Selection of Proteolytic Lactic Acid Bacteria and the Potential as α-Glucosidase Inhibitor Activity During Milk Fermentation

https://doi.org/10.22146/agritech.100617

Miftakhussolikhah Miftakhussolikhah(1), Tyas Utami(2), Puspita Lisdiyanti(3), Endang Sutriwati Rahayu(4*)

(1) Faculty of Agricultural Technology, Universitas Gadjah Mada, Jl. Flora No. 1, Bulaksumur, Yogyakarta 55281; Research Center for Food Technology and Processing (PRTPP), National Research and Innovation Agency (BRIN), Jl. Jogja-Wonosari Km. 31,5 Gading, Playen, Gunungkidul, Yogyakarta 55861
(2) Faculty of Agricultural Technology, Universitas Gadjah Mada, Jl. Flora No. 1, Bulaksumur, Yogyakarta 55281
(3) Research Center for Biosystematics and Evolution, National Research and Innovation Agency (BRIN), Jl. Raya Jakarta-Bogor Km. 46 Cibinong 16911
(4) Faculty of Agricultural Technology, Universitas Gadjah Mada, Jl. Flora No. 1, Bulaksumur, Yogyakarta 55281
(*) Corresponding Author

Abstract


Diabetes mellitus (DM) is a degenerative condition increasing globally, including in Indonesia, with a corresponding rise in death rates. One of the therapy methods for DM is the use of α-glucosidase inhibitor (AGI), with peptides identified as potential AGI-based food ingredients. Therefore, this study aimed to develop fermented milk products enhanced with AGI peptides by screening and identifying lactic acid bacteria (LAB) strains with proteolytic capabilities. The experiment was conducted in three main stages: (1) screening 32 LAB isolates for their proteolytic activity, (2) determining the optimal fermentation time for AGI production using selected proteolytic LAB strains, and (3) molecular identification of the selected LAB strains through 16S rRNA gene sequencing. The results showed that two isolates, SR17B and L23, identified as Lactiplantibacillus plantarum-pentosus SR17B and Lactiplantibacillus plantarum-pentosus L23, had high proteolytic activity and were capable of producing fermented milk with significant AGI activity of 35.94% and 35.15%, respectively. AGI activity progressively intensified during fermentation, peaking at 12 hours in both strains, indicating that this period of time was ideal for fermentation in order to have the largest inhibitory effect. These results suggested that selected LAB strains, particularly L. plantarum-pentosus SR17B and L23, could serve as functional starter cultures for development of fermented dairy products with antidiabetic potential.


Keywords


α-glucosidase inhibitor; fermentation; lactic acid bacteria; milk; proteolytic

Full Text:

PDF


References

Aliifah, F., Rustama, M. M. & Setiyadi. W. P. (2023). Skrining bakteri asam laktat dan yeast potensial proteolitik ekstraseluler dan milk clotting activity dari getah pepaya (Carica papaya L.) dan fresh cheese. Jurnal Teknologi Hasil Peternakan, 4(2), 167-186. https://doi.org/10.24198/jthp.v4i2.49944

Atanasova, J., Moncheva, P. & Ivanova, I. (2014). Proteolytic and antimicrobial activity of lactic acid bacteria grown in goat milk. Biotechnology and Biotechnological Equipment, 28, 1073-1078. https://doi.org/10.1080/13102818.2014.971487

Baba, W.N., Mudgil, P., Kamal, H., Kilari, B. P., Gan, C. Y. & Maqsood, S. (2021). Identification and characterization of novel α-amylase and α-glucosidase inhibitory peptides from camel whey proteins. Journal of Dairy Science, 104(2), 1364-1377. https://doi.org/10.3168/jds.2020-19271

Behbahani, B. A., Noshad, M., Namazi, P. & Vasiee, A. (2024). Exploring the probiotic potential of Lactiplantibacillus pentosus SM1: Resistance, anti-microbial activity, anti-biofilm, cytotoxic activity, and safety properties. Food Science and Technology, 210, 116850. https://doi.org/10.1016/j.lwt.2024.116850

Botha, S.P. & Bigwood, E.J. (1959). Amino-acid content of raw and heat-sterilized cow’s milk. British Journal of Nutrition, 13, 385-389. https://doi.org/10.1079/BJN19590052

Cao, R., Li, W., Zhang, J., Bao, X., Feng, H., Sun, J., Liu, X. & Sun, L. (2024). Milk casein hydrolysate peptides regulate starch digestion through inhibition of α-glucosidase: An insight into the active oligopeptide screening, enzyme inhibition behaviors, and oligopeptide-enzyme binding interactions. Food Hydrocolloids, 152. https://doi.org/10.1016/j.foodhyd.2024.109926

Chen, P., Zhang, Q., Dang, H., Liu, X., Tian, F., Zhao, J., Chen, Y., Zhang, H. & Chen, W. 2014. Screening for potential new probiotic based on probiotic properties and a-glucosidase inhibitory activity. Food Control, 35, 65-72. https://doi.org/10.1016/j.foodcont.2013.06.027

Dean, W. M. D. (2017). Acarbose: Anti-diabetic, Cardio-protective, Weight loss, and Potential Anti-aging agent. Magazine life enhancement. https://www.warddeanmd.com/acarbose-antidiabetic-cardioprotective-weight-loss-potential-antiaging-agent/ (accessed on 18 February 2024)

De Valdez, G.F., De Giori, G.S., Holgado, A.P.D.R. & Oliver, G. (1985). Effect of the Rehydration Medium on the Recovery of Freeze-Dried Lactic Acid Bacteria. Applied and Environmental Microbiology, 50(5), 1339-1341. https://doi.org/10.1128%2Faem.50.5.1339-1341.1985

Echegaray, N., Yilmaz, B., Sharma, H., Kumar, M., Pateiro, M., Ozogul, F. & Lorenzo, J.M. (2023). A novel approach to Lactiplantibacillus plantarum: from probiotic properties to the omics insights. Microbiological Research, 268, 127289. https://doi.org/10.1016/j.micres.2022.127289

George, F., Daniel, C., Thomas, M., Singer, E., Guilbaud, A., Tessier, F.J., Juneless, A.M.R., Borges, F. & Foligne, B. (2018). Occurrence and dynamism of lactic acid bacteria in distinct ecological niches: a multifaceted functional health perspective. Frontiers in Microbiology, 9, 2899. https://doi.org/10.3389/fmicb.2018.02899

Guo, W., Xiao, Y., Fua, X., Long, Z., Wua, Y., Lin, Q., Rena, K. & Jiang, L. (2023). Identification of novel α-glucosidase and ACE inhibitory peptides from Douchi using peptidomics approach and molecular docking. Food Chemistry, 19, 100779. https://doi.org/10.1016/j.fochx.2023.100779

Ha, T. J., Park, J. E., Kang, B. K., Kim, H. S., Shin, S. O., Seo, J. H., Oh, E., Kim, S. & Kwak, D. (2019). α-Glucosidase Inhibitory Activity of Isoflavones and Saponins from Soybean (Glycine max L.) and Comparisons of Their Constituents during Heat Treatments. Journal of the Korean Society of Food Science and Nutrition, 48(9), 953-960. https://doi.org/10.3746/jkfn.2019.48.9.953

Hengkengbala, S. I., Lintang, R. A. J., Sumilat, D. A., Mangindaan, R. E. P., Ginting, E. L. & Tumembouw, S. (2021). Karakteristik morfologi dan aktivitas enzim protease bakteri simbion nudibranch. Jurnal Pesisir dan Laut Tropis, 9 (3). https://doi.org/10.35800/jplt.9.3.2021.36672

Horie, M., Sato, H., Tada, A., Nakamura, S., Sugino, S., Tabei, Y., Katoh, M. & Toyotome, T. (2019). Regional characteristics of Lactobacillus plantarum group strains isolated from two kinds of Japanese post-fermented teas, Ishizuchi-kurocha and Awa-bancha. Bioscience of Microbiota, Food and Health Vol. 38 (1), 11–22. https://doi.org/10.12938%2Fbmfh.18-005

Ibrahim, M. A., Bester, M. J., Neitz, A. W. H. & Gaspar, A. R. M. (2017). Structural properties of bioactive peptides with α-glucosidase inhibitory activity. Chemical Biology and Drug Design, 91(2), 370-379. https://doi.org/10.1111/cbdd.13105

IDF (International Diabetes Federation). (2022). Diabetes around the world in 2021. https://diabetesatlas.org (accessed on 20 July 2024)

Karyantina, M., Anggrahini, S., Utami, T. & Rahayu, E.S. (2020) Moderate halophilic lactic acid bacteria from jambalroti: a traditional fermented fish of Central Java, Indonesia. Journal of Aquatic Food Product Technology. https://doi.org/10.1080/10498850.2020.1827112

Kieliszek, M., Kolotylo, V., Szczyrba, A.M., Giurgiulescu, L., Kot, A.M., Kalisz, S., Pobiega, K. & Cendrowski, A. (2021). Isolation and identification of new yeast strains from bee bread. Carpathian Journal of Food Science and Technology. https://doi.org/10.34302/crpjfst/2021.13.1.17

Kirilov, N., Petkova, T., Atanasova, J., Danova, S.V., Iliev, I., Popov, Y., Haertle, Y. & Ivanova, I.V. (2009). Proteolytic activity in lactic acid bacteria from Iraq, Armenia and Bulgaria. Biotechnology and Biotechnological Equipment, 23, 643-646. https://doi.org/10.1080/13102818.2009.10818506

Konrad, B., Anna, D. B., Marek, S., Marta, P., Aleksandra, Z. & Jo´zefa, C. (2014). The evaluation of dipeptidyl peptidase (DPP)-IV, α-glucosidase and angiotensin converting enzyme (ACE) inhibitory activities of whey proteins hydrolyzed with serine protease isolated from Asian pumpkin (Cucurbita ficifolia). International Journal of Peptide Research and Therapeutic, 20, 483-491. https://doi.org/10.1007/s10989-014-9413-0

Li, W., Fu, X., Zhang, T., Li, H., Chen, T. & Liu, X. (2023). Isolation and identification of an α-glucosidase inhibitory peptide from extruded soybean protein and its hypoglycemic activity in T2DM mice. Food and Function, 14 (9), 4288-4301. https://doi.org/10.1039/d3fo00580a

Liu, W., Li, H., Wen, Y., Liu, Y., Wang, J. & Sun, B. (2021). Molecular mechanism for the α‑glucosidase inhibitory effect of wheat germ peptides. Journal of Agricultural and Food Chemistry, 69, 15231−15239. https://doi.org/10.1021/acs.jafc.1c06098

Miftakhussolikhah, M., Utami, T., Lisdyanti, P., & Rahayu, E. S. (2025). Stability α-glucosidase inhibitory activity of fermented milk using Lactoplantibacillus strain L23 and SR17B during simulated gastrointestinal digestion and cold storage. BIO Web of Conferences, 177, 07004. https://doi.org/10.1051/bioconf/202517707004

Ministry of Health. (2018). Laporan Nasional Riskesdas 2018. https://layanandata.kemkes.go.id/katalog-data/riskesdas/ketersediaan-data/riskesdas-2018 (accessed on 20 Juli 2024)

Ministry of Health. (2023). Survei Kesehatan Indonesia Tahun 2023: dalam angka. https://layanandata.kemkes.go.id/katalog-data/ski/ketersediaan-data/ski-2023 (accessed on 20 Juli 2024)

NCBI. (2024). National Center for Biotechnology Information https://www.ncbi.nlm.nih.gov/. (accessed on 01 November 2024)

Ngamsomchat, A., Kaewkod, T., Konkit, M., Tragoolpua, Y., Bovonsombut, S. & Chitov, T. (2022). Characterisation of Lactobacillus plantarum of dairy-product origin for probiotic chèvre cheese production. Foods, 11, 934. https://doi.org/10.3390/foods11070934

Matti, A., Utami, T., Hidayat, C. & Rahayu, E. S. (2019). Isolation, screening and identification of proteolytic lactic acid bacteria from indigenous chao product. J Aquat Food Prod Technol. https://doi.org/10.1080/10498850.2019.1639872

Meneses, M. J., Silva, B. M., Sousa, M., Sá, R., Oliveira, P.F. & Alves, M. G. (2015). Antidiabetic Drugs: Mechanisms of Action and Potential Outcomes on Cellular Metabolism. Current Pharmaceutical Design, 21, 3606-3620. https://doi.org/10.2174/1381612821666150710145753

Mudgil, P., Kamal, H., Kilari, B.P., Salim, M.A.S.M., Gan, C.Y., Maqsood, S. (2021). Simulated gastrointestinal digestion of camel and bovine casein hydrolysates: Identification and characterization of novel anti-diabetic bioactive peptides. Food Chem, 353, 129374

Muganga, L., Liu, X., Tian, F., Zhao, J., Zhang, H. & Chen, W. (2015). Screening for lactic acid bacteria based on antihyperglycaemic and probiotic potential and application in synbiotic set yoghurt. Journal of Functional Foods, 16, 125–136. https://doi.org/10.1016/j.jff.2015.04.030

Rahmani, N., Sari, Y. N., Nurheni Sri Palupi2 & Yopi. (2013). Isolats Bakteri Indigenous Penghasil Milk-Clotting Protease untuk Fermentasi Keju (Isolates of Indigenous Bacteria Producing Milk-Clotting Protease for Cheese Fermentation). Jurnal Biologi Indonesia, 9(2), 199-208.

Rai, A. K., Sanjukta, S., Chourasia, R., Bhat, I., Bhardwaj, P.K. & Sahoo, D. (2017). Production of bioactive hydrolysate using protease, β-glucosidase and α-amylase of Bacillus spp. isolated from kinema. Bioresource Technology, 235, 358-365. https://doi.org/10.1016/j.biortech.2017.03.139

Ramchandran, L. & Shah, N. P. (2008). Proteolytic Profiles and Angiotensin-I Converting Enzyme and α-Glucosidase Inhibitory Activities of Selected Lactic Acid Bacteria. Journal of Food Science, 73 (2), 75-81. https://doi.org/10.1111/j.1750-3841.2007.00643.x

Ren, Y., Liang, K., Jin, Y., Zhang, M., Chen, Y., Wu, H. & Lai, F. (2016). Identification and characterization of two novel α-glucosidase inhibitory oligopeptides from hemp (Cannabis sativa L.) seed protein. Journal of Functional Foods, 26, 439-450. https://doi.org/10.1016/j.jff.2016.07.024

Sa’adah, A. & Muhtadi.(2022). Inhibitory Activity of the α-Glucosidase Enzyme by Albumin Isolated from Giant Gourami (Osphronemus Goramy), Rice Eel (Monopterus albus), and Mackerel Tuna (Euthynnus affinis). Proceedings of the 4th International Conference Current Breakthrough in Pharmacy (ICB-Pharma 2022). https://doi.org/10.2991/978-94-6463-050-3_15

Sabrini, Z., Rukmi, I., and Ferniah, R. S., 2021. Aktivitas Enzimatis Biakan Kapang Aspergillus Section Nigri DUCC (Diponegoro University Culture Collection) Dan Identifikasi Molekuler Isolat Potensial. Bioma: Berkala Ilmiah Biologi, [Online] Volume 23(1), pp. 1-5. https://doi.org/10.14710/bioma.23.1.1-5

Sapra, A. & Bhandari, P. 2023. Diabetes. https://www.ncbi.nlm.nih.gov/books/NBK551501/. Accessed on 20 August, 2024.

Savijoki, K. Ingmer, H. Varmanen, P. (2006). Proteolytic systems of lactic acid bacteria. Applied Microbiology and Biotechnology. 71: 394–406. https://doi.org/10.1007/s00253-006-0427-1

Sionek, B., Szydłowska, A., Trzaskowska, M. & Krajewska, D. K. (2024). Review: the impact of physicochemical conditions on lactic acid bacteria survival in food products fermentation, 10, 298. https://doi.org/10.3390/fermentation10060298

Spigaglia, P & Mastrantonio, P. (2003). Evaluation of Repetitive Element Sequence-Based PCR as a Molecular Typing Method for Clostridium difficile. Journal of Clinical Microbiology, 41(6), 2454–2457. doi: 10.1128/JCM.41.6.2454-2457.2003

Suhartatik, N., Cahyanto, M. N., Rahardjo, S., Miyashita, M. & Rahayu, E. S. (2014). Isolation and Identification of lactic acid bacteria producing β-glucosidase from indonesian fermented foods. International Food Research Journal, 21(3), 973–78. http://www.ifrj.upm.edu.my/21%20(03)%202014/19%20IFRJ%2021%20(03)%202014%20Suhartik%20376.pdf

Sun, H., Saeedi, P., Karuranga, S., Pinkepank, M., Ogurtsova, K., Duncan, B., Stein, C., Basit, A., Chan, J. C. N., Mbanya, J. C., Pavkov, M. E., Ramachandaran, A., Wild, S. H., James, S., Herman, W. H., Zhang, P., Bommer, C., Kuo, S., Boyko, E. J., & Magliano, D. J. (2022). IDF Diabetes Atlas: Global, regional and country-level diabetes prevalence estimates for 2021 and projections for 2045. Diabetes Research and Clinical Practice, 183. https://doi.org/10.1016/j.diabres.2021.109119

Vilcacundo, R., Villaluenga, C. M. & Ledesma, B.H. (2017). Release of dipeptidyl peptidase IV, α-amylase and α-glucosidase inhibitory peptides from quinoa (Chenopodium quinoa Willd.) during in vitro simulated gastrointestinal digestion. Journal of Functional Foods, 35, 531-539. https://doi.org/10.1016/j.jff.2017.06.024

Walter, H. E. (1984). Proteinases In Methods of enzymatic analysis, (ed. H.U. Bergmeyer and M Qrassl), Verlag Chemie. Weinheim-Deerfield, Florida-Basse, Vol. 5:271-276.

Wang, F., Zhang, Y., Yua, T., Hea, J., Cuia, J., Wanga, J., Cheng, X. & Fana, J. (2018). Oat globulin peptides regulate antidiabetic drug targets and glucose transporters in Caco-2 cells. Journal of Functional Foods, 42, 12-20. https://doi.org/10.1016/j.jff.2017.12.061

Wang, X., Dejun, Y., Zhang, Y., Zhang, C., Liu, L., Liu, Y., Jiang, J., Xie, P. & Huang L. (2023). Screening and Evaluation of Novel α‑Glucosidase Inhibitory Peptides from Ginkgo biloba Seed Cake Based on Molecular Docking Combined with Molecular Dynamics Simulation. Journal of Agricultural and Food Chemistry, 71, 10326−10337. https://doi.org/10.1021/acs.jafc.3c00826

Yin, Z., Zhang, W., Feng, F., Zhang, Y. & Kang, W. (2014). α-Glucosidase inhibitors isolated from medicinal plants. Food Science and Human Wellness, 3, 136–174. https://doi.org/10.1016/j.fshw.2014.11.003

Yuliana, T., Pratiwi, A.R., Zahratunnisa, S., Rialita, T., Cahyana, Y., Harlina, P.W. & Marta, H. (2023). Purification and partial characterization of a bacteriocin produced by Lactobacillus pentosus 124-2 Isolated from Dadih. Applied Sciences, 13, 4277. https://doi.org/10.3390/app13074277

Yusmarini, Indrati, R., Utami, T. & Marsono, Y. (2010). Aktivitas Proteolitik Bakteri Asam Laktat dalam Fermentasi Susu Kedelai. Jurnal Teknologi dan Industri Pangan, Vol XXI (2)

Yu, Z., Yin, Y., Zhao, W., Yu, Y., Liu, B., Liu, J. & Chen, F. (2011). Novel peptides derived from egg white protein inhibiting alpha-glucosidase. Food Chemistry, 129, 1376–1382. https://doi.org/10.1016/j.foodchem.2011.05.067

Yu, Z., Yin, Y., Zhao, W., Liu, J. & Chen, F. (2012). Anti-diabetic activity peptides from albumin against α-glucosidase and α-amylase. Food Chemistry, 135, 2078–2085. https://doi.org/10.1016/j.foodchem.2012.06.088

Zeng, Z., Luo, J., Zuo, F., Zhang, Y., Ma, H. & Chen, S. (2016). Screening for potential novel probiotic Lactobacillus strains based on high dipeptidyl peptidase IV and α-glucosidase inhibitory activity. Journal of Functional Foods, 20, 486–495. https://doi.org/10.1016/j.jff.2015.11.030

Zhang, Y., Wang, N., Wang, W., Wang, J., Zhua, Z. & Li, X. (2016). Molecular mechanisms of novel peptides from silkworm pupae that inhibit α-glucosidase. Peptides, 76, 45-50. https://doi.org/10.1016/j.peptides.2015.12.004

Ziaee, A., Esmailzadehha, N. & Honardoost, M. (2017). Comparison of adjunctive therapy with metformin and acarbose in patients with Type-1 diabetes mellitus. Pakistan Journal of Medical Sciences, 33(3), 686-690. https://doi.org/10.12669%2Fpjms.333.12669



DOI: https://doi.org/10.22146/agritech.100617

Article Metrics

Abstract views : 5 | views : 1

Refbacks

  • There are currently no refbacks.


Bookmark and Share


Copyright (c) 2026 Endang Sutriwati Rahayu

Creative Commons License
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

agriTECH has been Indexed by:

Emerging Source Citation Index Directory of Open Access Journals CABI.org Dimensions Google Scholar Sinta Garuda

agriTECH (print ISSN 0216-0455; online ISSN 2527-3825) is published by Faculty of Agricultural Technology, Universitas Gadjah Mada in colaboration with Indonesian Association of Food Technologies.

website statisticsView My Stats