Antimicrobial Activity and Mode of Action of Phellinus hartigii Extract Investigated on Multidrug-Resistant Staphylococcus aureus Using Fourier-Transform Infrared Spectroscopy

Yıl 2025, Cilt: 25 Sayı: 3 , 409 - 421 , 25.12.2025

Kaan Akçay , Kübra Tekşen , Umay Merve Şenturan , Eda Altınöz , Dilşad Özerkan , İlgaz Akata , Ergin Murat Altuner

https://doi.org/10.17475/kastorman.1845436

https://izlik.org/JA33SN87KX

Öz

Aim of study: Macrofungi have garnered attention as promising reservoirs for novel antibiotics. Phellinus hartigii is known to have antimicrobial properties against various microorganisms. This study aims to investigate the in vitro mode of action of the ethanol extract of P. hartigii (PH-EtOH) against multidrug-resistant (MDR) Staphylococcus aureus using Fourier-transform infrared (FTIR) spectroscopy.
Material and method: The composition of the extract was determined by GC-MS analysis, and its antimicrobial activity was evaluated through MIC and MBC tests. The effect of the extract was further analysed using FTIR spectroscopy.
Main results: The PH-EtOH extract demonstrated antibacterial activity with a MIC of 53 µg/mL and an MBC of 420 µg/mL. The major constituents of PH-EtOH were ethyl oleate, linoleic acid ethyl ester, and hexadecanoic acid ethyl ester. FTIR spectra showed changes mainly in proteins and nucleic acids.
Research highlights: The results of this study provide a starting point for further research into the use of P. hartigii ethanol extract as an antimicrobial agent against MDR S. aureus.

Anahtar Kelimeler

Phellinus hartigii , Staphylococcus aureus , Antimicrobial Activity , Mode of Action , FTIR

Destekleyen Kurum

This work was supported by the Kastamonu University Scientific Research Coordination Unit, with project number KÜ-HIZDES/2023-51.

Teşekkür

The authors thank Prof. Dr. Turhan KÖPRÜBAŞI for his valuable guidance in interpreting the zero-order and second-derivative spectra.

Çoklu İlaca Dirençli Staphylococcus aureus Üzerinde Phellinus hartigii Ekstraktının Antimikrobiyal Aktivitesi ve Etki Mekanizmasının Fourier Dönüşüm Infrared Spektroskopisi Kullanılarak İncelenmesi

https://izlik.org/JA33SN87KX

Öz

Çalışmanın amacı: Makrofunguslar, yeni antibiyotikler için umut vadeden rezervuarlar olarak dikkat çekmektedir. Phellinus hartigii’nin çeşitli mikroorganizmalara karşı güçlü antimikrobiyal özellikler taşıdığı gösterilmiştir. Bu çalışma, P. hartigii'nin etanol ekstraktının (PH-EtOH) çoklu ilaca dirençli (MDR) Staphylococcus aureus'a karşı in vitro etki şekline ilişkin FTIR tabanlı kanıt sağlamayı amaçlamaktadır.
Materyal ve yöntem: PH-EtOH'nin bileşimi GC-MS analizi kullanılarak belirlenmiş ve S. aureus'a karşı antimikrobiyal aktivitesi MİK ve MBK testleri ile değerlendirilmiştir. Ekstraktın etkisi FTIR spektroskopisi kullanılarak analiz edilmiştir.
Temel sonuçlar: PH-EtOH ekstraktı, S. aureus'a üzerinde 53 µg/mL'lik bir MİK ve 420 µg/mL'lik bir MBK ile antibakteriyel aktivite göstermiştir. PH-EtOH'nin ana bileşenleri etil oleat, linoleik asit etil ester ve heksadekanoik asit etil ester olarak bulunmuştur. FTIR spektrumları, başlıca protein ve nükleik asitlerde değişiklikler göstermiştir.
Araştırma vurguları: Bu çalışmanın sonuçları, P. hartigii etanol özütünün MDR S. aureus'a karşı antimikrobiyal ajan olarak kullanımına yönelik daha ileri araştırmalar için bir başlangıç noktası sağlamaktadır.

Anahtar Kelimeler

Phellinus hartigii , Staphylococcus aureus , Antimikrobiyal Aktivite , Etki Mekanizması , FTIR

Kaynakça

• Abebe, A. A. & Birhanu, A. G. (2023). Methicillin resistant Staphylococcus aureus: molecular mechanisms underlying drug resistance development and novel strategies to Combat. Infection and Drug Resistance, 7641-7662.
• Adamou, A., Manos, G., Messios, N., Georgiou, L., Xydas, C. & Varotsis, C. (2016). Probing the whole ore chalcopyrite–bacteria interactions and jarosite biosynthesis by Raman and FTIR microspectroscopies. Bioresource technology, 214, 852-855.
• Akpi, U. K., Odoh, C. K., Ideh, E. E. & Adobu, U. S. (2017). Antimicrobial activity of Lycoperdon perlatum whole fruit body on common pathogenic bacteria and fungi. African Journal of Clinical and Experimental Microbiology, 18(2), 79-85.
• Alanis, A. J. (2005). Resistance to antibiotics: are we in the post-antibiotic era?. Archives of medical research, 36(6), 697-705.
• Altuner, E. M. & Akata, I. (2010). Bazı Makromantar Ekstraktlarının Antimikrobiyal Aktivitesi. Sakarya University Journal of Science, 14(1), 45-49.
• Altuner, E. M. & Canlı, K. (2012). In vitro antimicrobial screening of Hypnum andoi AJE Sm. Kastamonu University Journal of Forestry Faculty, 12(1), 97-101.
• Altuner, E. M. & Çetin, B. (2009). Antimicrobial activity of Thuidium delicatulum (Bryopsida) extracts. Kafkas Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 2(2), 85-92.
• Altuner, E. M. & Çetin, B. (2018). Antimicrobial activity of Isothecium alopecuroides and potential effect of some climate elements on the activity of this bryophyte sample. Kastamonu University Journal of Forestry Faculty, 18(2), 126-137.
• Alvarez-Ordóñez, A., Mouwen, D. J. M., López, M. & Prieto, M. (2011). Fourier transform infrared spectroscopy as a tool to characterize molecular composition and stress response in foodborne pathogenic bacteria. Journal of microbiological methods, 84(3), 369-378.
• Appiah, T., Boakye, Y. D. & Agyare, C. (2017). Antimicrobial Activities and Time‐Kill Kinetics of Extracts of Selected Ghanaian Mushrooms. Evidence‐Based Complementary and Alternative Medicine, 2017(1), 4534350.
• Azeem, U., Dhingra, G. S. & Shri, R. (2018). Pharmacological potential of wood inhabiting fungi of genus Phellinus Quél.: An overview. Journal of Pharmacognosy and Phytochemistry, 7(2), 1161-1171.
• Baker, M. J., Trevisan, J., Bassan, P., Bhargava, R., Butler, H. J. & et al. (2014). Using Fourier transform IR spectroscopy to analyze biological materials. Nature protocols, 9(8), 1771-1791.
• Bala, N., Aitken, E. A., Cusack, A. & Steadman, K. J. (2012). Antimicrobial potential of Australian macrofungi extracts against foodborne and other pathogens. Phytotherapy Research, 26(3), 465-469.
• Barth, A. (2007). Infrared spectroscopy of proteins. Biochimica et Biophysica Acta (BBA)-Bioenergetics, 1767(9), 1073-1101.
• Breitenbach, J., & Kränzlin, F. (1986). Champignons de Suisse, champignons sans lames, Tome II. Mykologia, Luzern, Switzerland.
• Canli, K., Bozyel, M. E., Benek, A., Yetgin, A., Senturan, M. & Altuner, E. M. (2020). Chemical composition and in vitro antimicrobial activity of Matthiola tricuspidata ethanol extract. Fresenius Environmental Bulletin, 29(10), 8863-8868.
• Canli, K., Cetin, B., Altuner, E. M., Turkmen, Y., Uzek, U. & Dursun, H. (2014). In vitro antimicrobial screening of Hedwigia ciliata var. leucophaea and determination of the ethanol extract composition by gas chromatography/mass spectrometry (GC/MS). Mass Spectrometry (GC/MS). Journal of Pure and Applied Microbiology, 8(4), 2987-2998.
• Canli, K., Yetgin, A., Akata, I. & Altuner, E. M. (2016a). In vitro antimicrobial activity of Angelica sylvestris roots. International Journal of Biological Sciences, 1(1), 1-7.
• Canli, K., Yetgin, A., Akata, I. & Altuner, E. M. (2016b). In vitro antimicrobial activity screening of Rheum rhabarbarum roots. International Journal of Pharmaceutical Science Invention, 5(2), 01-04.
• Cantón, R., Horcajada, J. P., Oliver, A., Garbajosa, P. R. & Vila, J. (2013). Inappropriate use of antibiotics in hospitals: the complex relationship between antibiotic use and antimicrobial resistance. Enfermedades infecciosas y microbiologia clinica, 31, 3-11.
• Christina, A., Christapher, V. & Bhore, S. J. (2013). Endophytic bacteria as a source of novel antibiotics: an overview. Pharmacognosy reviews, 7(13), 11.
• Cragg, G. M. & Newman, D. J. (2013). Natural products: a continuing source of novel drug leads. Biochimica et Biophysica Acta (BBA)-General Subjects, 1830(6), 3670-3695.
• De Silva, D. D., Rapior, S., Sudarman, E., Stadler, M., Xu, J., Aisyah Alias, S. & Hyde, K. D. (2013). Bioactive metabolites from macrofungi: ethnopharmacology, biological activities and chemistry. Fungal Diversity, 62, 1-40.
• Dousseau, F. & Pezolet, M. (1990). Determination of the secondary structure content of proteins in aqueous solutions from their amide I and amide II infrared bands. Comparison between classical and partial least-squares methods. Biochemistry, 29(37), 8771-8779.
• Faghihzadeh, F., Anaya, N. M., Schifman, L. A. & Oyanedel-Craver, V. (2016). Fourier transform infrared spectroscopy to assess molecular-level changes in microorganisms exposed to nanoparticles. Nanotechnology for Environmental Engineering, 1, 1-16.
• Filip, Z., Herrmann, S. & Kubat, J. (2004). FT-IR spectroscopic characteristics of differently cultivated Bacillus subtilis. Microbiological research, 159(3), 257-262.
• Fjell, C. D., Hiss, J. A., Hancock, R. E. & Schneider, G. (2012). Designing antimicrobial peptides: form follows function. Nature Reviews Drug Discovery, 11(1), 37-51.
• Friedman, N. D., Temkin, E. & Carmeli, Y. (2016). The negative impact of antibiotic resistance. Clinical microbiology and infection, 22(5), 416-422.
• Gupta, K. K. & Rana, D. (2021). Spectroscopic and chromatographic identification of bioprospecting bioactive compounds from cow feces: Antimicrobial and antioxidant activities evaluation of gut bacterium Pseudomonas aeruginosa KD155. Biotechnology Reports, 29, e00577.
• Harvey, A. (2000). Strategies for discovering drugs from previously unexplored natural products. Drug discovery today, 5(7), 294-300.
• Hay, S. I., Rao, P. C., Dolecek, C., Day, N. P., Stergachis, A., Lopez, A. D. & Murray, C. J. (2018). Measuring and mapping the global burden of antimicrobial resistance. BMC medicine, 16, 1-3.
• Högberg, L. D., Heddini, A. & Cars, O. (2010). The global need for effective antibiotics: challenges and recent advances. Trends in pharmacological sciences, 31(11), 509-515.
• Ishida, K. P. & Griffiths, P. R. (1993). Comparison of the amide I/II intensity ratio of solution and solid-state proteins sampled by transmission, attenuated total reflectance, and diffuse reflectance spectrometry. Applied spectroscopy, 47(5), 584-589.
• Li, B. & Webster, T. J. (2018). Bacteria antibiotic resistance: New challenges and opportunities for implant‐associated orthopedic infections. Journal of Orthopaedic Research, 36(1), 22-32.
• Liauw, C. M., Vaidya, M., Slate, A. J., Hickey, N. A., Ryder, S. & et al. (2023). Analysis of cellular damage resulting from exposure of bacteria to graphene oxide and hybrids using fourier transform infrared spectroscopy. Antibiotics, 12(4), 776.
• Lorenz-Fonfria, V. A. (2020). Infrared difference spectroscopy of proteins: from bands to bonds. Chemical reviews, 120(7), 3466-3576.
• MacGowan, A. & Macnaughton, E. (2017). Antibiotic resistance. Medicine, 45(10), 622-628.
• Madejová, J. J. V. S. (2003). FTIR techniques in clay mineral studies. Vibrational spectroscopy, 31(1), 1-10.
• Maity, J. P., Kar, S., Lin, C. M., Chen, C. Y., Chang, Y. F. & et al. (2013). Identification and discrimination of bacteria using Fourier transform infrared spectroscopy. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 116, 478-484.
• Morath, S. U., Hung, R. & Bennett, J. W. (2012). Fungal volatile organic compounds: a review with emphasis on their biotechnological potential. Fungal biology reviews, 26(2-3), 73-83.
• Movasaghi, Z., Rehman, S. & ur Rehman, D. I. (2008). Fourier transform infrared (FTIR) spectroscopy of biological tissues. Applied Spectroscopy Reviews, 43(2), 134-179.
• Muntean, C. M., Stefan, R., Bindea, M. & Cozma, V. (2013). Fourier transform infrared spectroscopy of DNA from Borrelia burgdorferi sensu lato and Ixodes ricinus ticks. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 110, 185-192.
• Muntean, C. M., Ştefan, R., Tabaran, A., Tripon, C., Bende, A. & et al. (2021). The influence of UV femtosecond laser pulses on bacterial DNA structure, as proved by Fourier Transform Infrared (FT‐IR) spectroscopy. ChemistrySelect, 6(27), 6957-6972.
• Nelson, D. W., Millar, B. C., Rao, J. R. & Moore, J. E. (2021). The role of plants and macrofungi as a source of novel antimicrobial agents. Reviews and Research in Medical Microbiology, 32(4), 231-236.
• Nweze, J. A., Mbaoji, F. N., Huang, G., Li, Y., Yang, L. & et al. (2020). Antibiotics development and the potentials of marine-derived compounds to stem the tide of multidrug-resistant pathogenic bacteria, fungi, and protozoa. Marine Drugs, 18(3), 145.
• Ribeiro da Cunha, B., Fonseca, L. P. & Calado, C. R. (2020). Metabolic fingerprinting with Fourier-transform infrared (FTIR) spectroscopy: towards a high-throughput screening assay for antibiotic discovery and mechanism-of-action elucidation. Metabolites, 10(4), 145.
• Sebben, D. & Pendleton, P. (2014). Infrared spectrum analysis of the dissociated states of simple amino acids. Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 132, 706-712.
• Shen, H. S., Shao, S., Chen, J. C. & Zhou, T. (2017). Antimicrobials from mushrooms for assuring food safety. Comprehensive Reviews in Food Science and Food Safety, 16(2), 316-329. Slama, T. G. (2008). Gram-negative antibiotic resistance: there is a price to pay. Critical Care, 12, 1-7.
• Soleimani, Y., Mohammadi, M. R., Schaffie, M., Zabihi, R. & Ranjbar, M. (2023). An experimental study of the effects of bacteria on asphaltene adsorption and wettability alteration of dolomite and quartz. Scientific Reports, 13(1), 21497.
• Spellberg, B., Guidos, R., Gilbert, D., Bradley, J., Boucher, H. W. & et al. (2008). The epidemic of antibiotic-resistant infections: a call to action for the medical community from the Infectious Diseases Society of America. Clinical infectious diseases, 46(2), 155-164.
• Subramani, R., Narayanasamy, M. & Feussner, K. D. (2017). Plant-derived antimicrobials to fight against multi-drug-resistant human pathogens. 3 Biotech, 7, 1-15.
• Surewicz, W. K., Mantsch, H. H. & Chapman, D. (1993). Determination of protein secondary structure by Fourier transform infrared spectroscopy: a critical assessment. Biochemistry, 32(2), 389-394.
• Talari, A. C. S., Martinez, M. A. G., Movasaghi, Z., Rehman, S. & Rehman, I. U. (2017). Advances in Fourier transform infrared (FTIR) spectroscopy of biological tissues. Applied Spectroscopy Reviews, 52(5), 456-506.
• Tenover, F. C. (2006). Mechanisms of antimicrobial resistance in bacteria. The American journal of medicine, 119(6), S3-S10.
• Tsilo, P. H., Basson, A. K., Ntombela, Z. G., Dlamini, N. G. & Pullabhotla, R. V. (2024). Green synthesis and characterisation of silver nanoparticles utilising a bioflocculant obtained from Pichia kudriavzevii isolated from kombucha tea SCOBY. Advances in Materials and Processing Technologies, 1-17.
• URL-1. (2023). R Core Team. R; https://www.R-project.org/. (accessed 14.01.2025).
• Wien, F., Geinguenaud, F., Grange, W. & Arluison, V. (2021). SRCD and FTIR spectroscopies to monitor protein-induced nucleic acid remodeling. RNA Remodeling Proteins: Methods and Protocols, 87-108.
• Zengin Köksal, H., Özerkan, D., Altuner, E. M. & Canlı, K. (2021). Syntrichia ruraliformis (Besch.) Mans., Hypnum andoi AJE Sm. ve Platyhypnidium riparioides Dixon Etanol Ekstraktlarının HCT116 Hücre Canlılığı Üzerindeki Etkilerinin Spektroskopik Açıdan İncelenmesi. Anatolian Bryology, 7(2), 109-118.
• Zhizhina, G. P. & Oleinik, E. F. (1972). Infrared spectroscopy of nucleic acids. Russian Chemical Reviews, 41(3), 258.
• Zhong, J. J. & Xiao, J. H. (2009). Secondary metabolites from higher fungi: discovery, bioactivity, and bioproduction. Biotechnology in China I: from bioreaction to bioseparation and bioremediation, 79-150.