Investigation of İn Vitro Antimicrobial Activity of Pomegranate (Punica Granatum L.) Fruit Peel Extracts

Öz

In recent years, the use of plant-derived products and dietary supplements has significantly increased, leading to a rise in scientific research on their biological effects. Among these, the antimicrobial properties of plants have received particular attention. Plants naturally contain a wide variety of secondary metabolites, such as tannins, terpenoids, alkaloids, and flavonoids, which are known to possess antimicrobial activity. Punica granatum L. (pomegranate) is especially rich in tannins and has been reported to exhibit notable antimicrobial potential. In this study, the antimicrobial activities of water, ethanol, dichloromethane, and ethyl acetate extracts of Punica granatum fruit peels were evaluated against five standard strains: Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Candida albicans ATCC 10231. The fruit peels were dried, ground into powder, and subjected to direct solvent extraction. The minimum inhibitory concentration values of each extract were determined using the microdilution method. The extracts exhibited the highest antimicrobial activity against Staphylococcus aureus, with the ethanol and water extracts being the most effective. Ethanol, water, and ethyl acetate extracts also showed inhibitory effects on Enterococcus faecalis. However, no antimicrobial activity was observed against Pseudomonas aeruginosa, Escherichia coli, or Candida albicans. Among all extracts, the ethanol extract demonstrated the strongest antimicrobial effect, while the dichloromethane extract showed no antimicrobial activity under the tested conditions. These findings suggest that pomegranate peels, often discarded as waste, could serve as a natural preservative in the food industry and as a potential natural alternative for addressing the growing challenge of antibiotic resistance in pharmaceutical applications.

Anahtar Kelimeler

• Antimicrobial activity
• phenolic
• microdilution method
• pomegranate peel extract
• Punica granatum L.

Nar (Punica Granatum L.) Meyve Kabuğu Ekstraktlarının İn Vitro Antimikrobiyal Etkinliğinin Araştırılması

Öz

Bitkilerden elde edilen ürünlerin ve gıda takviyelerinin kullanımı son yıllarda artmıştır ve bunun üzerine pek çok çalışma yapılmıştır. Bu çalışmalar arasında bitkilerin antimikrobiyal aktivite göstermesine yönelik çalışmalar da vardır. Bitkiler; antimikrobiyal özelliklere sahip tanenler, terpenoidler, alkaloidler ve flavonoidler gibi çok çeşitli ikincil metabolitler açısından zengindir. Punica granatum L. (Nar) antimikrobiyal aktiviteye sahip tanenler açısından zengin bir bitkidir. Çalışmamızda Punica granatum L. meyve kabuklarından elde edilen su, etanol, diklorometan ve etil asetat ekstraktlarının Enterococcus faecalis ATCC 29212, Staphylococcus aureus ATCC 29213, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853 bakteri suşlarına ve Candida albicans ATCC 10231 mantar suşuna karşı antimikrobiyal aktivitesi araştırılmıştır. Nar meyve kabukları kurutulup toz haline getirildikten sonra farklı çözücüler kullanılarak direkt özütleme yöntemi ile ekstraktlar hazırlanmıştır. Ekstraktların minimum inhibitör konsantrasyon değerleri mikrodilüsyon yöntemi ile belirlenmiştir. Ekstraktlar en yüksek antimikrobiyal aktiviteyi Staphylococcus aureus suşuna karşı göstermiştir; bu suşa karşı en etkili ekstraktlar su ve etanol ekstraktlarıdır. Enterococcus faecalis suşuna karşı ise etanol, su ve etil asetat ekstraktları antimikrobiyal etki göstermiştir. Gram negatif özellikteki Pseudomonas aeruginosa ve Escherichia coli suşları ile Candida albicans suşunda antimikrobiyal aktiviteye rastlanmamıştır. Çalışmada kullanılan ekstraktlar arasında en yüksek antimikrobiyal aktiviteyi etanol ekstraktı gösterirken, diklorometan ekstraktı çalışmada kullanılan koşullar altında test edilen suşlar üzerinde antimikrobiyal aktivite göstermemiştir. Bu bulgular, atık olarak değerlendirilen nar meyvesi kabuğunun gıda endüstrisinde doğal koruyucu olarak ve ilaç endüstrisinde artan antibiyotik direncine karşı doğal bir alternatif olarak kullanılma potansiyeline sahip olduğunu düşündürmektedir.

Anahtar Kelimeler

• Antimikrobiyal aktivite
• fenolik
• mikrodilüsyon yöntemi
• nar kabuğu ekstraktı
• Punica granatum L.

Kaynakça

1. Pinheiro AJMCR, Gonçalves JS, Dourado ÁWA, et al. Punica granatum L. leaf extract attenuates lung inflammation in mice with acute lung injury. J Immunol Res. 2018;2018:6879183. doi:10.1155/2018/6879183.
2. Pinheiro AJMCR, Mendes ARS, Neves MDFJ, et al. Galloyl hexahydroxydiphenoyl (HHDP) glucose isolated from Punica granatum L. leaves protects against lipopolysaccharide (LPS) induced acute lung injury in BALB/c mice. Front Immunol. 2019;10:1978. doi:10.3389/fimmu.2019.01978.
3. Angelini P. Plant-derived antimicrobials and their crucial role in combating antimicrobial resistance. Antibiotics (Basel). 2024;13(8):746. doi:10.3390/antibiotics13080746.
4. El-Saadony MT, Saad AM, Mohammed DM, et al. Medicinal plants: bioactive compounds, biological activities, combating multidrug-resistant microorganisms, and human health benefits:a comprehensive review. Front Immunol. 2025;16:1491777. doi:10.3389/fimmu.2025.1491777.
5. Xu S, Kang A, Tian Y, et al. Plant flavonoids with antimicrobial activity against methicillin-resistant Staphylococcus aureus (MRSA). ACS Infect Dis. 2024;10(9):3086-3097. doi:10.1021/acsinfecdis.4c00292.
6. Pagliarulo C, De Vito V, Picariello G, et al. Inhibitory effect of pomegranate (Punica granatum L.) polyphenol extracts on the bacterial growth and survival of clinical isolates of pathogenic Staphylococcus aureus and Escherichia coli. Food Chem. 2016;190:824-831. doi:10.1016/j.foodchem.2015.06.028.
7. Fawole OA, Makunga NP, Opara UL. Antibacterial, antioxidant and tyrosinase-inhibition activities of pomegranate fruit peel methanolic extract. BMC Complement Altern Med. 2012;12:200. doi:10.1186/1472-6882-12-200.
8. Sharayei P, Azarpazhooh E, Ramaswamy HS. Effect of microencapsulation on antioxidant and antifungal properties of aqueous extract of pomegranate peel. J Food Sci Technol. 2020;57(2):723-733. doi:10.1007/s13197-019-04105-w.

9. Uluman E. Nar (Punica granatum L.) kabuk ekstraktının mitomycin C genotoksisitesi üzerine olası antigenotoksik etkilerinin in vivo olarak araştırılması. Yüksek lisans tezi. Kafkas Üniversitesi Fen Bilimleri Enstitüsü, Kars; 2019.
10. Gündoğdu M, Yılmaz H, Canan İ. Nar (Punica granatum L.) çeşit ve genotiplerin fizikokimyasal karakterizasyonu. Uluslararası Tarım ve Yaban Hayatı Bilimleri Dergisi. 2015;2:57-65.
11. Demir T, Akpınar Ö, Kara H, Güngör H. Nar (Punica granatum L.) kabuğunun in vitro antidiyabetik, antienflamatuvar, sitotoksik, antioksidan ve antimikrobiyal aktivitesi. Akademik Gıda. 2019;17(1):61-71. doi:10.24323/akademik-gida.544647.
12. Brighenti V, Iseppi R, Pinzi L, et al. Antifungal activity and DNA topoisomerase inhibition of hydrolysable tannins from Punica granatum L. Int J Mol Sci. 2021;22(8):4175. doi:10.3390/ijms22084175.
13. Abdollahzadeh Sh, Mashouf R, Mortazavi H, Moghaddam M, Roozbahani N, Vahedi M. Antibacterial and antifungal activities of Punica granatum peel extracts against oral pathogens. J Dent (Tehran). 2011;8(1):1-6.
14. Emer Z, Pakyürek M. Narın (Punica granatum L.) tıbbi önemi. Uluslararası Tarım ve Kırsal Kalkınma Kongresi; 10-12 Haziran 2019, Siirt, Türkiye.
15. Dahham SS, Ali MN, Tabassum H, Khan M. Studies on antibacterial and antifungal activity of pomegranate (Punica granatum L.). Am Eurasian J Agric Environ Sci. 2010;9:273-281.
16. Parfitt J, Barthel M, Macnaughton S. Food waste within food supply chains: quantification and potential for change to 2050. Philos Trans R Soc Lond B Biol Sci. 2010;365(1554):3065-3081. doi:10.1098/rstb.2010.0126.
17. Roselli V, Pugliese G, Leuci R, et al. Green methods to recover bioactive compounds from food industry waste: a sustainable practice from the perspective of the circular economy. Molecules. 2024;29(11):2682. doi:10.3390/molecules29112682.
18. Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing. M100. 32nd ed. Wayne, PA: CLSI; 2022. https://clsi.org/standards/products/microbiology/documents/m100/. Accessed April 06, 2026.
19. European Committee on Antimicrobial Susceptibility Testing (EUCAST). Breakpoint tables for interpretation of MICs and zone diameters. Version 15; Växjö (Sweden): 2025. https://www.eucast.org/clinical_breakpoints. Accessed April 06, 2026.
20. Ghafoor K, Al-Juhaimi FY, Babiker EE. Phenolic compounds, antioxidant and antibacterial properties of peel and pulp of avocado (Persea americana). J Food Biochem. 2011;35(2):574-585.
21. Mendonça AMS, Monteiro CdA, Moraes-Neto RN, et al. Ethyl acetate fraction of Punica granatum and its galloyl-HHDP-glucose compound, alone or in combination with fluconazole, have antifungal and antivirulence properties against Candida spp. Antibiotics. 2022;11(2):265. doi:10.3390/antibiotics11020265.
22. Pereira JV, Pereira MSV, Sampaio FC, et al. In vitro antimicrobial and anti-adherence effect of Punica granatum Linn. extracts upon dental biofilm microorganisms. Braz J Pharmacogn. 2006;16(1):88-93. doi:10.1590/S0102-695X2006000100016.
23. Xu Y, Shi C, Wu Q, et al. Antimicrobial activity of punicalagin against Staphylococcus aureus and its effect on biofilm formation. Foodborne Pathog Dis. 2017;14(5):282-287. doi:10.1089/fpd.2016.2226.
24. Ahuja S, Dodwad V, Kukreja BJ, Mehra P, Kukreja P. A comparative evaluation of efficacy of Punica granatum and chlorhexidine on plaque and gingivitis. J Int Clin Dent Res Organ. 2011;3:29-32. doi:10.4103/2231-0754.115776.
25. Bhadbhade SJ, Acharya AB, Rodrigues SV, Thakur SL. The antiplaque efficacy of pomegranate mouthrinse. Quintessence Int. 2011;42(1):29-36.
26. Saquib SA, AlQahtani NA, Ahmad I, et al. Synergistic antibacterial activity of herbal extracts with antibiotics on bacteria responsible for periodontitis. J Infect Dev Ctries. 2021;15(11):1685-1693. doi:10.3855/JIDC.14904.
27. Salan AS, Usta EFN, Kuyucuklu-Kazan G, Ökten S. Papatya suyu toniğinin akne tedavisinde kullanılan antibiyotiklerin etkinliği üzerine etkisi. Int J Pure Appl Sci. 2026;12(1):54-69. doi:10.29132/ijpas.1800866.
28. Hayouni EA, Miled K, Boubaker S, et al. Hydroalcoholic extract based ointment from Punica granatum L. peels with enhanced in vivo healing potential on dermal wounds. Phytomedicine. 2011;18(11):976-984. doi: 10.1016/j.phymed.2011.02.011.
29. Gullon B, Pintado ME, Pérez Álvarez JA, Viuda Martos M. Assessment of polyphenolic profile and antibacterial activity of pomegranate peel (Punica granatum) flour obtained from co-product of juice extraction. Food Cont. 2016;59(1):94-98 doi:10.1016/j.foodcont.2015.05.025.
30. Kiran S, Tariq A, Iqbal S, et al. Punicalagin, a pomegranate polyphenol sensitizes the activity of antibiotics against three MDR pathogens of the Enterobacteriaceae. BMC Complement Med Ther. 2024;24(1):93. doi:10.1186/s12906-024-04376-7.
31. Prashanth D, Asha MK, Amit A. Antibacterial activity of Punica granatum. Fitoterapia. 2001;72(2):171-173. doi: 10.1016/s0367-326x(00)00270-7.
32. Alexandre EMC, Silva S, Santos SAO, et al. Antimicrobial activity of pomegranate peel extracts performed by high pressure and enzymatic assisted extraction. Food Res Int. 2019;115:167-176. doi:10.1016/j.foodres.2018.08.044.
33. Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev. 1999;12(4):564-582.
34. Konca T. Punica granatum L. bitkisinin antibakteriyel etkisinin araştırılması. Yüksek lisans tezi. Sakarya Üniversitesi Fen Bilimleri Enstitüsü; 2012, Sakarya.
35. Endo EH, Cortez DAG, Ueda-Nakamura T, Nakamura CV, Dias Filho BP. Potent antifungal activity of extracts and pure compound isolated from pomegranate peels and synergism with fluconazole against Candida albicans. Res Microbiol. 2010;161(7):534-540. doi:10.1016/j.resmic.2010.05.002.
36. Magangana TP, Makunga NP, Fawole OA, Opara UL. Effect of solvent extraction and blanching pre-treatment on phytochemical, antioxidant properties, enzyme inactivation and antibacterial activities of ‘Wonderful’ pomegranate peel extracts. Processes. 2021;9(6):1012. doi:10.3390/pr9061012.
37. Mun SH, Kang OH, Kong R, et al. Punicalagin suppresses methicillin resistance of Staphylococcus aureus to oxacillin. J Pharmacol Sci. 2018;137(4):317-323. doi:10.1016/j.jphs.2017.10.008.
38. Tariq A, Sana M, Shaheen A, et al. Restraining the multidrug efflux transporter STY4874 of Salmonella Typhi by reserpine and plant extracts. Lett Appl Microbiol. 2019;69(3):161-167. doi:10.1111/lam.13196.
39. Song W, Wang L, Jin M, et al. Punicalagin, an inhibitor of Sortase A, is a promising therapeutic drug to combat methicillin-resistant Staphylococcus aureus infections. Antimicrob Agents Chemother. 2022;66(6):e00224-22. doi:10.1128/aac.00224-22.
40. Salan AS, Ökten S. Uzun süreli kullanılan ilaçların antimikrobiyal etkisinin ve antimikrobiyal ajanlar ile in vitro etkileşiminin araştırılması. Mikrobiyol Bul. 2025;59(3):269-291. doi:10.5578/mb.202503165.
41. Ökten S, Salan AS, Kuyucuklu G, Kaynak Onurdağ F. Acinetobacter spp. izolatlarında dışa atım pompası (DAP) inhibitörlerinin meropenemin etkinliği üzerine etkisi. J Fac Pharm Ankara. 2022;46(1):35-47. doi:10.33483/jfpau.995682.
42. Salan AS, Ökten S. The antimicrobial effect of N-acetylcysteine and its interaction with antibiotics against Acinetobacter baumannii isolates. J Fac Pharm Ankara. 2024;48(2):396-405. doi:10.33483/jfpau.1358483.
43. Li G, Xu Y, Pan L, Xia X. Punicalagin damages the membrane of Salmonella typhimurium. J Food Prot. 2020;83(12):2102-2106. doi: 10.4315/JFP-20-173.
44. Li G, Yan C, Xu Y, et al. Punicalagin inhibits Salmonella virulence factors and has anti-quorum-sensing potential. Appl Environ Microbiol. 2014;80(19):6204-6211. doi:10.1128/AEM.01458-14.
45. Safdar M, Naqvi SA, Anjum F, et al. Microbial biofilm inhibition, antioxidants, and chemical fingerprints of Afghani pomegranate peel extract documented by gas chromatography–mass spectrometry and Fourier transformation infrared. J Food Process Preserv. 2021;45(7):e15657. doi:10.1111/jfpp.15657.