TOTAL PHENOLIC CONTENT, AND BIOLOGICAL POTENTIAL OF THE METHANOLIC EXTRACT FROM THE LICHEN PYRENULA SP.
Main Article Content
Abstract
Pyrenula lichens are highly diverse and widely distributed in Vietnam; however, their chemical composition and biological activities remain largely unexplored. In this study, preliminary screening of major groups of secondary metabolites, the phenolic content, antioxidant, antibacterial, and a-glucosidase inhibitory activities of Pyrenula lichen collected in Ho Chi Minh City were investigated. The presence of alkaloids, terpenoids, coumarins, quinones, tannins, phenols, and flavonoids in the extract was confirmed by qualitative screening. The total phenolic content (TPC) was determined to be 30.26 ± 0.51 mg GAE/g extract. In terms of bioactivities, the extract exhibited moderate antioxidant activity in the DPPH scavenging assay, with an IC50 value of 97.4 ± 8.5 µg/mL. Besides, the alpha-glucosidase inhibition of the extract (IC50 = 46.5 ± 3.6 µg/mL) suggests its potential for antidiabetic management. Moreover, the extract showed strong antibacterial activity against Staphylococcus aureus, confirmed by an inhibition zone diameter of 20.9 ± 0.1 mm and a MIC value of 15.62 μg/mL. These results show that Pyrenula sp. is a promising source for further investigation of biologically active secondary metabolites and their potential applications.
Keywords
antibacterial, antioxidant, α-glucosidase inhibition, Pyrenula sp., total phenolic content
Article Details
References
Acharius, E. (1814). Synopsis methodica lichenum. Litteris et sumptibus Svanborg et soc., Lund, 392 pp.
Aptroot, A. (2012). A world key to the species of Anthracothecium and Pyrenula. The Lichenologist, 44(1), 5–53. https://doi.org/10.1017/S0024282911000624
Bui, A. T., Vo, T. P. G., & Tran, T. M. D. (2024). Alpha-glucosidase inhibitory and antioxidant activities of ethyl acetate extract from Graphis sp. mycobiont. Ho Chi Minh City University of Education Journal of Science, 21(3), 2065–2080. https://doi.org/10.54607/hcmue.js.21.3.4063(2023)
Chan, C. W. S., & Khachigian, L. M. (2013). DNAzyme Delivery Approaches in Biological Settings. http://Www.Eurekaselect.Com, 20(28), 3448–3455. https://doi.org/10.2174/09298673113209990033
Do, T.-H., Duong, T.-H., Nguyen, H. T., Nguyen, T.-H., Sichaem, J., Nguyen, C. H., Nguyen, H.-H., & Long, N. P. (2022). Biological activities of lichen-derived monoaromatic compounds. Molecules, 27(9), Article 9. https://doi.org/10.3390/molecules27092871
Do, T. H., Duong, T. H., Vu, Y. T., Tran, H. P., Nguyen, T. T. N., Sichaem, J., Nguyen, N. H., Nguyen, H. T., & Pham, D.-D. (2024). Alpha-glucosidase inhibitory compounds from Vietnamese lichen Usnea baileyi: In vitro and in silico aspects. RSC Advances, 14(44), 32624–32636. https://doi.org/10.1039/D4RA04449E
Dou, M., Liu, S., Li, J., Aptroot, A., & Jia, Z. (2024). Three new Pyrenula species with 3-septate ascospores with red or orange oil when over-mature (Ascomycota, Pyrenulales, Pyrenulaceae) from China. MycoKeys, 102, 107–125. https://doi.org/10.3897/mycokeys.102.113619
Elshikh, M., Ahmed, S., Funston, S., Dunlop, P., McGaw, M., Marchant, R., & Banat, I. M. (2016). Resazurin-based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants. Biotechnology Letters, 38(6), 1015–1019. https://doi.org/10.1007/s10529-016-2079-2
Fernández-Moriano, C., González-Burgos, E., Divakar, P. K., Crespo, A., & Gómez-Serranillos, M. P. (2016). Evaluation of the Antioxidant Capacities and Cytotoxic Effects of Ten Parmeliaceae Lichen Species. Evidence-Based Complementary and Alternative Medicine, 2016(1), 3169751. https://doi.org/10.1155/2016/3169751
Jeyakumaran, P., Perera, K., Fernando, W., Jayasinghe, L., & Ramiah, S. (2015). α-Glucosidase and α-Amylase Inhibitory Activities of Nine Sri Lankan Antidiabetic Plants. British Journal of Pharmaceutical Research, 7, 365–374. https://doi.org/10.9734/BJPR/2015/18645
Junior, I. O., Aptroot, A., & Cáceres, M. E. da S. (2021). Lichens from Monte Pascoal, Bahia, Brazil, with some new pyrenocarpous species and a key to the Pyrenula species from Brazil. The Bryologist, 124(4), 552–568. https://doi.org/10.1639/0007-2745-124.4.552
Kuete, V. (2010). Potential of Cameroonian plants and derived products against microbial infections: A review. Planta Medica, 76(14), 1479–1491. https://doi.org/10.1055/s-0030-1250027
Le, D. H., Takenaka, Y., Hamada, N., Mizushina, Y., & Tanahashi, T. (2014). Polyketides from the cultured lichen mycobiont of a Vietnamese Pyrenula sp. Journal of Natural Products, 77(6), 1404–1412. https://doi.org/10.1021/np500143k
Lücking, R., Álvaro-Alba, W. R., Moncada, B., Marín-Canchala, N. L., Tunjano, S. S., & Cárdenas-López, D. (2023). Lichens from the Colombian Amazon: 666 Taxa Including 28 new Species and 157 New Country Records Document an Extraordinary Diversity. The Bryologist, 126(2), 242–303. https://doi.org/10.1639/0007-2745-126.2.242
Miranda-González, R., Bungartz, F., Lücking, R., Gaya, E., Mendonça, C. de O., Viñas-Portilla, C., Cáceres, M. E. da S., & Herrera-Campos, M. de los A. (2022). Phylogeny of the Pyrenula ochraceoflava group (Pyrenulaceae) reveals near-cryptic diversification and the inclusion of the Mazaediothecium album aggregate. The Bryologist, 125(4), 541–557. https://doi.org/10.1639/0007-2745-125.4.541
Nguyen, T. D., Nguyen, H. L., Pham, T. P. V., Vo, T. P. G., & Tran, T. M. D. (2024). Impact of culture media and duration on antioxidant and alpha-glucosidase inhibitory activities of methanol extract from Graphis handelii mycobiont. Ho Chi Minh City University of Education Journal of Science, 21(9), 9. https://doi.org/10.54607/hcmue.js.21.9.4199(2024)
Pant, K., Joshi, P., Upreti, D. K., Nayaka, S., & Darmwal, J. (2025). Phytochemical Investigation, Antioxidant and Antimicrobial Activities of Some Parmeliaceae Lichens from Western Himalaya. Journal of Pure and Applied Microbiology, 19(1), 263–274. https://doi.org/10.22207/JPAM.19.1.17
Phuong, N., Do, N., Tai, N., Phuong, P., & Duc, T. (2017). Khảo sát hàm lượng flavonoid, alkaloid và khả năng kháng khuẩn của cao chiết cỏ Mần Trầu (Eleusine indica) [Investigation of flavonoid and alkaloid contents and antibacterial activity of Eleusine indica extracts]. Can Tho University, Journal of Science, 53, 54. https://doi.org/10.22144/ctu.jvn.2017.157
Pradhan, S., Dash, S., Parida, S., Sahoo, B., & Rath, B. (2023). Antioxidant and antimicrobial activities and GC/MS-based phytochemical analysis of two traditional Lichen species Trypethellium virens and Phaeographis dendritica. Journal of Genetic Engineering and Biotechnology, 21(1), 41. https://doi.org/10.1186/s43141-023-00490-0
Pradhan, S., Dash, S., Sahoo, B., Parida, S., & Rath, B. (2024). Screening and characterization of bioactive compounds from two epiphytic microlichen and evaluation of their in vitro antioxidant activity. Future Journal of Pharmaceutical Sciences, 10(1), 10. https://doi.org/10.1186/s43094-024-00584-7
Proença, C., Freitas, M., Ribeiro, D., Oliveira, E. F. T., Sousa, J. L. C., Tomé, S. M., Ramos, M. J., Silva, A. M. S., Fernandes, P. A., & Fernandes, E. (2017). α-Glucosidase inhibition by flavonoids: An in vitro and in silico structure–activity relationship study. Journal of Enzyme Inhibition and Medicinal Chemistry, 32(1), 1216–1228. https://doi.org/10.1080/14756366.2017.1368503
Rashmi, S., & Rajkumar, H. (2014). Preliminary phytochemical screening of different solvent extracts of lichens from Kodagu district, Karnataka. Journal of Pharmacognosy and Phytochemistry. https://www.semanticscholar.org/paper/Preliminary-phytochemical-screening-of-different-of-Rashmi-Rajkumar/99d8c75616c30b141dbe8bb6441c979820a7aea2
Sargsyan, R., Gasparyan, A., Tadevosyan, G., & Panosyan, H. (2021). Antimicrobial and antioxidant potentials of non-cytotoxic extracts of corticolous lichens sampled in Armenia. AMB Express, 11(1), 110. https://doi.org/10.1186/s13568-021-01271-z
Sharma, B. P., Rani, J. J., Vimala, V., Sable, P. D., S. Kumar, S. K., Marndi, S., & Das, K. (2025). Bioactive Compounds of Medicinal Plants: Structure, Function, and Bioactivity. https://doi.org/10.5281/zenodo.17480458
Soto-Medina, E., Aptroot, A., & Lücking, R. (2023). New Species of Lichen for Colombia Tropical Dry Forest. Cryptogamie, Mycologie, 44(7), 103–107. https://doi.org/10.5252/cryptogamie-mycologie2023v44a7
Takenaka, Y., Tanahashi, T., Nagakura, N., & Hamada, N. (2000). Production of xanthones with free radical scavenging properties, emodin and sclerotiorin by the cultured lichen mycobionts of Pyrenula japonica. Zeitschrift Fur Naturforschung. C, Journal of Biosciences, 55(11–12), 910–914. https://doi.org/10.1515/znc-2000-11-1211
Torres-Benítez, A., Ortega-Valencia, J. E., Jara-Pinuer, N., Sanchez, M., Vargas-Arana, G., Gómez-Serranillos, M. P., & Simirgiotis, M. J. (2023). Antioxidant and antidiabetic activity and phytoconstituents of lichen extracts with temperate and polar distribution. Frontiers in Pharmacology, 14. https://doi.org/10.3389/fphar.2023.1251856
Vo, T. K. N., Nguyen, T. H., Tran, H. D., Nguyen, D. D., & Nguyen, T. N. M. (2017). Khảo sát hàm lượng phenolic tổng, flavonoid tổng, hoạt tính chống oxy hóa và hoạt tính kháng khuẩn của cao chiết ethanol và methanol của lá và thân rễ cây Cỏ Tranh (Imperata cylindrica) [Investigation of total phenolic and flavonoid contents, antioxidant and antibacterial activities of ethanol and methanol extracts from the leaves and rhizomes of Imperata cylindrica]. Can Tho University, Journal of Science, (52), 16–22. https://doi.org/10.22144/ctu.jvn.2017.119
White, P. A. S., Oliveira, R. C. M., Oliveira, A. P., Serafini, M. R., Araújo, A. A. S., Gelain, D. P., Moreira, J. C. F., Almeida, J. R. G. S., Quintans, J. S. S., Quintans-Junior, L. J., & Santos, M. R. V. (2014). Antioxidant Activity and Mechanisms of Action of Natural Compounds Isolated from Lichens: A Systematic Review. Molecules, 19(9), 14496–14527. https://doi.org/10.3390/molecules190914496
Zakeri, Z., Junne, S., Jäger, F., Dostert, M., Otte, V., & Neubauer, P. (2022). Lichen cell factories: Methods for the isolation of photobiont and mycobiont partners for defined pure and co-cultivation. Microbial Cell Factories, 21(1), 80. https://doi.org/10.1186/s12934-022-01804-6
Aptroot, A. (2012). A world key to the species of Anthracothecium and Pyrenula. The Lichenologist, 44(1), 5–53. https://doi.org/10.1017/S0024282911000624
Bui, A. T., Vo, T. P. G., & Tran, T. M. D. (2024). Alpha-glucosidase inhibitory and antioxidant activities of ethyl acetate extract from Graphis sp. mycobiont. Ho Chi Minh City University of Education Journal of Science, 21(3), 2065–2080. https://doi.org/10.54607/hcmue.js.21.3.4063(2023)
Chan, C. W. S., & Khachigian, L. M. (2013). DNAzyme Delivery Approaches in Biological Settings. http://Www.Eurekaselect.Com, 20(28), 3448–3455. https://doi.org/10.2174/09298673113209990033
Do, T.-H., Duong, T.-H., Nguyen, H. T., Nguyen, T.-H., Sichaem, J., Nguyen, C. H., Nguyen, H.-H., & Long, N. P. (2022). Biological activities of lichen-derived monoaromatic compounds. Molecules, 27(9), Article 9. https://doi.org/10.3390/molecules27092871
Do, T. H., Duong, T. H., Vu, Y. T., Tran, H. P., Nguyen, T. T. N., Sichaem, J., Nguyen, N. H., Nguyen, H. T., & Pham, D.-D. (2024). Alpha-glucosidase inhibitory compounds from Vietnamese lichen Usnea baileyi: In vitro and in silico aspects. RSC Advances, 14(44), 32624–32636. https://doi.org/10.1039/D4RA04449E
Dou, M., Liu, S., Li, J., Aptroot, A., & Jia, Z. (2024). Three new Pyrenula species with 3-septate ascospores with red or orange oil when over-mature (Ascomycota, Pyrenulales, Pyrenulaceae) from China. MycoKeys, 102, 107–125. https://doi.org/10.3897/mycokeys.102.113619
Elshikh, M., Ahmed, S., Funston, S., Dunlop, P., McGaw, M., Marchant, R., & Banat, I. M. (2016). Resazurin-based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants. Biotechnology Letters, 38(6), 1015–1019. https://doi.org/10.1007/s10529-016-2079-2
Fernández-Moriano, C., González-Burgos, E., Divakar, P. K., Crespo, A., & Gómez-Serranillos, M. P. (2016). Evaluation of the Antioxidant Capacities and Cytotoxic Effects of Ten Parmeliaceae Lichen Species. Evidence-Based Complementary and Alternative Medicine, 2016(1), 3169751. https://doi.org/10.1155/2016/3169751
Jeyakumaran, P., Perera, K., Fernando, W., Jayasinghe, L., & Ramiah, S. (2015). α-Glucosidase and α-Amylase Inhibitory Activities of Nine Sri Lankan Antidiabetic Plants. British Journal of Pharmaceutical Research, 7, 365–374. https://doi.org/10.9734/BJPR/2015/18645
Junior, I. O., Aptroot, A., & Cáceres, M. E. da S. (2021). Lichens from Monte Pascoal, Bahia, Brazil, with some new pyrenocarpous species and a key to the Pyrenula species from Brazil. The Bryologist, 124(4), 552–568. https://doi.org/10.1639/0007-2745-124.4.552
Kuete, V. (2010). Potential of Cameroonian plants and derived products against microbial infections: A review. Planta Medica, 76(14), 1479–1491. https://doi.org/10.1055/s-0030-1250027
Le, D. H., Takenaka, Y., Hamada, N., Mizushina, Y., & Tanahashi, T. (2014). Polyketides from the cultured lichen mycobiont of a Vietnamese Pyrenula sp. Journal of Natural Products, 77(6), 1404–1412. https://doi.org/10.1021/np500143k
Lücking, R., Álvaro-Alba, W. R., Moncada, B., Marín-Canchala, N. L., Tunjano, S. S., & Cárdenas-López, D. (2023). Lichens from the Colombian Amazon: 666 Taxa Including 28 new Species and 157 New Country Records Document an Extraordinary Diversity. The Bryologist, 126(2), 242–303. https://doi.org/10.1639/0007-2745-126.2.242
Miranda-González, R., Bungartz, F., Lücking, R., Gaya, E., Mendonça, C. de O., Viñas-Portilla, C., Cáceres, M. E. da S., & Herrera-Campos, M. de los A. (2022). Phylogeny of the Pyrenula ochraceoflava group (Pyrenulaceae) reveals near-cryptic diversification and the inclusion of the Mazaediothecium album aggregate. The Bryologist, 125(4), 541–557. https://doi.org/10.1639/0007-2745-125.4.541
Nguyen, T. D., Nguyen, H. L., Pham, T. P. V., Vo, T. P. G., & Tran, T. M. D. (2024). Impact of culture media and duration on antioxidant and alpha-glucosidase inhibitory activities of methanol extract from Graphis handelii mycobiont. Ho Chi Minh City University of Education Journal of Science, 21(9), 9. https://doi.org/10.54607/hcmue.js.21.9.4199(2024)
Pant, K., Joshi, P., Upreti, D. K., Nayaka, S., & Darmwal, J. (2025). Phytochemical Investigation, Antioxidant and Antimicrobial Activities of Some Parmeliaceae Lichens from Western Himalaya. Journal of Pure and Applied Microbiology, 19(1), 263–274. https://doi.org/10.22207/JPAM.19.1.17
Phuong, N., Do, N., Tai, N., Phuong, P., & Duc, T. (2017). Khảo sát hàm lượng flavonoid, alkaloid và khả năng kháng khuẩn của cao chiết cỏ Mần Trầu (Eleusine indica) [Investigation of flavonoid and alkaloid contents and antibacterial activity of Eleusine indica extracts]. Can Tho University, Journal of Science, 53, 54. https://doi.org/10.22144/ctu.jvn.2017.157
Pradhan, S., Dash, S., Parida, S., Sahoo, B., & Rath, B. (2023). Antioxidant and antimicrobial activities and GC/MS-based phytochemical analysis of two traditional Lichen species Trypethellium virens and Phaeographis dendritica. Journal of Genetic Engineering and Biotechnology, 21(1), 41. https://doi.org/10.1186/s43141-023-00490-0
Pradhan, S., Dash, S., Sahoo, B., Parida, S., & Rath, B. (2024). Screening and characterization of bioactive compounds from two epiphytic microlichen and evaluation of their in vitro antioxidant activity. Future Journal of Pharmaceutical Sciences, 10(1), 10. https://doi.org/10.1186/s43094-024-00584-7
Proença, C., Freitas, M., Ribeiro, D., Oliveira, E. F. T., Sousa, J. L. C., Tomé, S. M., Ramos, M. J., Silva, A. M. S., Fernandes, P. A., & Fernandes, E. (2017). α-Glucosidase inhibition by flavonoids: An in vitro and in silico structure–activity relationship study. Journal of Enzyme Inhibition and Medicinal Chemistry, 32(1), 1216–1228. https://doi.org/10.1080/14756366.2017.1368503
Rashmi, S., & Rajkumar, H. (2014). Preliminary phytochemical screening of different solvent extracts of lichens from Kodagu district, Karnataka. Journal of Pharmacognosy and Phytochemistry. https://www.semanticscholar.org/paper/Preliminary-phytochemical-screening-of-different-of-Rashmi-Rajkumar/99d8c75616c30b141dbe8bb6441c979820a7aea2
Sargsyan, R., Gasparyan, A., Tadevosyan, G., & Panosyan, H. (2021). Antimicrobial and antioxidant potentials of non-cytotoxic extracts of corticolous lichens sampled in Armenia. AMB Express, 11(1), 110. https://doi.org/10.1186/s13568-021-01271-z
Sharma, B. P., Rani, J. J., Vimala, V., Sable, P. D., S. Kumar, S. K., Marndi, S., & Das, K. (2025). Bioactive Compounds of Medicinal Plants: Structure, Function, and Bioactivity. https://doi.org/10.5281/zenodo.17480458
Soto-Medina, E., Aptroot, A., & Lücking, R. (2023). New Species of Lichen for Colombia Tropical Dry Forest. Cryptogamie, Mycologie, 44(7), 103–107. https://doi.org/10.5252/cryptogamie-mycologie2023v44a7
Takenaka, Y., Tanahashi, T., Nagakura, N., & Hamada, N. (2000). Production of xanthones with free radical scavenging properties, emodin and sclerotiorin by the cultured lichen mycobionts of Pyrenula japonica. Zeitschrift Fur Naturforschung. C, Journal of Biosciences, 55(11–12), 910–914. https://doi.org/10.1515/znc-2000-11-1211
Torres-Benítez, A., Ortega-Valencia, J. E., Jara-Pinuer, N., Sanchez, M., Vargas-Arana, G., Gómez-Serranillos, M. P., & Simirgiotis, M. J. (2023). Antioxidant and antidiabetic activity and phytoconstituents of lichen extracts with temperate and polar distribution. Frontiers in Pharmacology, 14. https://doi.org/10.3389/fphar.2023.1251856
Vo, T. K. N., Nguyen, T. H., Tran, H. D., Nguyen, D. D., & Nguyen, T. N. M. (2017). Khảo sát hàm lượng phenolic tổng, flavonoid tổng, hoạt tính chống oxy hóa và hoạt tính kháng khuẩn của cao chiết ethanol và methanol của lá và thân rễ cây Cỏ Tranh (Imperata cylindrica) [Investigation of total phenolic and flavonoid contents, antioxidant and antibacterial activities of ethanol and methanol extracts from the leaves and rhizomes of Imperata cylindrica]. Can Tho University, Journal of Science, (52), 16–22. https://doi.org/10.22144/ctu.jvn.2017.119
White, P. A. S., Oliveira, R. C. M., Oliveira, A. P., Serafini, M. R., Araújo, A. A. S., Gelain, D. P., Moreira, J. C. F., Almeida, J. R. G. S., Quintans, J. S. S., Quintans-Junior, L. J., & Santos, M. R. V. (2014). Antioxidant Activity and Mechanisms of Action of Natural Compounds Isolated from Lichens: A Systematic Review. Molecules, 19(9), 14496–14527. https://doi.org/10.3390/molecules190914496
Zakeri, Z., Junne, S., Jäger, F., Dostert, M., Otte, V., & Neubauer, P. (2022). Lichen cell factories: Methods for the isolation of photobiont and mycobiont partners for defined pure and co-cultivation. Microbial Cell Factories, 21(1), 80. https://doi.org/10.1186/s12934-022-01804-6