Bioprospecting Ascidian-Associated Aspergillus sp. from Sigillina sp. in Bunaken as a Source of Bioactive Metabolites: Phytochemical Profiling, Antioxidant–Antimicrobial Activities, and ITS-Based Identification
DOI:
https://doi.org/10.35800/jip.v14i2.68654Keywords:
ascidian, Aspergillus sp, antimicrobial, antioxidant, Bunaken, DPPH, marine-derived fungi, Sigillina sp.Abstract
This study aimed to isolate a symbiotic fungus from the ascidian Sigillina sp. collected from Bunaken waters, evaluate the phytochemical profile of its ethyl acetate extract, determine antioxidant and antimicrobial activities, and identify the isolate using internal transcribed spacer (ITS) sequences. The ascidian sample was collected purposively by SCUBA at 5-13 m depth. Fungal isolation was performed on Potato Dextrose Agar, followed by purification, rice-medium cultivation, ethyl acetate extraction, phytochemical screening, DPPH assay, disc diffusion antimicrobial assay, and ITS-based molecular identification through PCR, sequencing, BLAST, and phylogenetic interpretation. The isolate AFBK 5c produced 0.719 g of dry extract. The extract was positive for phenolics, flavonoids, triterpenoids, tannins, and alkaloids, while steroids and saponins were not detected. DPPH inhibition increased from 51.68% at 25 ppm to 71.19% at 125 ppm, with a linear regression of y = 0.20112x + 48.002 (R² = 0.9590), indicating strong in vitro radical-scavenging capacity. The extract showed selective antimicrobial activity, particularly against Candida albicans and Aeromonas hydrophila, with inhibition zones up to 15 mm. ITS analysis produced a 779 bp sequence showing 100% similarity to Aspergillus sp. These findings indicate that Aspergillus sp. AFBK 5c associated with Sigillina sp. from Bunaken is a promising preliminary source of bioactive metabolites, although claims should remain limited to crude-extract-based in vitro screening.
Keywords: ascidian; Aspergillus sp.; antimicrobial; antioxidant; Bunaken; DPPH; marine-derived fungi; Sigillina sp.
Abstrak. Penelitian ini bertujuan mengisolasi jamur simbion dari ascidia Sigillina sp. asal perairan Bunaken, mengevaluasi profil fitokimia ekstrak etil asetat, menentukan aktivitas antioksidan dan antimikroba, serta mengidentifikasi isolat secara molekuler menggunakan sekuens internal transcribed spacer (ITS). Sampel ascidia dikoleksi secara purposive menggunakan SCUBA pada kedalaman 5-13 m. Isolasi jamur dilakukan pada media Potato Dextrose Agar, dilanjutkan pemurnian isolat, kultur pada media nasi, ekstraksi etil asetat, penapisan fitokimia, uji antioksidan DPPH, uji antimikroba difusi cakram, serta identifikasi molekuler melalui PCR, sekuensing ITS, BLAST, dan interpretasi filogenetik. Isolat AFBK 5c menghasilkan ekstrak kering sebesar 0,719 g. Ekstrak menunjukkan hasil positif terhadap fenolik, flavonoid, triterpenoid, tanin, dan alkaloid, sedangkan steroid dan saponin tidak terdeteksi. Inhibisi DPPH meningkat dari 51,68% pada 25 ppm menjadi 71,19% pada 125 ppm, dengan persamaan regresi y = 0,20112x + 48,002 (R² = 0,9590), sehingga menunjukkan kapasitas peredaman radikal bebas in vitro yang kuat. Aktivitas antimikroba bersifat selektif, terutama terhadap Candida albicans dan Aeromonas hydrophila dengan zona hambat hingga 15 mm. Identifikasi ITS menghasilkan sekuens 779 bp dengan kemiripan 100% terhadap Aspergillus sp. Temuan ini menunjukkan bahwa Aspergillus sp. AFBK 5c simbion Sigillina sp. dari Bunaken berpotensi sebagai sumber awal metabolit bioaktif, dengan klaim yang tetap dibatasi pada skrining in vitro berbasis ekstrak kasar.
Kata kunci: antimikroba; antioksidan; ascidia; Aspergillus sp.; Bunaken; DPPH; jamur simbion; Sigillina sp.
References
Baturo-Cieśniewska, A., Pusz, W., & Patejuk, K. (2020). Problems, limitations, and challenges in species identification of Ascomycota members on the basis of ITS regions. Acta Mycologica, 55(1), 5512. https://doi.org/10.5586/am.5512
Gülçin, İ., & Alwasel, S. H. (2023). DPPH radical scavenging assay. Processes, 11(8), 2248. https://doi.org/10.3390/pr11082248
Hoang, M. T. V., Irinyi, L., Chen, S. C. A., Sorrell, T. C., & Meyer, W. (2019). Dual DNA barcoding for the molecular identification of the agents of invasive fungal infections. Frontiers in Microbiology, 10, 1647. https://doi.org/10.3389/fmicb.2019.01647
Li, H., Fu, Y., & Song, F. (2023). Marine Aspergillus: A treasure trove of antimicrobial compounds. Marine Drugs, 21(5), 277. https://doi.org/10.3390/md21050277
Palit, C., Sumilat, D. A., Rumengan, A. P., Boneka, F. B., Sinjal, C. A. L., & Lalita, J. (2022). Komunitas dan keanekaragaman ascidia di Pesisir Minanga, Malalayang Satu, Kota Manado. Jurnal Pesisir dan Laut Tropis, 10(2), 219-225. https://doi.org/10.35800/jplt.10.2.2022.54995
Pan, C., Hassan, S. S. U., Muhammad, I., & Jin, H. (2025). Marine fungi as a goldmine for novel antibiotics: A 2024 perspective. Frontiers in Marine Science, 11, 1538136. https://doi.org/10.3389/fmars.2024.1538136
Qi, G., Hao, L., Gan, Y., Xin, T., Lou, Q., Xu, W., & Song, J. (2024). Identification of closely related species in Aspergillus through Analysis of Whole-Genome. Frontiers in Microbiology, 15, 1323572. https://doi.org/10.3389/fmicb.2024.1323572
Schoch, C. L., Seifert, K. A., Huhndorf, S., Robert, V., Spouge, J. L., Levesque, C. A., Chen, W., & Fungal Barcoding Consortium. (2012). Nuclear ribosomal internal transcribed spacer region as a universal DNA barcode marker for Fungi. Proceedings of the National Academy of Sciences, 109(16), 6241-6246. https://doi.org/10.1073/pnas.1117018109
Shahidi, F., & Samarasinghe, A. (2025). How to assess antioxidant activity? Advances, limitations, and applications of in vitro, in vivo, and ex vivo approaches. Food Production, Processing and Nutrition, 7, 50. https://doi.org/10.1186/s43014-025-00326-z
Sumilat, D. A., Ginting, E. L., Pollo, G. A. V., Adam, A. A., & Tallei, T. E. (2020). Antimicrobial activities of Rhopalaea-associated fungus Aspergillus flavus strain MFABU9. Pakistan Journal of Biological Sciences, 23(1), 1-6.
Vitale, G. A., Coppola, D., Palma Esposito, F., Buonocore, C., Ausuri, J., Tortorella, E., & de Pascale, D. (2020). Antioxidant molecules from marine fungi: Methodological approaches and perspectives. Marine Drugs, 18(12), 645. https://doi.org/10.3390/md18120645
Wang, B., Cai, J., Huang, L., Chen, Y., Wang, R., Luo, M., Yang, M., Zhang, M., Nasihat, Chen, G., Huang, G., & Zheng, C. (2024). Significance of research on natural products from marine-derived Aspergillus species as a source against pathogenic bacteria. Frontiers in Microbiology, 15, 1464135. https://doi.org/10.3389/fmicb.2024.1464135
Wang, C., Tang, S., & Cao, S. (2021). Antimicrobial compounds from marine fungi. Phytochemistry Reviews, 20(1), 85-117. https://doi.org/10.1007/s11101-020-09705-5
Wang, Z., Zhao, M., Li, C., Yu, Y., Gong, Z., Kong, F., & Li, C. (2025). Recent advances in secondary metabolites from marine Aspergillus. Marine Drugs, 23(10), 400. https://doi.org/10.3390/md23100400
White, T. J., Bruns, T., Lee, S., & Taylor, J. (1990). Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In M. A. Innis, D. H. Gelfand, J. J. Sninsky, & T. J. White (Eds.), PCR Protocols: A Guide to Methods and Applications (pp. 315-322). Academic Press.
Wigati, D., Setyowati, E. P., Pratiwi, S. U. T., & Nugraha, A. S. (2024). Promising sponge-derived marine fungi as antibacterial and biofilm inhibitors. Journal of Applied Pharmaceutical Science, 14(4), 14-34. https://doi.org/10.7324/JAPS.2024.161885
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2026 Deiske Adeliene Sumilat, Rosita Lintang, Grevo S. Gerung, Gian Montolalu, Monika M.O. Caroles
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
COPYRIGHT
Authors who publish with this journal agree to the following terms:
Authors hold their copyright and grant this journal the privilege of first publication, with the work simultaneously licensed under a Creative Commons Attribution License that permits others to impart the work with an acknowledgment of the work's origin and initial publication by this journal.
Authors can enter into separate or additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (for example, post it to an institutional repository or publish it in a book), with an acknowledgment of its underlying publication in this journal.
Authors are permitted and encouraged to post their work online (for example, in institutional repositories or on their website) as it can lead to productive exchanges, as well as earlier and greater citation of the published work (See The Effect of Open Access).
