Identifikasi kondisi induksi plantaricin dari lactiplantibacillus plantarum su-kc1a dengan metode co-culture = Identification of plantaricin induced conditions from lactiplantibacillus plantarum su-kc1a by co-culture method

Candra, Tasya Mariska (2024) Identifikasi kondisi induksi plantaricin dari lactiplantibacillus plantarum su-kc1a dengan metode co-culture = Identification of plantaricin induced conditions from lactiplantibacillus plantarum su-kc1a by co-culture method. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Bakteriosin adalah peptida antimikroba yang diproduksi oleh bakteri asam laktat dan memiliki potensi sebagai pengganti antibiotik serta pengawet makanan kimiawi. Salah satu jenis bakteriosin kelas II yaitu plantaricin dihasilkan oleh Lactiplantibacillus plantarum. Beberapa gen pengode plantaricin telah diidentifikasi pada genom L. plantarum SU-KC1a, namun faktor yang dapat menginduksi ekspresi plantaricin belum diketahui. Salah satu metode untuk meningkatkan produksi plantaricin adalah melalui metode co-culture. Penelitian ini bertujuan untuk mengidentifikasi kondisi optimal untuk induksi plantaricin dari L. plantarum SU-KC1a melalui co-culture dengan tiga bakteri asam laktat, yaitu Enterococcus faecalis A23, Lactobacillus fermentum E5, dan Leuconostoc mesenteroides S2. Pada penelitian ini juga akan ditentukan waktu optimum untuk pertumbuhan serta produksi plantaricin menggunakan uji aktivitas antibakteri dari Cell-Free Supernatant (CFS) dengan metode difusi sumuran dan broth dilution. Selain itu, akan dilakukan presipitasi protein dengan menggunakan ammonium sulfat dan dilanjutkan dengan identifikasi berat molekul plantaricin dengan Tricine SDS-PAGE. Hasil penelitian menunjukkan bahwa aktivitas antibakteri yang terdeteksi dengan metode difusi sumuran sebagian besar disebabkan oleh asam organik. Akan tetapi, setelah presipitasi protein dan uji antibakteri dengan metode broth dilution, ditemukan adanya aktivitas antibakteri yang diduga berasal dari peptida, kemungkinan plantaricin dari L. plantarum SU-KC1a. Pemisahan protein dengan Tricine SDS-PAGE juga menghasilkan pita protein dengan berat molekul sekitar 5 kDa, yang diduga sebagai plantaricin. Selain itu, perbandingan aktivitas antibakteri dari CFS co-culture dan mono-culture L. plantarum tidak menunjukkan perbedaan signifikan, dengan produksi antibakteri optimal terjadi dalam 24-32 jam inkubasi. Dari hasil ini, disimpulkan bahwa aktivitas antibakteri dari plantaricin dapat lebih diidentifikasi melalui metode broth dilution dan bakteri penginduksi L. fermentum E5, E. faecalis A23, serta L. mesenteroides S2 tidak efektif untuk meningkatkan produksi plantaricin dari L. plantarum SU-KC1a. / Bacteriocins are antimicrobial peptides produced by lactic acid bacteria and have the potential to serve as alternatives of antibiotics and chemical food preservatives. A type of class II bacteriocin known as plantaricin is produced by Lactiplantibacillus plantarum. Several genes which encode plantaricin have been identified in the genome of L. plantarum SU-KC1a, however factor which induced the expression of plantaricin is not yet known. One method to enhance plantaricin production is through co-culture. This study aims to identify the optimal conditions for inducing plantaricin from L. plantarum SU-KC1a through co-culture with three lactic acid bacteria: Enterococcus faecalis A23, Lactobacillus fermentum E5, and Leuconostoc mesenteroides S2. This study also seeks to determine the optimal growth and plantaricin production time through antibacterial activity assays of the Cell-Free Supernatant (CFS) by well diffusion and broth dilution methods. Protein precipitation with ammonium sulfate and identification of plantaricin molecular weight using Tricine SDS-PAGE will also be conducted. The results indicate that the antibacterial activity detected through the well diffusion method is mainly due to organic acids. However, following protein precipitation and antibacterial assay using the broth dilution method, the antibacterial activity was attributed to peptides, likely plantaricin from L. plantarum SU-KC1a. Protein separation using Tricine SDS-PAGE revealed a protein band around 5 kDa, suspected to be plantaricin. Moreover, a comparison of antibacterial activity between CFS from co-culture and mono-culture of L. plantarum showed no significant difference, with optimal antibacterial production occurring within 24-32 hours of incubation. From these results, it can be concluded that the antibacterial activity of plantaricin is more effectively identified through the broth dilution method, and that the inducer bacteria L. fermentum E5, E. faecalis A23, and L. mesenteroides S2 are ineffective in enhancing the production of plantaricin from L. plantarum SU-KC1a.

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