilk-based yogurt. Refinement of the methods and ingredients used in the
process is recommended to produce the yogurt with the desired texture and taste. It is also recommended to include the
determination of moisture, pH, total soluble solids for the sweetness, and further nutritional facts analysis of the product. Further,
increasing the participants for the sensory analysis is also recommended to improve the product’s overall acceptability for
commercial production. Bile tolerance test and antimicrobial screening against foodborne pathogens and non-resistance to
antibiotics are also suggested to explore more of this species characteristics and usability. By so doing, product development from
food waste can support sustainable food production and mitigate global food insecurity and climate change.
References
1. Aritonang, Salam N., Elly Roza, Evy Rossi, Endang Purwati and Husmaini. (2017). Isolation and identification of lactic
acid bacteria from okara and evaluation of their potential as candidate probiotics. Pak. J. Nutr., 16: 618-628.
2. Asghar, Aasma; Afzaal, Muhammad; Nosheen, Farhana; Saeed, Farhan; Nayik, Gulzar Ahmad; AL-Farga, Ammar;
Alansari, Wafa S.; Eskandrani, Areej A.; and Shamlan, Ghalia. (2022). Isolation and Molecular Characterization of
6.5
6.9
5.2
6.3
7.1
6.5
5.5
6.1
6
5.8
7.3
6.8
6.3
7.5
7.2
6.6
7
5
4.6
3.1
0
1
2
3
4
5
6
7
8
Color Aroma Texture Taste Over-All
Acceptability
Hedonic Scale, 1
-9
Control F1 F2 F3
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www.ijltemas.in Page 905
Processed Soybean Waste for the Development of Synbiotic Yogurt. Fermentation, 8, 622.
https://doi.org/10.3390/fermentation8110622.
3. Ayivi, R. D., Gyawali, R., Krastanov, A., Aljaloud, S. O., Worku, M., Tahergorabi, R., Silva, R. C. d., & Ibrahim, S. A.
(2020). Lactic acid bacteria: Food safety and human health applications. Dairy, 1(3), 202–232.
https://doi.org/10.3390/dairy1030015
4. Ayivi, Raphael D. and Ibrahim, Salam A. (2022). Lactic acid bacteria: an essential probiotic and starter culture for the
production of yoghurt. International Journal of Food Science and Technology, 57:7008–7025.
5. Axelsson, L. (2004). Lactic acid bacteria: classification and physiology. Food Science and Technology-New York-Marcel
Dekker. 139:1-66.
6. Balakrishnan, G., and Agrawal, R. (2014). Antioxidant activity and fatty acid profile of fermented milk prepared by
Pediococcus pentosaceus. J. Food Sci. Technol. 51, 4138–4142. doi: 10.1007/s13197-012-0891-9
7. Bamba, S., Takahashi, K., Imaeda, H., Nishida, A., Kawahara, M., Inatomi, O., et al. (2018). Effect of fermented vegetable
beverage containing Pediococcus pentosaceus in patients with mild to moderate ulcerative colitis. Biomed. Rep. 9, 74–80.
doi: 10.3892/br.2018.1099
8. Banik, A., Anjum, H., Habib, H., Abony, M, Begum, A., Ahmed, Z. (2023). Characterization of lactic acid bacteria
isolated from street pickles of Dhaka, Bangladesh. Heliyon 9 e17508.
9. Bintsis, Thomas. (2018). Lactic acid bacteria as starter cultures : An update in their metabolism and genetics. AIMS
Microbiology, 4(4): 665–684. DOI: 10.3934/microbiol.2018.4.665
10. Bren d’Amour, C., Reitsma, F., Baiocchi, G., Barthel, S., Güneralp, B., Erb, K.-H., . . . Seto, K. C. (2017). Future urban
land expansion and implications for global croplands. Proceedings of the National Academy of Sciences, 114(34), 8939.
doi:http://doi.org/10.1073/pnas.1606036114
11. Chen, Y. S., F. Yangida & T. Shinahara. (2005). Isolation and identification of lactic acid bacteria from soil using an
enrichment procedure. Lett. Applied Microbiol., 40: 195-200.
12. Chen, T., Wang, L., Li, Q., Long, Y., Lin, Y., Yin, J., et al. (2020). Functional probiotics of lactic acid bacteria from Hu
sheep milk. BMC Microbiol. 20:228. doi: 10.1186/s12866-020-01920-6
13. Cirlini, M., Ricci, A., Galaverna, G., & Lazzi, C. (2020). Application of lactic acid fermentation to elderberry juice:
Changes in acidic and glucidic fractions. LWT, 118, 108779.
14. d’Acierno A, Nazzaro F, Reale A, Sorrentino A, Rossi M, Bucci G, Ianigro M. (2025). CNR-ISA Agri-food
Microorganism Dataset (ISA01-CC). Version 1.1. Consiglio Nazionale delle Ricerche, Istituto di Bioscienze e BioRisorse
(CNR-IBBR). Occurrence dataset https://doi.org/10.15468/2ze5cv accessed via GBIF.org on 2025-04-20.
https://gbif.org/occurrence/4920304437
15. Daggett, P-M. and Simione, FP. (1989). Method of culturing freeze-dried microorganisms and resultant preparation.
US4879239A United States.
16. Dash, Chinmaya & Payyappilli, Rajan. (2016). KOH string and Vancomycin susceptibility test as an alternative method
to Gram staining. Journal of International Medicine and Dentistry. 3. 88-90. 10.18320/JIMD/201603.0288.
17. Dawe, D., Dobermann, A., Moya, P., Abdulrachman, S., Singh, B., Lal, P., . . . Zhen, Q. X. (2000). How widespread are
yield declines in long-term rice experiments in Asia? Field Crops Research, 66(2), 175-193.
doi:https://doi.org/10.1016/S0378-4290(00)00075-7.
18. De Vuyst L & Leroy F. (2007). Bacteriocins from Lactic Acid Bacteria: Production, Purification, and Food Applications.
J Mol Microbiol Biotechnol. 13: 194-199. FAO. (2019). Food Loss and Food Waste. Retrieved from
http://www.fao.org/foodloss-and-food-waste/en/
19. Elida, M., A. Agustina, E. Ermiati, and S. Desminarte. (2022). Isolate Characterization and Amylolytic Properties of Lactic
Acid Bacteria from Traditional Fermented Dadih. 1st Lekantara Annual Conference on Natural Science and Environment
(LeNS 2021). IOP Conf. Series: Earth and Environmental Science 1097 (2022) 012025. IOP Publishing.
doi:10.1088/1755-1315/1097/1/01202.
20. Field, D.; Daly, K.; O’Connor, P.M.; Cotter, P.D.; Hill, C.; Ross, R.P. (2015). Efficacies of Nisin A and Nisin V Semi-
purified Preparations Alone and in Combination with Plant Essential Oils for Controlling Listeria monocytogenes. Appl.
Environ. Microbiol. 2015, 81, 2762–2769.
21. Filannino, P., Di Cagno, R., & Gobbetti, M. (2018). Metabolic and functional paths of lactic acid bacteria in plant foods:
Get out of the labyrinth. Current Opinion in Biotechnology, 49, 64–72.
22. Frick, Julia-Stefanie, Katrin Schenk, Matteo Quitadamo, Frauke Kahl, Martin Köberle, Erwin Bohn, Martin Aepfelbacher,
& Ingo B. Autenrieth. (2007). Lactobacillus fermentum attenuates the proinflammatory effect of Yersinia enterocolitica
on human epithelial cells. Inflammatory Bowel Diseases, 13(1): 83-90.
23. Farmer, S. and Alibek, K. (2019). Production and preservation of bacillus reference culture for generating standardized
and reliable inocula. WO2019222168A1 WIPO (PCT).
24. Gong, Y., and Qi, X. (2020). A study revealing volatile aroma produced by Pediococcus pentosaceus in dough
fermentation. Food Sci. Nutr. 8, 5077–5085. doi: 10.1002/fsn3.1807
25. Gobbetti, M., M. De Angelis, R. Di Cagno, L. Mancini, and P. F. Fox. (2015). Pros and cons for using non-starter lactic
acid bacteria (NSLAB) as secondary/adjunct starters for cheese ripening. Trends Food Sci. Technol. 45:167–178. https: /
/ doi .org/ 10 .1016/ j.tifs .2015 .07 .016.
INTERNATIONAL JOURNAL OF LATEST TECHNOLOGY IN ENGINEERING,
MANAGEMENT & APPLIED SCIENCE (IJLTEMAS)
ISSN 2278-2540 | DOI: 10.51583/IJLTEMAS | Volume XIV, Issue VII, July 2025
www.ijltemas.in Page 906
26. Gudisa, A. and Yenasew, A. (2022). Isolation, Identification, and Biochemical Characterization of Lactic Acid Bacteria
from Okara. Food Science and Quality Management. www.iiste.org ISSN 2224-6088 (Paper) ISSN 2225-0557 (Online)
Vol.117, 2022
27. Holland, R., Crow, V., & Curry, B. (2010). Lactic Acid Bacteria | Pediococcus spp. Encyclopedia of Dairy Sciences
(Second Edition), 149-152. https://doi.org/10.1016/B978-0-12-374407-4.00269-7
28. Hu, Dan; Wu, Jinyong; Jin, Long; Yuan, Lixia; Li, Jun; Chen, Xiangsong; and Yao, Jianming. (2021). Evaluation of
Pediococcus pentosaceus strains as probiotic adjunct cultures for soybean milk post-fermentation. Food Research
International, Volume 148, 2021, 110570, ISSN 0963-9969, https://doi.org/10.1016/j.foodres.2021.110570.
(https://www.sciencedirect.com/science/article/pii/S0963996921004695)
29. Jang, S., Lee, J., Jung, U., Choi, H. S., and Suh, H. J. (2014). Identification of an anti-listerial domain from Pediococcus
pentosaceus T1 derived from Kimchi, a traditional fermented vegetable. Food Control 43, 42–48.
doi:10.1016/j.foodcont.2014.02.040
30. Jenkins, Claire; Ling, Clare L.; Ciesielczuk, Holly L.; Lockwood, Julianne; Hopkins, Susan; McHugh, Timothy D.;
Gillespie, Stephen H.; and Kibbler, Christopher C. (2012). Detection and identification of bacteria in clinical samples by
16S rRNA gene sequencing: comparison of two different approaches in clinical practice. Journal of Medical Microbiology
61(4). https://doi.org/10.1099/jmm.0.030387-0
31. Jiang, Shiman; Cai, Lingzhi; Lv, Longxian, and Li, Lanjuan. (2021). Pediococcus pentosaceus, a future additive or
probiotic candidate. Microbial Cell Factories, 20:45, https://doi.org/10.1186/s12934-021-01537-y
32. Johnson, Jethro S.; Spakowicz, Daniel J.; Hong, Bo-Young; Petersen, Lauren M; Demkowicz, Patrick; Chen, Lei; Leopold,
Shana R; Hanson, Blake M.; Agresta, Hanako O; Gerstein, Mark; Sodergren, Erica; and Weinstock, George M. (2019).
Evaluation of 16S rRNA gene sequencing for species and strain-level microbiome analysis, Nature Communications,
10:5029 | https://doi.org/10.1038/s41467-019-13036-1 www.nature.com/naturecommunications
33. Jose, N. M., Bunt, C. R., and Hussain, M. A. (2015). Comparison of Microbiological and Probiotic Characteristics of
Lactobacilli Isolates from Dairy Food Products and Animal Rumen Contents. Microorganisms, 3(2), 198-212.
https://doi.org/10.3390/microorganisms3020198
34. Khushboo, Karnwal A and Malik, T. (2023). Characterization and selection of probiotic lactic acid bacteria from different
dietary sources for development of functional foods. Front. Microbiol. 14:1170725. doi: 10.3389/fmicb.2023.1170725
35. Kwon, H.C.; Bae, H.; Seo, H.G.; Han, S.G. (2019). Chia seed extract enhances physiochemical and antioxidant properties
of yogurt. J. Dairy Sci., 102, 4870–4876.
36. Leboffe, Michael J. and Burton E. Pierce. (2010). Microbiology Laboratory Theory & Application 3e. Morton Publishing
Company, 772 pp.
37. Li, Mei-Na; Han, Qiang; Wang, Nan; Wang, Ting; You, Xue-Ming; Zhang, Shuai; Zhang, Cui-Cui; Shi, Yong-Qiang;
Qiao, Pei-Zhuang; Man, Cheng-Lian; Feng, Teng; Li, Yue-Yue; Zhu, Zhuang; Quan, Ke-Ji; Xu, Teng-Lin; Zhang, George
Fei. (2024). 16S rRNA gene sequencing for bacterial identification and infectious disease diagnosis. Biochemical and
Biophysical Research Communications, Volume 739, 150974, ISSN 0006-291X.
https://doi.org/10.1016/j.bbrc.2024.150974. (https://www.sciencedirect.com/science/article/pii/S0006291X24015109
38. Liu, C., W.J. Xue, H. Ding, C. An, S.J. Ma, and Y. Liu. (2022). Probiotic potential of Lactobacillus strains isolated from
fermented vegetables in Shaanxi, China, Front. Microbiol. 12, 774903, https://doi.org/10.3389/fmicb.2021.774903.
39. Mateos-Aparicio, I., Redondo-Cuenca, A., Villanueva-Suárez, M. J., & Zapata-Revilla, M. (2010). Pea pod, broad bean
pod, and okara, potential sources of functional compounds. LWT–Food Science and Technology, 43(9), 1467–1470.
40. Mathur, H., Beresford, T.P., and Cotter, P.D. (2020). Health Benefits of Lactic Acid Bacteria (LAB) Fermentates.
Nutrients 12:1679.
41. Medic, J., Atkinson, C., & Hurburgh, C. R. (2014). Current knowledge in soybean composition. Journal of the American
Oil Chemists’ Society, 91(3), 363–384. https://doi.org/10.1007/s11746-013-2407-9
42. Melini, F., Melini, V., Luziatelli, F., Ficca, A.G., and Ruzzi, M. (2019). Health-promoting components in fermented foods:
an up-to-date systematic review. Nutrients 11(5):1189. https://doi.org/10.3390/nu11051189
43. Meanti, F.; Mussio, C.; Rocchetti, G.; Rebecchi, A.; Lucini, L.; Morelli, L. (2024). Oat Okara Fermentation: New Insights
into the Microbiological and Metabolomic Characterization. Fermentation 10:545. https://
doi.org/10.3390/fermentation10110545
44. Mgomi, Fedrick C; Yuan, Lei; Wang, Yang; Rao, Sheng-Qi; and Yang, Zhen-Quan. (2022). Physiological properties,
survivability, and genomic characteristics of Pediococcus pentosaceus for application as a starter culture. International
Journal of Dairy Technology 75 (3): 588-602. https://doi.org/10.1111/1471-0307.12864
45. Montemurro, M., Celano, G., De Angelis, M., Gobbetti, M., Rizzello, C. G., and Pontonio, E. (2020). Selection of non-
lactobacillus strains to be used as starters for sourdough fermentation. Food Microbiol. 90:103491.
doi:10.1016/j.fm.2020.103491
46. Ngouenam, R.J., Nofal, G., Patra, S., Bilkissou, N., Marius, F., Marie, K., and Francois, Zambou. (2024). Characterization
of Lactic Acid Bacteria Isolated From Rotting Oranges and Use of Agropastoral Processing By-products as Carbon and
Nitrogen Sources Alternative for Lactic Acid Production. BioMed Research International. 2024. 10.1155/2024/4264229.
47. Ngouénam, Romial Joel; Nofal, Ghadir; Patra, Sanjukta; Njapndounke, Bilkissou; Kouam, Edith Marius Foko; Kaktcham,
Pierre Marie; and Ngoufack, François Zambou. (2024). Characterization of Lactic Acid Bacteria Isolated From Rotting
INTERNATIONAL JOURNAL OF LATEST TECHNOLOGY IN ENGINEERING,
MANAGEMENT & APPLIED SCIENCE (IJLTEMAS)
ISSN 2278-2540 | DOI: 10.51583/IJLTEMAS | Volume XIV, Issue VII, July 2025
www.ijltemas.in Page 907
Oranges and Use of Agropastoral Processing By-products as Carbon and Nitrogen Sources Alternative for Lactic Acid
Production. Wiley BioMed Research International, Article ID 4264229, 16 pages. https://doi.org/10.1155/2024/4264229
48. Olusola, Abiona Stella and Adepoju, Babarinde Yinusa. (2022). The Use of Lactic Acid Bacteria (LAB) Isolates as Single
and Mixed-Strain Starter Culture in Yoghurt Processing. Food Proc Nutr Sci 3(1): 15-21.
49. Othman, M., Ariff, A. B., Rios-Solis, L., & Halim, M. (2017). Extractive fermentation of lactic acid in lactic acid bacteria
cultivation: A review. Frontiers in Microbiology, 8, 2285.
50. Orf, J. (2010). Introduction. In Bilyeu, K., Ratnaparkhe, M. B., & Kole, C. (Eds.), Genetics, genomics, and breeding of
soybean. CRC Press.
51. Plessas, S., Mantzourani, I., and Bekatorou, A. (2020). Evaluation of Pediococcus pentosaceus SP2 as Starter Culture on
Sourdough Bread Making. Foods 9. doi: 10.3390/foods9010077
52. Prastujati, A. U., Hilmi, M., Khusna, A., Arief, I. I., Makmur, S., and Maulida, Q. (2022). Isolation and identification of
lactic acid bacteria of Bekamal (Banyuwangi traditional fermented meat). IOP Conference Series: Earth and
Environmental Science, 1020, 012026. https://doi.org/10.1088/1755-1315/1020/1/012026.
53. Qi Y, Huang L, Zeng Y, Li W, Zhou D, Xie J, Xie J, Tu Q, Deng D and Yin J. (2021). Pediococcus pentosaceus: Screening
and Application as Probiotics in Food Processing. Front. Microbiol. 12:762467. doi: 10.3389/fmicb.2021.762467
54. Ramadhanti, Nurazizah; Melia, Sri; Hellyward, James; and Purwati, Endang. (2021). Characteristics of lactic acid bacteria
isolated from palm sugar from West Sumatra, Indonesia and their potential as a probiotic. Biodiversitas, 22(5): 2610-2616.
ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d220520
55. Retnowati, Faizah Diah; Purwestri, Yekti Asih; and Sine, Yuni. (2024). Characterization of Lactic Acid Bacteria Isolated
from Soymilk and Its Growth in Soymilk By-product Medium for the Application in Soymilk Fermentation. Journal of
Tropical Biodiversity and Biotechnology, Volume 09, Issue 04: jtbb89003 DOI: 10.22146/jtbb.89003
56. Rezac, S., Kok, C.R., Heermann, M., and Hutkins, R. (2018). Fermented foods as a dietary source of live organisms. Front
Microbiol 9: 1785, DOI: https://doi.org/10.3389/fmicb.2018.01785.
57. Risna, Yayuk Kurnia; Harimurti, Sri; Wihandoyo; and Widodo. (2020). Screening for probiotic of lactic acid bacteria
isolated from the digestive tract of a native Aceh duck (Anas platyrhynchos). Biodiversitas, 21(7) : 3001-3007. ISSN:
1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d210717
58. Roslan, I.N.D.; Kamaruding, N.A.; Ismail, N.; and Shaharuddin, S. (2020). Sensory Attributes and Other Properties of
Yogurt Fortified with Immobilized Lactobacillus Plantarum and Soybean Residue (Okara). International Journal of
Probiotics and Prebiotics, Vol. 16, pp. 1–6, 2021, doi: https://doi.org/10.37290/ijpp2641-7197.16:1–6, ISSN 1555-1431
print; ISSN 2641-7197 online, Copyright © 2021 by New Century Health Publishers, LLC,
www.newcenturyhealthpublishers.com
59. Satari, B.; Karimi, K. (2018). Citrus processing wastes: Environmental impacts, recent advances, and future perspectives
in total valorization. Resour. Conser.Recycl., 129, 153–167.
60. Schleifer, K. H., & Ludwig, W. (1995). Phylogeny of the genus Lactobacillus and related genera. Systematic and Applied
Microbiology, 18(4): 461-467.
61. Sharma, R., P. Garg, P. Kumar, S.K. Bhatia, and S. Kulshrestha. (2020). Microbial Fermentation and Its Role in Quality
Improvement of Fermented Foods. Fermentation 2020, 6(4), 106; https://doi.org/10.3390/fermentation6040106
62. Septembre-Malaterre, A., Remize, F., & Poucheret, P. (2018). Fruits and vegetables as a source of nutritional compounds
and phytochemicals: Changes in bioactive compounds during lactic fermentation. Food Research International, 104, 86–
99.
63. Sevilla, John Russel G.; Esteban, Michael Angelo S.; Iñigo, Honey Bhabes R.; Orillaza, Audrey Mae V.; and Navarro,
Baby Richard R. (2021). 16S rRNA Gene Sequence Analysis of Acetic and Lactic Acid Bacteria Isolated from Philippine
Sugarcane Wine (Basi). PHILIPP AGRIC SCIENTIST, 104 (1): 75-81. ISSN 0031-7454
64. Tiensin, H., Kalibata, A., & Cole, M. (2020, 08/04/2020). Ensuring Food Security in the Era of COVID-19. Retrieved
from https://www.un.org/sustainabledevelopment/blog/2020/04/ensuring-foodsecurity-covid-19/
65. Ugwona, F.U, Obeta, N.A, Ejinkeonye, U.B and Aliyu, S.A. (2018). Production and Quality Evaluation of Soymilk
Yoghurt. Nigerian Journal of Nutritional Sciences, 39 (1).
66. Utami, Tyas; Cindarbhumi, Amaralda; Khuangga, Marcella C; Rahayu, Endang S; Cahyanto, Muhammad Nur; Nurfiyani,
Sri; and Zulaichah, Eni. (2020). Preparation of Indigenous Lactic Acid Bacteria Starter Cultures for Large Scale Production
of Fermented Milk. Digital Press Life Sciences 2: 00010. 10th Asian Conference of Lactic Acid Bacteria.
https://doi.org/10.29037/digitalpress.22327
67. Valin, H., Sands, R. D., van der Mensbrugghe, D., Nelson, G. C., Ahammad, H., Blanc, E., . . . Willenbockel, D. (2014).
The future of food demand: understanding differences in global economic models. Agricultural Economics, 45(1), 51-67.
doi:https://doi.org/10.1111/agec.12089
68. Venkateshwari, S., Halami, P. M., and Vijayendra, S. V. (2010). Characterisation of the heat-stable bacteriocin-producing
and vancomycin-sensitive Pediococcus pentosaceus CFR B19 isolated from beans. Benef. Microbes 1, 159–164. doi:
10.3920/bm2009.0032
69. Wahyuni, S., Ratna, Holilah, Asranudin, and Raharjo, Raden Alip. 2016. Antioxidant Activity of Isoflavones from Tofu
Pulp Waste. 6th Int'l Conference on Agriculture, Environment and Biological Sciences (ICAEBS'16) Dec. 21-22, 2016
Kuala Lumpur (Malaysia)
INTERNATIONAL JOURNAL OF LATEST TECHNOLOGY IN ENGINEERING,
MANAGEMENT & APPLIED SCIENCE (IJLTEMAS)
ISSN 2278-2540 | DOI: 10.51583/IJLTEMAS | Volume XIV, Issue VII, July 2025
www.ijltemas.in Page 908
70. Xu, X., Bi, S., Lao, F., Chen, F., Liao, X., and Wu, J. (2021). Comprehensive investigation on volatile and non-volatile
metabolites in broccoli juices fermented by animal- and plant-derived Pediococcus pentosaceus. Food Chem. 341:128118.
doi: 10.1016/j.foodchem.2020.128118
71. Xu, X. L. (2020). Research on key aroma identification and regulation of traditional sour curd cheese milk fan. Master’s
thesis. Shanghai University of Applied Technology, Shanghai, China.
72. Yamamoto, N., Shoji, M., Hoshigami, H., Watanabe, K., Watanabe, K., Takatsuzu, T., et al. (2019). Antioxidant capacity
of soymilk yogurt and exopolysaccharides produced by lactic acid bacteria. Biosci. Microbiota. Food Health 38, 97–104.
doi: 10.12938/bmfh.18-017
73. Yang, Yang; Xia, Yanan; Wang, Yu Rong; Sun, Li Shan; Shuang, Quan; and Zhang, Feng Mei. (2024). Optimization of
lactic acid bacterial starter culture to improve the quality and flavor characteristics of traditional Hurood. J. Dairy Sci.
107:105–122. https://doi.org/10.3168/jds.2023-23754
74. Zhou, R.Y., X. Huang, Z. Liu, J-Y Chua, S-Q Liu. (2022). Evaluating the effect of lactic acid bacterial fermentation on
salted soy whey for development of a potential novel soy sauce-like condiment. Current Research in Food Science, 5:1826-
1836. Elsevier