Evaluation of Cytotoxicity and Antimicrobial Activity of Flavipin Against Acinetobacter baumannii, Methicillin-Resistant Staphylococcus aureus, and Candida albicans



  • Mohammed J. Qasim Department of Microbiology, College of Medicine, Al-Nahrain University, Baghdad- Iraq
  • Azhar A.F. Al-Attraqchi Department of Microbiology, College of Medicine, Al-Nahrain University, Baghdad- Iraq
  • Gazi M. Aziz Department of Biotechnology, College of Science, Baghdad University, Baghdad-Iraq


Methicillin-resistant Staphylococcus aureus, IC50, Flavipin, Candida albicans, Aspergillus flavus, Antimicrobial activity, Acinetobacter baumannii


Flavipin, a metabolite produced by Aspergillus species is a potent antibiotic, antifungal, anticancer, and nematocidal agent. The present study was conducted to investigate the cytotoxicity and antimicrobial activity of flavipin produced by Aspergillus flavus. Eighty isolates of Aspergillus species were screened for the presence of flavipin. Macroscopic, microscopic, and molecular characterizations were performed to identify the Aspergillus species. Flavipin extract was prepared, fractionated, and the concentration was determined in the selected isolates. The antimicrobial activity of flavipin was tested against Candida albicans, Acinetobacter baumannii, and methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and minimum fungicidal concentration (MFC) were determined. A cytotoxicity test of flavipin was conducted on MCF-7 and WRL-68 cell lines. The results of the antibacterial activity of flavipin against MRSA and A. baumannii showed a wide zone of inhibition of 35 and 30 mm, respectively. Flavipin exhibited the highest antifungal activity against C. albicans, with inhibition zones of 44 mm in diameter. The MICs of flavipin for both A. baumannii and MRSA were 31.2 μg/mL, while the MIC of flavipin for C. albicans was 62.5 μg/mL. The MBC values of flavipin for MRSA and A. baumannii were 62.5 and 125 μg/mL, respectively, while the MFC of flavipin against C. albicans was 125 μg/mL. The IC50 of flavipin was 52.34 μg/mL for the cancer cell line MCF-7 and 298.1 μg/mL for the normal cell line WRL-68. The findings of the study revealed that flavipin produced by Aspergillus flavus has antimicrobial and cytotoxic properties. 


Deshmukh S, Dufossé L, Chhipa H, Saxena S, Mahajan G, Gupta M. Fungal Endophytes: A potential source of antibacterial compounds. J Fungi. 2022; 8(2):164.

Coletto S, Nascimento T, Cabral A, Oliveira C. Characterization of endophytes of grapevine belonging to Epicoccum nigrum complex and its interaction with grapevine trunk fungi. Rev Fac Cienc Agrar. 2018; 41(Special): 67-81.

Flewelling A, Bishop A, Johnson J, Gray C. Polyketides from an Endophytic Aspergillus fumigatus isolate inhibit the growth of Mycobacterium tuberculosis and MRSA. Nat Prod Commun. 2015; 10(10): 1661 - 1662.

Burge W, Buckley L, Sullivan J, McGrattan C, Ikawa M. Isolation and biological activity of the pigments of the mold Epicoccum nigrum. J Agric Food Chem. 1976; 24(3):555- 559.

Raistrick H, Rudman P. Studies in the biochemistry of micro-organisms. 97. Flavipin, a crystalline metabolite of Aspergillus flavipes (Bainier & Sartory) Thom & Church and Aspergillus terreus Thom. Biochem. J. 1956; 63(3):395- 406.

Brown A, Finlay R, Ward J. Antifungal compounds produced by Epicoccum purpurascens against soil-borne plant pathogenic fungi. Soil Biol Biochem. 1987; 19(6):657- 664.

Madrigal C, Melgarejo P. Mechanisms of action of the antibiotic flavipin on Monilinia laxa and Saccharomyces cerevisiae. Mycol Res. 1994; 98(8):874-878.

Hanieh H. Aryl hydrocarbon receptor-microRNA-212/132 axis in human breast cancer suppresses metastasis by targeting SOX4. Mol Cancer. 2015; 14(1):1-13.

De la Maza L, Pezzlo M, Baron E. Color atlas of diagnostic microbiology. St. Louis: Mosby; 1997. ISBN 13: 9780815106210.

Ellis D, Davis S, ALexiou H, Handke R, Bartly R. Description of medical fungi. 2nd ed. The nat lib of Aust Catal In pnb en: university of Adelaide, Australia; 2007.

Alshehri B, Palanisamy M. Evaluation of molecular identification of Aspergillus species causing fungal keratitis. Saudi J Biol Sci. 2020; 27(2):751-756.

Ye Y, Xiao Y, Ma L, Li H, Xie Z, Wang M et al. Flavipin in Chaetomium globosum CDW7, an endophytic fungus from Ginkgo biloba, contributes to antioxidant activity. Appl Microbiol Biotechnol. 2013; 97(16):7131-7139.

CLSI, Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically: Approved Standard, 9th ed, CLSI, 2012. 2022.

The European Committee on Antimicrobial Susceptibility Testing. Routine and extended internal quality control for MIC determination and disk diffusion as recommended by EUCAST. Version 12.0, 2022. http://www.eucast.org

Ohikhena F, Wintola O, Afolayan A. Evaluation of the antibacterial and antifungal properties of Phragmanthera capitata (Sprengel) Balle (Loranthaceae), a mistletoe growing on rubber tree, using the dilution techniques. Sci World J. 2017; 2017:1-8.

Gutef E. Evaluation of some antimicrobial activity of different Pleurotus ostreatus extracts in vitro and in vivo. PhD thesis. 2020; Micro Dep/ Coll of Med/AL. Nah Uni/Bag.

Jasim A, Kumar Y, Benjamin J, Masih H. Studies on antifungal properties of some plant extracts (garlic, fenugreek, and ginger) against of clinical isolate candida species. Int J Sci Eng Technol Res. 2013; 2(19):2180-2185.

Nader M, Ghanima K, Ali S, Azhar D. Antibacterial activity of ginger extracts and its essential oil on some of pathogenic bacteria. Baghdad Sci J. 2010; 7(3):1159-1165.

Foerster S, Desilvestro V, Hathaway L, Althaus C, Unemo M. A new rapid resazurin-based microdilution assay for antimicrobial susceptibility testing of Neisseria gonorrhoeae. J Antimicrob Chemother. 2017; 72(7):1961- 1968.

Elshikh M, Ahmed S, Funston S, Dunlop P, McGaw M, Marchant R et al. Resazurin-based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants. Biotechnol Lett. 2016; 38(6):1015-1019.

Al-Safar M, Hassan J, Abdulrhman T, Kashkol A. Antibacterial activity of bee venom against multidrugresistant Acinetobacter baumannii locally isolates. Int J Res Pharm Sci. 2018; 9(4), 1510-1514.

Asmerom D, Hailu G, Yimer E, Bitew H, Kahsay G. Antimicrobial evaluation of latex and TLC fractions from the leaves of Aloe adigratana reynolds. Evid-based Complement Altern Med. 2020; 2020:1-10.

Gharaghani M, Rezaei-Matehkolaei A, Zarei Mahmoudabadi A, Keikhaei B. The Frequency, antifungal susceptibility and enzymatic profiles of Candida species isolated from neutropenic patients. Jundishapur J Microbiol. 2016; 9(11): e41446

Meerloo J, Kaspers G, Cloos J. Cell sensitivity assays: the MTT assay. Cancer cell culture: Methods protoc. 2011; 731:237-245.

Awan Z, A. Fahmy U, Badr-Eldin S, Ibrahim T, Asfour H, Al-Rabia M et al. The enhanced cytotoxic and pro-apoptotic effects of optimized simvastatin-loaded emulsomes on MCF-7 breast cancer cells. Pharm. 2020; 12(7):597.

Senthilraja P, Kathiresan K. In vitro cytotoxicity MTT assay in Vero, HepG2 and MCF -7 cell lines study of Marine Yeast. J Appl Pharm Sci. 2015; 080-084.

Dudeja S, Chhokar V, Beniwal V, Badgujjar H, Chauhan R, Soni S et al. Optimization and production of antimicrobial compounds by Aspergillus flavus MTCC 13062 and its synergistic studies. Biocatal Agric Biotechnol. 2021; 35:102065.

Lloyd D. Topics in Carbocyclic Chemistry. Springer, Boston, MA, Volume One. 1st ed. 1969:90-99.

Okayo R, Andika D, Dida M, K’Otuto G, Gichimu B. Morphological and molecular characterization of toxigenic Aspergillus flavus from Groundnut Kernels in Kenya. Int J Microbiol. 2020; 2020:1-10.

Saif F, Yaseen S, Alameen A, Mane S, Undre P. Identification and characterization of Aspergillus species of fruit rot fungi using microscopy, FT-IR, Raman and UV–Vis spectroscopy. Spectrochim Acta A Mol Biomol Spectrosc. 2021; 246:119010.

Nazir N, Siddique A, Khan M, Ishaque M. Inhibitory effects of antifungal drugs on biofilm producing Aspergillus spp. recovered from drinking water system. J Bacteriol Mycol. 2020; 7(2), 1-6.

Henry T, Iwen P, Hinrichs S. Identification of Aspergillus species using internal transcribed spacer regions 1 and 2. J Clin Microbiol. 2000; 38(4):1510-1515.

Priya T, Mahendra C6, Phadnis S, Peter A. Morphological and molecular identification of Aspergillus flavus isolated from rice (Oryza sativa L.) grain samples in Karnataka and Tamil Nadu in Southern India. J Pure Appl Microbiol. 2016; 10(1): 535-545.

Nleya N, Ngoma L, Adetunji M, Mwanza M. Biodiversity of aflatoxigenic Aspergillus species in dairy feeds in Bulawayo, Zimbabwe. Front Microbiol. 2021; 11.

Madrigal C, Tadeo J, Melgarejo P. Relationship between flavipin production by Epicoccum nigrum and antagonism against Monilinia laxa. Mycol Res. 1991; 95(12):1375- 1381.

Xiao Y, Li H, Li C, Wang J, Li J, Wang M et al. Antifungal screening of endophytic fungi from Ginkgo biloba for discovery of potent anti-phytopathogenic fungicides. FEMS Microbiol Lett. 2013; 339(2):130-136.

Nitao J, Meyer S, Chitwood D, Schmidt W, Oliver J. Isolation of flavipin, a fungus compound antagonistic to plant-parasitic nematodes. Nematol. 2002; 4(1):55-63.

Madrigal C, Tadeo J, Melgarejo P. Degradation of flavipin by Monilinia laxa. Mycol Res. 1993; 97(5):634-636.

CLSI, Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically: Approved Standard, 9th ed, CLSI, 2012. 2022.

Flewelling A, Johnson J, Gray C. Isolation and bioassay screening of fungal endophytes from North Atlantic marine macroalgae. Bot Mar. 2013; 56(3):287-297.

McCulloch M, Berrue F, Haltli B, Kerr R. One-Pot Syntheses of Pseudopteroxazoles from Pseudopterosins: A Rapid route to non-natural congeners with improved antimicrobial activity. J Nat Prod. 2011; 74(10):2250-2256.

CLSI, Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically: Approved Standard, 9th ed, CLSI, 2012. 2022.

Harwoko H, Lee J, Hartmann R, Mándi A, Kurtán T, Müller W et al. Azacoccones F-H, new flavipin-derived alkaloids from an endophytic fungus Epicoccum nigrum MK214079. Fitoterapia. 2020; 146:104698.

Lee A, Cadelis M, Kim S, Swift S, Copp B, Villas-Boas S. Epipyrone A, a Broad-Spectrum Antifungal Compound Produced by Epicoccum nigrum ICMP 19927. Molecules. 2020; 25(24):5997.

Senthil Kumar V, Kumaresan S, Tamizh M, Hairul Islam M, Thirugnanasambantham K. Anticancer potential of NF-κB targeting apoptotic molecule “flavipin” isolated from endophytic Chaetomium globosum. Phytomedicine. 2019; 61:152830.

Hanieh H, Mohafez O, Hairul-Islam V, Alzahrani A, Bani Ismail M, Thirugnanasambantham K. Novel Aryl hydrocarbon receptor agonist suppresses migration and invasion of breast cancer cells. PLoS one. 2016; 11(12):e0167650.

Hashim ST, Fakhry SS, Rasoul LM, Saleh TH, Alrubaii BAL. Genotyping toxins of Clostridium perfringens strains of rabbit and other animal origins. Trop J Nat Prod Res. 2021; 5(4):613–616.

Mohsin MR, AL-Rubaii BA. Bacterial growth and antibiotic sensitivity of Proteus mirabilis treated with antiinflammatory and painkiller drugs. Biomedicine. 2023; 43(02):728-34.

Al-Musawi, AHO, Aziz, HM, Khudair, S, Saleh, TH. Molecular characterization of HBB gene mutations in betathalassemia patients of Southern Iraq. Biomedicine. 2022; 42(5):1040-1043.

Rasin KH, Algabar FA, Al-Obaidi M, Krayfa AH. Evaluation of Medical waste management during the COVID-19 in Diyala Governorate. Environ Eng Manag J. 2022; 21(12):1931–1944.

Eyad HN, Adbulateed YA, Lafi SA. Abnormal presentation of TB patients: anthropological study. Ann Trop Med Public Health. 2019; 22(VI): S186.

Hamoode RH, Alkubaisy SA, Sattar DA, Hamzah SS, Saleh TH, Al-Rubaii BAL. Detection of anti-testicular antibodies among infertile males using indirect immunofluorescent technique. Biomedicine. 2022; 42 (5):978-982.

Abdulrazaq, RA., Mahmood, WS., Alwan, B, Saleh, TH, Hashim, ST, Al-Rubaii, BAL. Biological study of protease produced by clinical isolates of Staphylococcus aureus. Res J Pharm Technol.2022; 15(12):5415-5420.

Rasoul LM, Allami RH, Alshibib AL, laftaah Al-Rubaii BA, Sale TH. Expression and cytotoxic effect of recombinant Newcastle Disease Virus (rNDV) vector expressing enhanced green fluorescent gene in JHH5 cell line. Biomedicine. 2023; 43(1):205-209.

Allami RH, Hassoon AH, Abdulateef YM, Ghani AA. Genetic Association of Angiotensin-converting enzyme 2 ACE-2 (rs2285666) Polymorphism with the Susceptibility of COVID-19 Disease in Iraqi Patients. Trop J Nat Prod Res. 2023;7(2):2346-2351

Kadhim AL-Imam MJ, AL-Rubaii BAL. The influence of some amino acids, vitamins and anti-inflammatory drugs on activity of chondroitinase produced by Proteus vulgaris caused urinary tract infection. Iraqi J Sci. 2016; 57(4A):2412-2421.

Jawad NK, Numan AT, Ahmed AG, Saleh TH, Al-Rubaii BA. IL-38 gene expression: A new player in Graves’ ophthalmopathy patients in Iraq. Biomedicine. 2023;43(1):210-215.

Shehab ZH, AL-Rubaii BAL. Effect of D-mannose on gene expression of neuraminidase produced from different clinical isolates of Pseudomonas aeruginosa. Baghdad Sci J. 2019;16(2):291–298.

Abdulla L, Ismael MK, Salih TA, Malik SN, Al-Rubaii BAL. Genotyping and evaluation of interleukin-10 and soluble HLA-G in abortion due to toxoplasmosis and HSV- 2 infections. Ann Parasitol. 2022; 68(2):385–390.

Jiad AL, Ismael MK, Muhsin SS, Al-Rubaii BAL. ND2 Gene Sequencing of Sub fertile Patients Recovered from COVID-19 in Association with Toxoplasmosis. Bionatura. 2022; 7(3):45.

Rasoul LM, Nsaif MM, Al-Tameemi MT, Al-Rubaii BA. Estimation of primer efficiency in multiplex PCR for detecting SARS-Cov-2 variants. Bionatura. 2022; 7(3):48.

Al-Rubii, BAL. Cloning LasB gene of Pseudomonas aeruginosa eiastase 10104-2aI in E. coli BL21 and E.coli DH5α and investigated their effect on the stripping of vero cells. Pakistan J Biotechnol.2017; 14(4):697-705.

Saleh, T.; Hashim, S.; Malik, S.N.; Al-Rubaii, B.A.L. The impact some of nutrients on swarming phenomenon and detection the responsible gene RsbA in clinical isolates of Proteus mirabilis. Int J Pharm Sci Res. 2020;1(6):437-444.

Bresam S, Al-Jumaily RM, Karim GF, Al-Rubaii BA. Polymorphism in SNP rs972283 of the KLF14 gene and genetic disposition to peptic ulcer. Biomedicine. 2023; 43(1):216-220.

Shehab ZH, Laftah BA. Correlation of nan1 (Neuraminidase) and production of some type III secretion system in clinical isolates of Pseudomonas aeruginosa. Biomed Res. 2018;15(3):1729-1738.

Rasoul, LM., Marhoon AA, Albaayit SFA, Ali RW, Saleh,TH, Al-Rubaii BAL. Cytotoxic effect of cloned EGFP gene on NCI-H727 cell line via genetically engineered gene transfer system. Biomedicine. 2022; 42(5): 938-942.

Abdulateef Y. Role of PSA in Diagnosis of Chronic Prostatitis. Indian Journal of Forensic Medicine & Toxicology. 2020; 14(1):774-779.

Ali SM, Laftah BA, Al-Shammary AM, Salih HS. Study the role of bacterial neuraminidase against adenocarcinomacells in vivo. In AIP Conference Proceedings 2021, 2372, 030009.

Jalil IS, Mohammad SQ, Mohsen AK, Al-Rubaii BA. Inhibitory activity of Mentha spicata oils on biofilms of Proteus mirabilis isolated from burns. Biomedicine. 2023; 43(02):748-52.

Salih HS, Al-Shammari AM, Laftaah BA. Intratumoral coadministration of oncolytic newcastle disease virus and bacterial hyaluronidase enhances virus potency in tumor models. J Glob Pharma Technol. 2018; 10(10):303-310.

Abbas MS, Ahmed AG, Ali SQ, AL-Rubaii BA. Immunological inflammatory factors in patients diagnosed with COVID-19. Biomedicine. 2023; 43(1):230-23



How to Cite

Qasim, M. J., Al-Attraqchi, A. A., & Aziz, G. M. (2023). Evaluation of Cytotoxicity and Antimicrobial Activity of Flavipin Against Acinetobacter baumannii, Methicillin-Resistant Staphylococcus aureus, and Candida albicans: http://www.doi.org/10.26538/tjnpr/v7i7.2. Tropical Journal of Natural Product Research (TJNPR), 7(7), 3279–3284. Retrieved from https://tjnpr.org/index.php/home/article/view/2212