Molecular Phylogenetic Analysis of 16S rRNA Gene of Facultative Hetero-Fermentative Lactic Acid Bacteria

Habib Ramadhan, Erna Wulandari

Abstract


Facultative heterofermentative lactic acid bacteria are bacteria that ferment carbohydrates to produce lactic acid and by-products such as acetic acid, ethanol, and CO2. This group of bacteria is widely used in the food and health industries. However, research on BAL species, especially the facultative heterofermentative group, is still relatively limited, so further studies are needed with molecular phylogenetic analysis aimed at providing an overview of the kinship of BAL species in the facultative heterofermentative group using the 16S rRNA gene molecular marker for more accurate identification results. The research design was conducted as a molecular-based descriptive analysis using secondary data in the form of 16S rRNA gene nucleotide sequences taken from the National Center for Biotechnology Information (NCBI) GenBank. Data analysis was performed using bioinformatics tools, including sequence alignment using ClustalX, phylogenetic tree reconstruction using MEGA and PHYLIP, and similarity matrix analysis using PHYDIT. The results of the analysis showed that Lactobacillus casei and Lactobacillus rhamnosus, as well as Lactobacillus pentosus and Lactobacillus plantarum, were closely related with a bootstrap value of 100% and a similarity matrix of 98-99%. Meanwhile, Lactobacillus alimentarius showed a relatively close relationship with Lactobacillus pentosus and Lactobacillus plantarum with bootstrap values and similarity matrices of around 92%. Thus, molecular phylogenetic analysis is effective in analyzing, identifying, and proving the relationship between BALs of the facultative heterofermentative group.


Keywords


Lactic acid bacteria; Facultative Hetero-Fermentative; Phylogenetic; Molecular; 16S Rrna Gene

Full Text:

PDF

References


Adamu, B. B., Odomu, G. I. B., Ideh, R. R., & Olukotun, G. B. (2025). The role of lactic acid bacteria in food, agriculture and industry: A review. GSC Biological and Pharmaceutical Sciences, 30(1), 099–106. https://doi.org/10.30574/gscbps.2025.30.1.0497

Ahmad, A., Banat, F., & Taher, H. (2020). A review on the lactic acid fermentation from low-cost renewable materials: Recent developments and challenges. In Environmental Technology and Innovation (Vol. 20). Elsevier B.V. https://doi.org/10.1016/j.eti.2020.101138

Akihary, C. V., & Kolondam, B. J. (2020). PEMANFAATAN GEN 16S rRNA SEBAGAI PERANGKAT IDENTIFIKASI BAKTERI UNTUK PENELITIAN-PENELITIAN DI INDONESIA. PHARMACON, 9(1). https://doi.org/10.35799/pha.9.2020.27405

Axelsson, L. (2004). Lactic acid bacteria: Classification and physiology. In Lactic Acid Bacteria Microbiological and Functional Aspects, Third Edition: Revised and Expanded (pp. 1–66). CRC Press.

Ayivi, R. D., Gyawali, R., Krastanov, A., Aljaloud, S. O., Worku, M., Tahergorabi, R., Silva, R. C. da, & Ibrahim, S. A. (2020). Lactic Acid Bacteria: Food Safety and Human Health Applications. Dairy, 1(3), 202–232. https://doi.org/10.3390/dairy1030015

Baldwin, B. G., Sanderson, M. J., Porter, J. M., Wojciechowski, M. F., Campbell, C. S., & Donoghue, M. J. (1995). The its Region of Nuclear Ribosomal DNA: A Valuable Source of Evidence on Angiosperm Phylogeny. Annals of the Missouri Botanical Garden, 82(2), 247. https://doi.org/10.2307/2399880

Bernardeau, M., Guguen, M., & Vernoux, J. P. (2006). Beneficial lactobacilli in food and feed: Long-term use, biodiversity and proposals for specific and realistic safety assessments. In FEMS Microbiology Reviews (Vol. 30, Issue 4). https://doi.org/10.1111/j.1574-6976.2006.00020.x

Bintsis T. (2018). Lactic acid bacteria as starter cultures: An update in their metabolism and genetics. AIMS microbiology, 4(4), 665–684. https://doi.org/10.3934/microbiol.2018.4.665.

Bramasta, R. C., Faiqoh, E., Hendrawan, I. G., Sembiring, A., & Yusmalinda, N. L. A. (2021). Identifikasi Hiu yang Diperdagangkan di Bali Menggunakan Metode DNA Barcoding dan Analisis Filogenetik. Journal of Marine and Aquatic Sciences, 7(1), 84. https://doi.org/10.24843/jmas.2021.v07.i01.p12

Chase, M. W., Soltis, D. E., Olmstead, R. G., Morgan, D., Les, D. H., Mishler, B. D., Duvall, M. R., Price, R. A., Hills, H. G., Qiu, Y.-L., Kron, K. A., Rettig, J. H., Conti, E., Palmer, J. D., Manhart, J. R., Sytsma, K. J., Michaels, H. J., Kress, W. J., Karol, K. G., … Albert, V. A. (1993). Phylogenetics of Seed Plants: An Analysis of Nucleotide Sequences from the Plastid Gene rbcL. Annals of the Missouri Botanical Garden, 80(3), 528. https://doi.org/10.2307/2399846

Chun, J. (1995). Computer-assisted classification and identification of actinomycetes. Newcastle upon Tyne, UK: University of Newcastle. Ph. D. thesis.

Dykhuizen, D. (2005). Species Numbers in Bacteria. Proceedings. California Academy of Sciences, 56(6 Suppl 1), 62–71. Https://Pmc.Ncbi.Nlm.Nih.Gov/Articles/PMC3160642/#S3.

Felsenstein, J. (2005). J. Felsenstein, Inferring Phylogenies, Sinauer Assoc., 2004, pp. xx + 664. Journal of Classification, 22(1), 139–142. https://doi.org/10.1007/s00357-005-0009-4

Finanda, A., Mukarlina, & Rahmawati. (2021). ISOLASI DAN KARAKTERISASI GENUS BAKTERI ASAMLAKTAT DARI FERMENTASI DAGING BUAH PISANG KEPOK (Musa paradisiaca L.). Jurnal Protobiont, 10(2), 37–41.

Hidayat, T., & Pancoro, A. "ULASAN Kajian Filogenetika Molekuler Dan Peranannya Dalam Menyediakan Informasi Dasar Untuk Meningkatkan Kualitas Sumber Genetik Anggrek." Jurnal AgroBiogen, vol. 4, no. 1, Apr. 2008, pp. 35-40, doi:10.21082/jbio.v4n1.2008.p35-40.

Hill, D., Sugrue, I., Tobin, C., Hill, C., Stanton, C., & Ross, R. P. (2018). The Lactobacillus casei Group: History and Health Related Applications. Frontiers in Microbiology, 9. https://doi.org/10.3389/fmicb.2018.02107

Hillis, D. M., & Bull, J. J. (1993). An Empirical Test of Bootstrapping as a Method for Assessing Confidence in Phylogenetic Analysis. Systematic Biology, 42(2), 182–192. https://doi.org/10.2307/2992540

Huang, C.-H., Li, S.-W., Huang, L., & Watanabe, K. (2018). Identification and Classification for the Lactobacillus casei Group. Frontiers in Microbiology, 9. https://doi.org/10.3389/fmicb.2018.01974

Jumrah, E., & Salnus, S. (2024). (Review Article) Identification of Endophytic Bacteria by 16S rRNA Analysis. Jurnal Sains Dan Teknik Terapan, 2(2).

Karyawati, A. T., Darmakusuma, D., Mauboy, R. S., Amalo, D., & Mutis, A. (2024). Potensi Bakteri Asam Laktat sebagai Penghasil Senyawa Antibakteri dan Enzim Ekstraseluler. Jurnal Biotropikal Sains, 21(2).

Kusnadie, B. M., Purwanti, N. U., & Liana, D. F. (2025). ISOLASI DAN KARAKTERISASI BAKTERI ASAM LAKTAT PADA MAKANAN FERMENTASI PEKASAM IKAN SELUANG (Rasbora sp.). Jurnal Sains Dan Teknologi Pangan, 10(2). https://doi.org/10.63071/fzxbv993

Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., Mcgettigan, P. A., McWilliam, H., Valentin, F., Wallace, I. M., Wilm, A., Lopez, R., Thompson, J. D., Gibson, T. J., & Higgins, D. G. (2007). Clustal W and Clustal X version 2.0. Bioinformatics, 23(21). https://doi.org/10.1093/bioinformatics/btm404

Meurant, G. (2011). Computer-Assisted Bacterial Systematics. Los Angeles; Academy Press.

Mokoena, M. P. (2017). Lactic Acid Bacteria and Their Bacteriocins: Classification, Biosynthesis and Applications against Uropathogens: A Mini-Review. Molecules, 22(8), 1255. https://doi.org/10.3390/molecules22081255

Moritz, C. & Hillis, D. M. (1996). Molecular systematics: Context and controversies. (2nd edition). Sinauer Associate.

Muthi’ah, S. N., & Jannah, M. (2023). ANALISIS FILOGENETIK PADA SPESIES JERUK (Citrus sp.) BERDASARKAN SEKUENS ITS SECARA IN SILICO. BIO-SAINS : Jurnal Ilmiah Biologi, 2(2), 62–66. https://doi.org/10.34005/bio-sains.v2i2.2106

Noer, S. (2021). Identifikasi Bakteri secara Molekular Menggunakan 16S rRNA. EduBiologia: Biological Science and Education Journal, 1(1). https://doi.org/10.30998/edubiologia.v1i1.8596

Nurkanto, A., & Agusta, A. (2015). Identifikasi Molekular dan Karakterisasi Morfo-Fisiologi Actinomycetes Penghasil Senyawa Antimikroba. Jurnal Biologi Indonesia, 11(2), 195–203. http://www.ncbi.nlm.nih.gov,

Prakash, O., Jangid, K., & Shouche, Y. S. (2013). Carl Woese: from Biophysics to Evolutionary Microbiology. Indian Journal of Microbiology, 53(3), 247–252. https://doi.org/10.1007/s12088-013-0401-4

Quinto, E. J., Jiménez, P., Caro, I., Tejero, J., Mateo, J., & Girbés, T. (2014). Probiotic Lactic Acid Bacteria: A Review. Food and Nutrition Sciences, 05(18), 1765–1775. https://doi.org/10.4236/fns.2014.518190

Sahadeva, M. L., & Pertiwi, N. P. D. (2024). Konstruksi Pohon Filogenetik Spesies dalam Famili Orchidaceae Berdasarkan Marka Gen matK Kloroplas: Studi in Silico. Wahana Matematika Dan Sains: Jurnal Matematika, Sains, Dan Pembelajarannya, 17(3). https://doi.org/10.23887/wms.v17i3.87986

Subari, A., Razak, A., & Sumarmin, R. (2021). Phylogenetic Analysis of Rasbora spp. Based on the Mitochondrial DNA COI gene in Harapan Forest. Jurnal Biologi Tropis, 21(1), 89–94. https://doi.org/10.29303/jbt.v21i1.2351

Surak, A. F., Ndaong, N. A., & Detha, A. I. R. (2024). Studi Literatur Bakteri Asam Laktat Yang Diisolasi Dari Susu Kuda, Susu Kambing Dan Susu Sapi. Jurnal Veteriner Nusantara, 7(30), 1–10.

Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., & Kumar, S. (2011). MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28(10), 2731–2739. https://doi.org/10.1093/molbev/msr121

Tindi, M., Mamangkey, N. G. F., & Wullur, S. (2017). DNA Barcode dan analisis filogenetik molekuler beberapa jenis bivalvia asal perairan Sulawesi Utara berdasarkan gen COI. JURNAL PESISIR DAN LAUT TROPIS, 5(2). https://doi.org/10.35800/jplt.5.2.2017.15050

Ubaidillah, R. dan Sutrisno H. 2009. Pengantar Biosistemik: Teori dan Praktikum. LIPI Press, Jakarta.

Woese, C. R., Stackebrandt, E., Macke, T. J., & Fox, G. E. (1985). A Phylogenetic Definition of the Major Eubacterial Taxa. Systematic and Applied Microbiology, 6(2). https://doi.org/10.1016/S0723-2020(85)80047-3

Yang, Z., & Rannala, B. (2012). Molecular phylogenetics: principles and practice. Nature Reviews Genetics, 13(5), 303–314. https://doi.org/10.1038/nrg3186

Zou, Y., Zhang, Z., Zeng, Y., Hu, H., Hao, Y., Huang, S., & Li, B. (2024). Common Methods for Phylogenetic Tree Construction and Their Implementation in R. In Bioengineering (Vol. 11, Issue 5). https://doi.org/10.3390/bioengineering11050480




DOI: https://doi.org/10.14421/biomedich.2026.151.1641-1647

Refbacks

  • There are currently no refbacks.




Copyright (c) 2026 Habib Ramadhan, Erna Wulandari



Biology, Medicine, & Natural Product Chemistry
ISSN 2089-6514 (paper) - ISSN 2540-9328 (online)
Published by Sunan Kalijaga State Islamic University & Society for Indonesian Biodiversity.

CC BY NC
This work is licensed under a CC BY-NC