Extracellular Metabolites of Clove-Endophytic Bacteria, Niallia nealsonii DCL1, Exhibit Potential Cellular Antioxidant and Antiaging Activities

Nadiyah Salsabilah, Muhammad Eka Prastya, Rika Indri Astuti

Abstract


The clove plant, Syzygium aromaticum L., is a highly valued spice in Indonesia, recognized for its extensive applications in food preservation and medicinal uses. In a prior study, six endophytic bacteria were successfully isolated from clove leaves. One of these isolates, designated as DCL1, is a Gram-positive bacterium identified as Niallia nealsonii. This study aimed to determine the antioxidant and antiaging properties of the extracellular metabolite of DCL1. Extracellular metabolites were extracted with ethyl acetate and evaluated for antioxidant activity via the DPPH assay. Total phenolic and flavonoid compounds were also quantified with further metabolilte profiling using Liquid Chromatography Tandem-Mass Spectrometry (LC-MS/MS) analysis. Further study of antiaging activity was assayed using yeast model Schizosacchromyces pombe. The extract from N. nealsonii DCL1 cultures harvested at 28 and 40 hours yielded 0.01% and 0.009%, respectively. Our findings indicate that the 40-hour extract contains higher levels of phenolics (93.54 mg gallic acid equivalents/g extract) and flavonoids (12.09 mg quercetin equivalents/g extract) compared to the 28-hour extract. Additionally, the 40-hour extract exhibited greater antioxidant activity than the 28-hour extract, as demonstrated by DPPH (IC50 222 µg/mL) and ABTS (IC50 511.43 µg/mL) assays. Moreover, the 40-hour extract significantly prolonged the viability of S. pombe, suggesting its potential as an anti-aging agent. LC-MS/MS analysis identified several potential bioactive compounds, including gallic acid and N-carbamoyl-2,3-dihydroxybenzamide, which may contribute to its bioactivity. Ultimately, our study indicates that DCL1 40-h extract showed antiaging potential, which is promising as source and formulation of biopharmaceutical or cosmeceutical products. 


Keywords


antiaging; antioxidant; LC-MS/MS; Niallia nealsonii DCL1; Schizosaccharomyces pombe

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Abubakar, H., Astuti, R. I., Listyowati, S., Batubara, I., & Wahyudi, A. T. (2022). An orange pigment from the marine bacterium Paracoccus haeundaensis SAB E11 as a prospective source of natural antioxidants. Biodiversitas Journal of Biological Diversity, 23(9), 4730–4737. doi: 10.13057/BIODIV/D230940

Afrendi, E., Prastya, M. E., Astuti, R. I., Wahyuni, W. T., & Batubara, I. (2023). Bioactivity of the Ethanol Extract of Clove (Syzygium aromaticum) as Antitoxin. International Journal of Food Science, 2023. doi: 10.1155/2023/3245210

Amini, S., Astuti, R. I., & Batubara, I. (2022). Apoptosis in Yeast is Modulated by Clove Leaf Extract and Eugenol Potentially by Interfering Caspase YCA1. OnLine Journal of Biological Sciences, 22(3), 388–394. doi: 10.3844/ojbsci.2022.388.394

Ariybah, Z. Q., Astuti, R. I., & Listiyowati, S. (2021). Cell Longevity of Yeast Saccharomyces cerevisiae by Clove Bud Extract Treatment May Occur in Sirtuin-Independent Pathway . OnLine Journal of Biological Sciences Original Research Paper, 21(3). doi: 10.3844/ojbsci.2021.320.327

Astuti, R. I., Prastya, M. E., Batubara, I., Budiarti, E., & Ilmiyawati, A. (2021). Antiaging and Antioxidant Bioactivities of Asteraceae Plant Fractions on the Cellular Functions of the Yeast Schizosaccharomyces pombe. Advances in Pharmacological and Pharmaceutical Sciences, 2021, 2119634. doi: 10.1155/2021/2119634

Batubara, I., Astuti, R. I., Prastya, M. E., Ilmiawati, A., Maeda, M., Suzuki, M., Hamamoto, A., & Takemori, H. (2020). The Antiaging Effect of Active Fractions and Ent-11?-Hydroxy-15-Oxo-Kaur-16-En-19-Oic Acid Isolated from Adenostemma lavenia (L.) O. Kuntze at the Cellular Level. Antioxidants, 9(8), 719. doi: 10.3390/antiox9080719

Chung, K. W., & Chung, H. Y. (2019). The effects of calorie restriction on autophagy: Role on aging intervention. In Nutrients (Vol. 11, Issue 12). MDPI AG. doi: 10.3390/nu11122923

Cortés-Rojas, D. F., de Souza, C. R. F., & Oliveira, W. P. (2014). Clove (Syzygium aromaticum): A precious spice. Asian Pacific Journal of Tropical Biomedicine, 4(2), 90–96. doi: 10.1016/S2221-1691(14)60215-X

Fauzya, A. F., Astuti, R. I., & Mubarik, N. R. (2019). Effect of Ethanol-Derived Clove Leaf Extract on the Oxidative Stress Response in Yeast Schizosaccharomyces pombe. International Journal of Microbiology, 2019(2145378), 1–7. doi: 10.1155/2019/2145378

Gupta, R. S., Patel, S., Saini, N., & Chen, S. (2020). Robust demarcation of 17 distinct Bacillus species clades, proposed as novel Bacillaceae genera, by phylogenomics and comparative genomic analyses: description of Robertmurraya kyonggiensis sp. nov. and proposal for an emended genus Bacillus limiting it only to the members of the Subtilis and Cereus clades of species. International Journal of Systematic and Evolutionary Microbiology, 70(11), 5753–5798. doi: 10.1099/IJSEM.0.004475

Haro-González, J. N., Castillo-Herrera, G. A., Martínez-Velázquez, M., & Espinosa-Andrews, H. (2021). Clove Essential Oil (Syzygium aromaticum L. Myrtaceae): Extraction, Chemical Composition, Food Applications, and Essential Bioactivity for Human Health. Molecules, 26(21), 6387. doi: 10.3390/MOLECULES26216387

Hu, L., Chen, X., Han, L., Zhao, L., Miao, C., Huang, X., Chen, Y., Li, P., & Li, Y. (2019). Two new phenazine metabolites with antimicrobial activities from soil-derived Streptomyces species. The Journal of Antibiotics, 72(7), 574–577. doi: 10.1038/S41429-019-0163-2

Jiang, Z., Kempinski, C., & Chappell, J. (2016). Extraction and Analysis of Terpenes/Terpenoids. Current Protocols in Plant Biology, 1(2), 345. doi: 10.1002/CPPB.20024

Kefayati, Z., Motamed, S. M., Shojaii, A., Noori, M., & Ghods, R. (2017). Antioxidant Activity and Phenolic and Flavonoid Contents of the Extract and Subfractions of Euphorbia splendida Mobayen. Pharmacognosy Research, 9(4), 362. doi: 10.4103/PR.PR_12_17

Khan, A. H., Zou, Z., Xiang, Y., Chen, S., & Tian, X. L. (2019). Conserved signaling pathways genetically associated with longevity across the species. Biochimica et Biophysica Acta - Molecular Basis of Disease, 1865(7), 1745–1755. doi: 10.1016/j.bbadis.2018.09.001

Lesmana, D., Andrianto, D., & Astuti, R. I. (2021). Antiaging properties of the ethanol fractions of clove (Syzygium aromaticum l.) bud and leaf at the cellular levels: Study in yeast schizosaccharomyces pombe. Scientia Pharmaceutica, 89(4). doi: 10.3390/scipharm89040045

Li, Z., Zhang, Z., Ren, Y., Wang, Y., Fang, J., Yue, H., Ma, S., & Guan, F. (2021). Aging and age?related diseases: from mechanisms to therapeutic strategies. In Biogerontology. doi: 10.1007/s10522-021-09910-5

Liang, L., Wang, G., Haltli, B., Marchbank, D. H., Stryhn, H., Correa, H., & Kerr, R. G. (2020). Metabolomic Comparison and Assessment of Co-cultivation and a Heat-Killed Inducer Strategy in Activation of Cryptic Biosynthetic Pathways. Journal of Natural Products, 83(9), 2696–2705. doi: 10.1021/ACS.JNATPROD.0C00621/SUPPL_FILE/NP0C00621_SI_001.PDF

Lobo, V., Patil, A., Phatak, A., & Chandra, N. (2010). Free radicals, antioxidants and functional foods: Impact on human health. Pharmacognosy Reviews, 4(8), 118–126. doi: 10.4103/0973-7847.70902

Martemucci, G., Costagliola, C., Mariano, M., D’andrea, L., Napolitano, P., & D’Alessandro, A. G. (2022). Free Radical Properties, Source and Targets, Antioxidant Consumption and Health. Oxygen, 2(2), 48–78. doi: 10.3390/oxygen2020006

Nurcholis, W., Sya’bani Putri, D. N., Husnawati, H., Aisyah, S. I., & Priosoeryanto, B. P. (2021). Total flavonoid content and antioxidant activity of ethanol and ethyl acetate extracts from accessions of Amomum compactum fruits. Annals of Agricultural Sciences, 66(1), 58–62. doi: 10.1016/J.AOAS.2021.04.001

Pan, Y., Schroeder, E. A., Ocampo, A., Barrientos, A., & Shadel, G. S. (2011). Regulation of yeast chronological life span by TORC1 via adaptive mitochondrial ROS signaling. Cell Metabolism, 13(6), 668–678. doi: 10.1016/j.cmet.2011.03.018

Patel, S., & Gupta, R. S. (2019). A phylogenomic and comparative genomic framework for resolving the polyphyly of the genus bacillus: Proposal for six new genera of bacillus species, peribacillus gen. nov., cytobacillus gen. nov., mesobacillus gen. nov., neobacillus gen. nov., metabacillu. International Journal of Systematic and Evolutionary Microbiology, 70(1), 406–438. doi: 10.1099/ijsem.0.003775

Pinta?, D., Majki?, T., Torovi?, L., Or?i?, D., Beara, I., Simin, N., Mimica–Duki?, N., & Lesjak, M. (2018). Solvent selection for efficient extraction of bioactive compounds from grape pomace. Industrial Crops and Products, 111, 379–390. doi: 10.1016/J.INDCROP.2017.10.038

Pitt, J. N., & Kaeberlein, M. (2015). Why Is Aging Conserved and What Can We Do about It? PLoS Biology, 13(4), 1–11. doi: 10.1371/journal.pbio.1002131

Pizzino, G., Irrera, N., Cucinotta, M., Pallio, G., Mannino, F., Arcoraci, V., Squadrito, F., Altavilla, D., & Bitto, A. (2017). Oxidative Stress: Harms and Benefits for Human Health. Oxidative Medicine and Cellular Longevity, 2017, 1–13. doi: 10.1155/2017/8416763

Plata-Rueda, A., Campos, J. M., da Silva Rolim, G., Martínez, L. C., Dos Santos, M. H., Fernandes, F. L., Serrão, J. E., & Zanuncio, J. C. (2018). Terpenoid constituents of cinnamon and clove essential oils cause toxic effects and behavior repellency response on granary weevil, Sitophilus granarius. Ecotoxicology and Environmental Safety, 156, 263–270. doi: 10.1016/J.ECOENV.2018.03.033

Prastya, M.E., Astuti, R. I., Batubara, I., & Wahyudi, A. T. (2018). Bacillus sp. SAB E-41-derived extract shows antiaging properties via ctt1-mediated oxidative stress tolerance response in yeast Schizosaccharomyces pombe. Asian Pacific Journal of Tropical Biomedicine, 8(11), 533–539. doi: 10.4103/2221-1691.245958

Prastya, Muhammad Eka, Astuti, R. I., Batubara, I., Takagi, H., & Wahyudi, A. T. (2020). Natural extract and its fractions isolated from the marine bacterium Pseudoalteromonas flavipulchra STILL-33 have antioxidant and antiaging activities in Schizosaccharomyces pombe. FEMS Yeast Research, 20(3), 1–14. doi: 10.1093/femsyr/foaa014

Rahmi, D., Yunilawati, R., Jati, B. N., Setiawati, I., Riyanto, A., Batubara, I., & Astuti, R. I. (2021). Antiaging and Skin Irritation Potential of Four Main Indonesian Essential Oils. Cosmetics 2021, Vol. 8, Page 94, 8(4), 94. doi: 10.3390/COSMETICS8040094

Ren, C. Z., Hu, W. Y., Li, J. C., Xie, Y. H., Jia, N. N., Shi, J., Wei, Y. Y., & Hu, T. J. (2020). Ethyl acetate fraction of flavonoids from Polygonum hydropiper L. modulates pseudorabies virus-induced inflammation in RAW264.7 cells via the nuclear factor-kappa B and mitogen-activated protein kinase pathways. The Journal of Veterinary Medical Science, 82(12), 1781. doi: 10.1292/JVMS.20-0263

Riptanti, E. W., Qonita, A., & Uchyani, R. (2019). Revitalization of cloves cultivation in Central Java, Indonesia. IOP Conference Series: Earth and Environmental Science, 314(1), 012085. doi: 10.1088/1755-1315/314/1/012085

Robert, G., & Wagner, J. R. (2020). ROS-Induced DNA Damage as an Underlying Cause of Aging. Advances in Geriatric Medicine and Research. doi: 10.20900/AGMR20200024

Ruetenik, A., & Barrientos, A. (2015). Dietary restriction, mitochondrial function and aging: From yeast to humans. Biochimica et Biophysica Acta - Bioenergetics, 1847(11), 1434–1447. doi: 10.1016/j.bbabio.2015.05.005

Saeedi, P., Petersohn, I., Salpea, P., Malanda, B., Karuranga, S., Unwin, N., Colagiuri, S., Guariguata, L., Motala, A. A., Ogurtsova, K., Shaw, J. E., Bright, D., & Williams, R. (2019). Global and regional diabetes prevalence estimates for 2019 and projections for 2030 and 2045: Results from the International Diabetes Federation Diabetes Atlas, 9th edition. Diabetes Research and Clinical Practice, 157. doi: 10.1016/J.DIABRES.2019.107843

Shahidi, F., & Ambigaipalan, P. (2015). Phenolics and polyphenolics in foods, beverages and spices: Antioxidant activity and health effects - A review. Journal of Functional Foods, 18. doi: 10.1016/j.jff.2015.06.018

Singh, M., Kumar, A., Singh, R., & Pandey, K. D. (2017). Endophytic bacteria: a new source of bioactive compounds. 3 Biotech, 7(315), 1–14. doi: 10.1007/s13205-017-0942-z

Stobiecka, M., Król, J., & Brodziak, A. (2022). Antioxidant Activity of Milk and Dairy Products. In Animals (Vol. 12, Issue 3). doi: 10.3390/ani12030245

Sugiyama, Y., & Hirota, A. (2009). New Potent DPPH Radical Scavengers from a Marine-Derived Actinomycete Strain USF-TC31. Bioscience, Biotechnology, and Biochemistry, 73(12), 2731–2734. doi: 10.1271/BBB.90636

Tan, B. L., Norhaizan, M. E., Liew, W. P. P., & Rahman, H. S. (2018). Antioxidant and oxidative stress: A mutual interplay in age-related diseases. Frontiers in Pharmacology, 9(1162), 1–28. doi: 10.3389/fphar.2018.01162

Thavamoney, N., Sivanadian, L., Tee, L. H., Khoo, H. E., Prasad, K. N., & Kong, K. W. (2018). Extraction and recovery of phytochemical components and antioxidative properties in fruit parts of Dacryodes rostrata influenced by different solvents. Journal of Food Science and Technology, 55(7), 2523. doi: 10.1007/S13197-018-3170-6

Truong, D. H., Ta, N. T. A., Pham, T. V., Huynh, T. D., Do, Q. T. G., Dinh, N. C. G., Dang, C. D., Nguyen, T. K. C., & Bui, A. V. (2021). Effects of solvent—solvent fractionation on the total terpenoid content and in vitro anti?inflammatory activity of Serevenia buxifolia bark extract. Food Science & Nutrition, 9(3), 1720. doi: 10.1002/FSN3.2149

Utami, L. A., Wahyuni, W. T., Mubarik, N. R., & Astuti, R. I. (2023). Endophytic bacteria of clove (Syzygium aromaticum L.) leaves produce metabolites with antioxidant and anti-aging properties. Journal of Applied Pharmaceutical Science, 13,(7), 241–250. doi: 10.7324/JAPS.2023.93258

Vaiserman, A., Koliada, A., Zayachkivska, A., & Lushchak, O. (2020). Nanodelivery of Natural Antioxidants: An Anti-aging Perspective. Frontiers in Bioengineering and Biotechnology, 7. doi: 10.3389/FBIOE.2019.00447

Wierman, M. B., & Smith, J. S. (2014). Yeast sirtuins and the regulation of aging. FEMS Yeast Research, 14(1), 73–88. doi: 10.1111/1567-1364.12115

Xu, D. P., Li, Y., Meng, X., Zhou, T., Zhou, Y., Zheng, J., Zhang, J. J., & Li, H. Bin. (2017). Natural antioxidants in foods and medicinal plants: Extraction, assessment and resources. International Journal of Molecular Sciences, 18(1). doi: 10.3390/ijms18010096

Yang, W., & Hekimi, S. (2010). A Mitochondrial Superoxide Signal Triggers Increased Longevity in Caenorhabditis elegans. PLoS Biology, 8(12), e1000556. doi: 10.1371/journal.pbio.1000556

Yusuf, S. M., Astuti, R. I., Batubara, I., & Chavasiri, W. (2021). Anti-Aging Activity of Xylocarpus Granatum Phytoextracts and Xyloccensins K Compound. Indonesian Journal of Pharmacy, 32(3), 365–375. doi: 10.22146/IJP.1430




DOI: https://doi.org/10.14421/biomedich.2025.142.747-755

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Biology, Medicine, & Natural Product Chemistry
ISSN 2089-6514 (paper) - ISSN 2540-9328 (online)
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