Evaluation of Antimicrobial and Antibiofilm Effects of Kojic Acid Derivatives against Bacteria and Fungi

Öz

Objective: In this study, it was aimed to investigate the antibacterial and antifungal activities of kojic acid derivatives against Gram-negative bacteria (Pseudomonas aeruginosa and Escherichia coli), Gram-positive bacteria (Enterococcus faecalis and Staphylococcus aureus) and fungal strain (Candida albicans). Material and Methods: Antimicrobial activity profiles of the compounds were investigated by microdilution and disk diffusion method. Antibiofilm activities were investigated by spectrophotometric microplate method including crystal violet staining. Moreover, the impact of the compounds on both biofilm and cells was also clarified via scanning electron microscopy (SEM). Results: Minimum inhibitory concentration values were found between 16 μg/mL to 1024 μg/mL. The most potent substance, 3b bearing 3,4- dichlorobenzylpiperazine moiety, has been shown to have inhibitory effects at 16-64 μg/mL concentrations. It was determined that 3a and 3b had stronger inhibitory effects than the other substances against C. albicans. In addition, antibacterial activity was discovered to be greater against Gram-positive strains than Gram-negative. It was also shown by SEM that compound 3b caused significant deformation on the cell wall and membrane of E. faecalis strain and inhibited the biofilm structures of S. aureus and E. coli strains. Conclusion: Kojic acid derivatives were found to inhibit the growth of both bacterial and fungal pathogens at low concentrations. It was also shown that kojic acid derivatives were able to decrease biofilm formation. It is thought that, this will be a favourable scenario for novel therapeutic candidates since more novel compounds will be created, and the inhibitory mechanism will be explored in various research including phenotypic and genotypic experiments.

Anahtar Kelimeler

• Kojic acid derivatives
• antimicrobial activity
• antibiofilm activity
• scanning electron microscopy

Kojik Asit Türevlerinin Bakteri ve Mantarlara Karşı Antimikrobiyal ve Antibiyofilm Etkilerinin Değerlendirilmesi

Öz

Amaç: Bu çalışmada kojik asit türevlerinin Gram-negatif bakterilere (Pseudomonas aeruginosa ve Escherichia coli), Gram-pozitif bakterilere (Enterococcus faecalis ve Staphylococcus aureus) ve mantar kökenine (Candida albicans) karşı antibakteriyel ve antifungal aktivitelerinin araştırılması amaçlanmıştır. Gereç ve Yöntem: Bileşiklerin antimikrobiyal aktivite profilleri mikrodilüsyon ve disk difüzyon yöntemleri ile araştırıldı. Antibiyofilm aktiviteler kristal viyole ile boyamayı içeren spektrofotometrik mikroplaka yöntemi ile araştırıldı. Ayrıca bileşiklerin hem biyofilm hem de hücreler üzerindeki etkisi taramalı elektron mikroskobu (SEM) ile açıklığa kavuşturuldu. Bulgular: Minimum inhibitör konsantrasyon değerleri 16 μg/mL ile 1024 μg/mL arasında bulundu. En güçlü madde olan, 3,4-diklorobenzilpiperazin kısmını taşıyan 3b'nin 16-64 μg/mL konsantrasyonlarda inhibitör etkiye sahip olduğu gösterildi. 3a ve 3b'nin C. albicans'a karşı diğer bileşiklere göre daha güçlü inhibitör etkileri olduğu saptandı. Ek olarak, antibakteriyel aktivitenin Gram-pozitif suşlara karşı, Gram-negatif suşlara göre daha fazla olduğu belirlendi. Ayrıca bileşik 3b'nin, SEM ile E. faecalis suşunun hücre duvarı ve membranında önemli deformasyona neden olduğu ve S. aureus ve E. coli suşlarının biyofilm yapılarını inhibe ettiği gösterildi. Sonuç: Kojik asit türevlerinin düşük konsantrasyonlarda hem bakteriyel hem de fungal patojenlerin büyümesini engellediği belirlenmiştir. Ayrıca kojik asit türevlerinin biyofilm oluşumunu azaltabildiği gösterilmiştir. Bu, daha fazla yeni bileşik yaratılacağı ve engelleyici mekanizmanın fenotipik ve genotipik deneyler de dahil olmak üzere çeşitli araştırmalarda keşfedileceği için yeni terapötik adaylar için olumlu bir senaryo olacaktır

Anahtar Kelimeler

• Kojik asit türevleri
• antimikrobiyal aktivite
• antibiyofilm aktivite
• taramalı elektron mikroskobu

Kaynakça

1. Burdock GA, Soni MG, Carabin IG. Evaluation of health aspects of kojic acid in food. Regul Toxicol Pharmacol [Internet]. 2001 [cited 2023 Dec 24];33(1):80–101.
2. Costerton JW, Stewart PS, Greenberg EP. Bacterial biofilms: A common cause of persistent infections. Science [Internet]. 1999 May 21 [cited 2023 Dec 24];284(5418):1318–22. DOI: 10.1126/science.284.5418.1318
3. Döşler S, Mataracı E, Başpınar-Küçük H, Yusufoğlu A. Antibacterial and anti-biofilm activities of new chiral and racemic 1,3-Dioxolanes. J Fac Pharm Istanbul. 2015;45(1):19–28.
4. Maurya VK, et al. Synthesis and evaluation of Zn(II) dithiocarbamate complexes as potential antibacterial, antibiofilm, and antitumor agents. J Coord Chem [Internet]. 2019 Nov 2 [cited 2023 Dec 23];72(19–21):3338–58. DOI: 10.1080/00958972.2019.1693041
5. Aytemir MD, Çaliş Ü, Özalp M. Synthesis and evaluation of anticonvulsant and antimicrobial activities of 3-hydroxy-6-methyl-2-substituted 4H-pyran-4-one derivatives. Arch Pharm (Weinheim). 2004;337(5):281–8.
6. Karakaya G, Aytemir MD, Özçelik B, Çaliş Ü. Design, synthesis and in vivo/in vitro screening of novel chlorokojic acid derivatives. J Enzyme Inhib Med Chem. 2013;28(3):627–38.
7. Aytemir MD, Özçelik B. A study of cytotoxicity of novel chlorokojic acid derivatives with their antimicrobial and antiviral activities. Eur J Med Chem [Internet]. 2010;45(9):4089–95. DOI: 10.1016/j.ejmech.2010.05.069
8. Karakaya G, Ercan A, Öncül S, Aytemir MD. Kojic acid derivatives as potential anticancer agents: Synthesis and cytotoxic evaluation on A375 human malignant melanoma cells. J Res Pharm. 2019;23(4):596–607.

9. Karakaya G, Ercan A, Oncul S, Aytemir MD. Synthesis and cytotoxic evaluation of kojic acid derivatives with inhibitory activity on melanogenesis in human melanoma cells. Anticancer Agents Med Chem. 2018;18(15):2137–48.
10. Ercan A, Oncul S, Karakaya G, Aytemir M. An allomaltol derivative triggers distinct death pathways in luminal A and triple-negative breast cancer subtypes. Bioorg Chem [Internet]. 2020;105(July):104403. DOI: 10.1016/j.bioorg.2020.104403
11. Aytemir M, et al. United States Patent, Patent No.: US 9,975,884 B2. Kojic acid - derived Mannich bases with biological effect. Date of Patent: May 22, 2018.
12. Karakaya G, et al. Synthesis, molecular docking and tyrosinase inhibition of kojic acid derivatives. Bioorg Chem [Internet]. 2019;88(January):102950. DOI: 10.1016/j.bioorg.2019.102950
13. Karakaya G, et al. Synthesis and modeling of novel hydroxypyrone derivatives as antidermatophytic agents. J Heterocycl Chem. 2022;59(10):1801–12.
14. Stoykova B, et al. Synthesis and antimicrobial activity of novel kojyl carbamates. Bulg Chem Commun. 2020;52:23-9.
15. Khodabandlou S, Saraei M. Synthesis of novel isoxazole derivatives bearing kojic acid moiety and evaluation of their antimicrobial activity. Chem Heterocycl Compd. 2021;57(7–8):823–7.
16. Azhar SNAS, et al. In vitro kinetic release study, antimicrobial activity and in vivo toxicity of kojic acid ester-based nanoemulsion. RSC Adv. 2020;10(71):43894–903.
17. Ezzat H, et al. Enhanced anti-bacterial effect of kojic acid using gelatinized core liposomes. J Drug Deliv Sci Technol [Internet]. 2021;64(March):102625.
18. Chaudhary J, et al. Application of kojic acid immobilised magnetic and chitosan tri-polyphosphate nanoparticles as antibacterial agents. IET Nanobiotechnology. 2015;9(6):375–80.
19. Hussein-Al-Ali SH, et al. Novel kojic acid-polymer-based magnetic nanocomposites for medical applications. Int J Nanomedicine. 2014;9(1):351–62.
20. Liu X, et al. Development of kojic acid and chitosan composite biodegradable films. Int J Biol Macromol [Internet]. 2020;144:483–90. DOI: 10.1016/j.carbpol.2021.117778
21. Aytemir MD, Özçelik B, Karakaya G. Evaluation of chlorokojic acid derivatives against dermatophytes. Bioorg Med Chem Lett. 2013;23(12):3646–9.
22. Kotmakçı M, et al. Characterization and antimicrobial activity of hazelnut oil microemulsion loaded with mitomycin C. Lat Am J Pharm. 2015;34:529–36.
23. Başpınar Y, et al. Antimicrobial activity of phytosphingosine nanoemulsions. Celal Bayar Univ Sci J. 2018;14(2):223–8.
24. Öztürk İ, et al. In vitro effects of antibiofilm agents and antibiotics on coagulase-negative staphylococci. J Res Pharm. 2020;24:821–32.
25. Kahraman Vatansever S, et al. Effect of Bdellovibrio bacteriovorus on pathogens and biofilms. Indian J Microbiol [Internet]. 2023;63(1):139–45. DOI: 10.1007/s12088-023-01071-y
26. Weber K, et al. Comparison of SEM and VPSEM imaging with respect to Streptococcus mutans biofilm. FEMS Microbiol Lett [Internet]. 2014;350(2):175–9. DOI: 10.1111/1574-6968.12334
27. Golding CG, et al. The scanning electron microscope in microbiology. Sci Rep [Internet]. 2016;6(1):1–8. DOI: 10.1038/srep26516
28. Aslam B, et al. Antibiotic resistance: a global crisis. Infect Drug Resist [Internet]. 2018;11:1645–58.
29. Pulingam T, et al. Antimicrobial resistance: prevalence and strategies. Eur J Pharm Sci. 2022;170:106103.
30. Mancuso G, et al. Bacterial antibiotic resistance: the most critical pathogens. Pathogens [Internet]. 2021;10(10):1310. DOI: 10.3390/pathogens12010116
31. Jorge P, et al. New trends in peptide-based anti-biofilm strategies. Biofouling [Internet]. 2012;28(10):1033–61. DOI: 10.1080/08927014.2012.728210
32. Wu Y, et al. Evaluation of kojic acid antibacterial and anti-biofilm properties. Food Sci Technol Int [Internet]. 2019;25(1):3–15. DOI: 10.1177/1082013218793075
33. Li H, et al. Kojic acid combating Acinetobacter baumannii biofilm. Microbiol Res [Internet]. 2022;254:126911.
34. Pantanella F, et al. Analytical techniques to study microbial biofilm. Ann Ig [Internet]. 2013;25(1):31–42. DOI: 10.7416/ai.2013.1904
35. Stiefel P, et al. Is biofilm removal properly assessed? Appl Microbiol Biotechnol [Internet]. 2016;100(9):4135–45. DOI: 10.1007/s00253-016-7396-9