Extraction of Sideritis brevidens P.H. Davis by green synthesis method: determination of antimicrobial and antibiofilm activities against multidrug-resistant bacteria

Yıl 2025, Cilt: 18 Sayı: 3, 494 - 505, 22.12.2025

Mehmet Çimentepe , Özge Öztürk Çimentepe

https://doi.org/10.26559/mersinsbd.1795317

Öz

Aim: This study aimed to investigate the aerial extracts of Sideritis brevidens using supercritical CO2 (ScCO2) extraction and Soxhlet extraction methods, using different solvents, to characterize their phytochemical composition and to evaluate their antimicrobial and antibiofilm activities against multidrug-resistant bacteria. Method: The phytochemical contents of extracts obtained using different methods were analyzed using LC-MS/MS. The antibacterial activities of the extracts against carbapenem-resistant Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae, as well as methicillin-resistant Staphylococcus aureus strains, were investigated using the microdilution method. Furthermore, the antibiofilm activities of the extracts against Acinetobacter baumannii were evaluated using the crystal violet method. Results: Most of the important phenolic compounds obtained from Sideritis brevidens extracts obtained using different methods were identified using the ScCO2 extraction method. Gallic acid and quercetin were found only in the ScCO2-100 and ScCO2-200 extracts. In contrast, chlorogenic acid was found in high amounts in the ethanolic and aqueous extracts. The ScCO2-200 extract was determined the lowest MIC value at a concentration of 16 µg/mL among the tested strains, especially against Acinetobacter baumannii and methicillin-resistant Staphylococcus aureus strains. The ScCO2-200 extract was detected the lowest MIC value at a concentration of 16 µg/mL among the tested strains, especially against Acinetobacter baumannii and methicillin-resistant Staphylococcus aureus strains. According to the disk diffusion method, the largest zone diameter (22 mm) was observed against Acinetobacter baumannii in the ScCO2-100 and ScCO2-200 extracts. In addition, the extract with ScCO2-100 and ScCO2-200 exhibited significant concentration-dependent antibiofilm activity against Acinetobacter baumannii strain. Conclusion: These findings highlight the significant potential of as a natural source of antimicrobial and antibiofilm agents against multidrug-resistant bacterial infections. ScCO2 extraction offers significant advantages in the selective extraction of bioactive phytochemicals.

Anahtar Kelimeler

Sideritis brevidens , supercritical CO2 extraction , soxhlet extraction , antimicrobial and antibiofilm

Sideritis brevidens P.H. Davis türünün yeşil sentez yöntemi ile ekstraksiyonu: çoklu ilaç direnci gösteren bakterilere karşı antimikrobiyal ve antibiyofilm aktivitelerinin belirlenmesi

Öz

Amaç: Bu çalışmada, süperkritik CO2 (ScCO2) ekstraksiyon ve farklı çözücüler kullanılarak soxhlet ekstraksiyon yöntemlerinden elde edilen Sideritis brevidens’in toprak üstü ekstrelerini incelemeyi, fitokimyasal bileşimlerini karakterize etmeyi ve çoklu ilaç dirençli bakterilere karşı antimikrobiyal ve antibiyofim aktivitelerini değerlendirilmesi amaçlanmıştır. Yöntem: Farklı yöntemler ile elde edilen ekstrelerin fitokimyasal içeriği LC-MS/MS tekniği kullanılarak analiz edilmiştir. Ekstrelerin karbapenem dirençli Acinetobacter baumannii, Escherichia coli ve Klebsiella pneumoniae ile metisillin dirençli Staphylococcus aureus suşlarına karşı antibakteriyel etkinlikleri mikrodilüsyon yöntemi araştırılmıştır. Ayrıca, ekstrelerin Acinetobacter baumannii suşuna karşı antibiyofilm aktiviteleri kristal viyole yöntemi ile değerlendirilmiştir. Bulgular: Sideritis brevidens’in farklı yöntemler ile elde edilen ekstrelerinden elde edilen önemli fenolik bileşiklerin çoğu, ScCO2 ekstraksiyon yöntemi kullanılarak tespit edilmiştir. Gallik asit ve kuersetin sadece ScCO2-100 ve ScCO2-200'lü ekstrelerinde bulunmuştur. Buna karşın, klorojenik asit etanollü ve sulu ekstresinde yüksek miktarlarda bulunmuştur. Test edilen suşlar arasında ScCO2-200 ekstresinin 16 µg/mL konsantrasyonunda, özellikle Acinetobacter baumannii ve metisiline dirençli Staphylococcus aureus suşlarına karşı en düşük MİK değeri tespit edilmiştir. Disk difüzyon yöntemine göre, ScCO2-100 ve ScCO2-200'lü ekstresinde Acinetobacter baumannii suşuna karşı en geniş zon çapı (22 mm) görülmüştür. Ayrıca ScCO2-100 ve ScCO2-200’lü ekstresi Acinetobacter baumannii suşuna karşı konsantrasyona bağlı önemli antibiyofilm aktivite sergilemiştir. Sonuç: Bu bulgular, Sideritis brevidens’in çoklu ilaç direnci gösteren bakteriyel enfeksiyonlara doğal bir antimikrobiyal ve antibiyofilm ajan kaynağı olarak önemli potansiyelini vurgulamaktadır. ScCO2 ekstraksiyonu, biyoaktif fitokimyasalların seçici olarak elde edilmesinde önemli avantajlar sunmaktadır.

Anahtar Kelimeler

Sideritis brevidens , süperkritik CO2 ekstraksiyon , soxhlet ekstraksiyon , antimikrobiyal ve antibiyofilm

Kaynakça

• Pacheco RL, Latorraca CD. OC, Dos Santos APP, et al. Efficacy and safety of home-based intravenous antibiotic therapy among adults: A systematic review. International Journal of Antimicrobial Agents. 2022;59(4):106555. doi:10.1016/j.ijantimicag.2022.106555.
• Adodo E, Ahamioje D, Ibitoye YI, Omoruyi IM. Antibiogramic efficacy of Cymbopogan citratus, Ocimum gratissimum, and Kalanchoe pinnata methanolic and aqueous extracts on colistin-resistant non-clinical isolates of Gram-negative bacteria. UMYU Scientifica. 2024;3(2):102-113. doi:10.56919/usci.2432.011
• Thompson T. The staggering death toll of drug-resistant bacteria. Nature. 2022. doi:10.1038/d41586-022-00228-x.
• Tong SY, Davis JS, Eichenberger E, Holland,TL, Fowler Jr VG. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management. Clinical Microbiology Reviews. 2015;28(3):603-661. doi:10.1128/CMR.00134-14.
• Pompilio A, Scribano D, Sarshar M, Di Bonaventura G, Palamara AT, Ambrosi C. Gram-negative bacteria holding together in a biofilm: the Acinetobacter baumannii way. Microorganisms. 2021; 9:1353. doi: 10.3390/microorganisms9071353.
• González-Burgos E, Carretero ME, Gómez-Serranillos MP. Sideritis spp. Uses, chemical composition and pharmacological activities A review. Journal of Ethnopharmacolog. 2011;135(2): 209-225. doi:10.1016/j.jep.2011.03.014.
• Güvenç A, Duman H. Morphological and anatomical studies of annual taxa of Sideritis L.(Lamiaceae), with notes on chorology in Turkey. Turkish Journal of Botany. 2010;34(2): 83-104. doi:10.3906/bot-0904-2.
• Özdemir E, Alpınar K. An ethnobotanical survey of medicinal plants in western part of central Taurus Mountains: Aladaglar (Nigde–Turkey). Journal of Ethnopharmacology. 2015;166:53-65. doi:10.1016/j.jep.2015.02.052.
• Sargın SA, Akçicek E, Selvi S. An ethnobotanical study of medicinal plants used by the local people of Alaşehir (Manisa) in Turkey. Journal of Ethnopharmacology. 2013;150(3):860-874. doi: 10.1016/j.jep.2013.09.040.
• Bulut G, Haznedaroğlu MZ, Doğan A, Koyu H, Tuzlacı E. An ethnobotanical study of medicinal plants in Acipayam (Denizli-Turkey). Journal of Herbal Medicine. 2017;10:64-81. doi:10.1016/j.hermed.2017.08.001.
• Sargin SA. Ethnobotanical survey of medicinal plants in Bozyazı district of Mersin, Turkey. Journal of Ethnopharmacology. 2015;173:105-126. doi: 10.1016/j.jep.2015.07.009.
• Aneva I, Zhelev P, Kozuharova E, Danova K, Nabavi SF, Behzad S. Genus Sideritis, section Empedoclia in southeastern Europe and Turkey–studies in ethnopharmacology and recent progress of biological activities. DARU Journal of Pharmaceutical Sciences. 2019;27(1):407-421. 27:407–421. doi:10.1007/s40199-019-00261-8.
• Yıldırım M, Erşatır M, Poyraz S, Amangeldinova M, Kudrina NO, Terletskaya NV. Green extraction of plant materials using supercritical CO2: insights into methods, analysis, and bioactivity. Plants. 2024;13(16):2295. doi:10.3390/plants13162295.
• Ramluckan K, Moodley KG, Bux F. An evaluation of the efficacy of using selected solvents for the extraction of lipids from algal biomass by the soxhlet extraction method. Fuel. 2014;116:103-108. doi:10.1016/j.fuel.2013.07.118.
• Chemat F, Rombaut N, Sicaire AG, Meullemiestre A, Fabiano-Tixier AS, Abert-Vian M. Ultrasound assisted extraction of food and natural products. Mechanisms, techniques, combinations, protocols and applications. A review. Ultrasonics Sonochemistry. 2017;34:540-560. doi:10.1016/j.ultsonch.2016.06.035.
• Li X, Zhang Y, Wang S, Shi C, Wang S, Wang X, et al. Review on the potential use of natural products in overweight and obesity. Phytotherapy Research. 2022;36(5):1990-2015. doi: 10.1002/ptr.7426.
• Amangeldinova M, Ersatır M, Necip A, et al. Simultaneous quantitative screening of 53 phytochemicals from Rheum tataricum L. roots: a comparative study of supercritical CO2, subcritical ethanol, and ultrasound-assisted extraction for enhanced antioxidant, antibacterial activities, and molecular docking study. Frontiers in Plant Science. 2024;15:1513875. doi:10.3389/fpls.2024.1513875.
• Kubátová A, Lagadec AJ, Miller DJ, Hawthorne SB. Selective extraction of oxygenates from savory and peppermint using subcritical water. Flavour and Fragrance Journal. 2001;16(1):64-73. doi:10.1002/1099-1026(200101/02)16:1<64::AID-FFJ949>3.0.CO;2-D.
• Amangeldinova M, Ersatır M, Necip A, et al. Green extraction strategies and bioactivity of Rheum cordatum Losinsk: Antioxidant, antimicrobial, and molecular docking insights. Plants. 2025;14(7):1071. doi:10.3390/plants14071071.
• Yildirim M, Necip A, Cimentepe M, Emire Z, Yabalak E. Bioactive potential of Origanum munzurense kit tan & Sorger Extracts: Antioxidant, anticancer, and antibacterial activities explored through Sc-CO2 extraction and molecular docking analysis. Journal of Chromatography. B, 2025;124614. doi:10.1016/j.jchromb.2025.124614.
• Yilmaz MA. Simultaneous quantitative screening of 53 phytochemicals in 33 species of medicinal and aromatic plants: A detailed, robust and comprehensive LC–MS/MS method validation. Industrial Crops and Products, 2020;149:112347. doi:10.1016/j.indcrop.2020.112347.
• Patel, J. B. (Ed.).Performance standards for antimicrobial susceptibility testing. Clinical and laboratory standards institute. 2017.
• Amangeldinova M, Ersatır M, Küce Cevik P, et al. Bioactive potential of Rheum cordatum Losinsk. Leaf Extracts: Phytochemical insights from supercritical CO2, subcritical ethanol and ultrasound-assisted extractions. Plants. 2025;14(15):2314. doi:10.3390/plants14152314
• Hudzicki J. Kirby-Bauer Disk Diffusion Susceptibility Test Protocol. American Society for Microbiology. 2009;15(1):1-23.
• Yildirim M, Cimentepe M, Dogan K, et al. Thymoquinone-loaded pHEMA cryogel membranes for superior control of Staphylococcus aureus infections. Russian Journal of General Chemistry. 2024;94(11):3079-3089. doi:10.1134/S1070363224110318.
• Yildirim M, Yasar E, Necip A, Cimentepe M, Demirbağ B, Kilic A. Facile synthesis and spectral analysis of the bioactive spiroborate compounds as a novel therapeutic agent for computational insights, biological evaluation, and applications. Journal of Organometallic Chemistry, 2025;1027, 123510. doi:10.1016/j.jorganchem.2025.123510.
• Yoshida H, Kawai F, Obayashi E, et al. Crystal structures of penicillin-binding protein 3 (PBP3) from methicillin-resistant Staphylococcus aureus in the Apo and cefotaxime‐bound forms. Journal of Molecular Biolog. 2012;423(3):351-364.28. doi:10.1016/j.jmb.2012.07.012.
• Derin O, Şeker F, Aksoy N, Yigit P, Yılmaz M, Mert A. The effect of antimicrobial policy implementation on carbapenem resistance: A university hospital experience. Clinical and Experimental Health Sciences. 2022;12(3):678-682. doi:10.33808/clinexphealthsci.1010571.
• Çarıkçı S, Kılıç T, Gören AC, Dirmenci T, Alim Toraman GÖ, Topçu G. Chemical profile of the Anatolian Sideritis species with bioactivity studies. Pharmaceutical Biology. 2023;61(1):1484-1511. doi:10.1080/13880209.2023.2280253.
• Lytra K, Tomou EM, Chrysargyris A, Drouza C, Skaltsa H, Tzortzakis N. Traditionally used Sideritis cypria Post.: Phytochemistry, nutritional content, bioactive compounds of cultivated populations. Frontiers in Pharmacology. 2020;11:650. doi:10.3389/fphar.2020.00650.
• Yildirim M, Kilic A, Cimentepe M, Necip A, Turedi S. Synthesis of bioactive quercetin-boronate esters as a novel biological agent: Enzyme inhibition, anti-microbial properties, computational insights and anti-cancer activity. Journal of Molecular Structure. 2025;1321:140216. doi:10.1016/j.molstruc.2024.140216.
• Tian Q, Wei S, Su H, et al. Bactericidal activity of gallic acid against multi-drug resistance Escherichia coli. Microbial Pathogenesis. 2022;173:105824. doi:10.1016/j.micpath.2022.105824
• Chen K, Peng C, Chi F, Yu C, Yang Q, Li Z. Antibacterial and antibiofilm activities of chlorogenic acid against Yersinia enterocolitica. Frontiers in Microbiology. 2022;13:885092. doi:10.3389/fmicb.2022.885092.
• Koutsaviti A, Bazos I, Milenković M, Pavlović-Drobac M, Tzakou O. Antimicrobial activity and essential oil composition of five Sideritis taxa of Empedoclia and Hesiodia sect. from Greece. Records of Natural Products. 2013;7(1):6-14.
• Aligiannis N, Kalpoutzakis E, Chinou IB, Mitakou S, Gikas E, Tsarbopoulos A. Composition and antimicrobial activity of the essential oils of five taxa of Sideritis from Greece. Journal of Agricultural and Food Chemistry. 2001;49(2):811-815. doi:10.1021/jf001018w.
• Gülcan Z, Saltan N, İşcan G, Kürkçüoğlu M, Köse YB. Antimicrobial and antioxidant activities of Sideritis lanata L. extracts. European Journal of Life Sciences. 2022;1(2):63-79. doi:10.55971/EJLS.1181461.
• Kostadinova E, Alipieva K, Stefova M, et al. Influence of cultivation on the chemical composition and antimicrobial activity of Sideritis spp. Pharmacognosy Magazine. 2008;4(14):102.
• Tadić V, Bojović, D., Arsić, I, et al. Chemical and antimicrobial evaluation of supercritical and conventional Sideritis scardica Griseb., Lamiaceae extracts. Molecules. 2012;17(3):2683-2703. doi: 10.3390/molecules17032683.
• Jamal M, Ahmad W, Andleeb S, et al. Bacterial biofilm and associated infections. Journal of the Chinese Medical Association.2018;81(1):7-11. doi:10.1016/j.jcma.2017.07.012
• Pınarlı, Y.T, Canlı, K. Determination of antibiofilm activity of Sideritis trojana. International Journal of Academic and Applied Research (IJAAR). 2021;5(11):135-138.