Antimicrobial and Antioxidant Activities of Secondary Metabolites of an Endophytic fungus of Azadirachta indica

Authors

  • Nkechi J. Okonkwo-Uzor Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria
  • Cynthia Obi Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria
  • Adaobi Enwelum Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria
  • Ugochukwu M. Okezie Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Science, Nnamdi Azikiwe University, Awka Anambra State Nigeria
  • Peter M. Eze Department of Environmental Health Science, Faculty of Health Sciences and Technology, Nnamdi Azikiwe University, Awka, Anambra State Nigeria
  • Festus B. C. Okoye Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi, Azikiwe University, Awka Anambra State Nigeria
  • Charles O. Esimone Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Science, Nnamdi Azikiwe University, Awka Anambra State Nigeria

Abstract

Background: Recent upsurge in the mortality rate and long hospital stays as a result of antimicrobial resistance caused by multi-drug resistant bacteria is a huge burden to public health sector. Intensive search for more effective and newer agents to deal with these problems. Endophytes are a novel source of potentially useful medicinal compounds. Aim: The study aimed at evaluating the antimicrobial and antioxidant activities of an endophytic fungus isolated from Azadirachta indica against multi drug resistant bacteria species. Method: Endophytic fungus was isolated from fresh leaves of Azadirachta indica. The fungus was fermented in rice medium, and the secondary metabolites were extracted. The antimicrobial activity of the extract against laboratory strains of Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia and Bacillus subtilis, Candida albicans, Aspergillus niger) was determined. The fungal extract was also evaluated for antioxidant activity using the DPPH assay method. Using GC-FID analysis, some constituents of the fungal extracted were identified. Result: At 1 and 0.5 mg/mL, the fungal extract demonstrated antibacterial activity against Klebsiella pneumoniae but no activity against the other test microbes. However, the fungal extract of A. indica showed excellent antioxidant activity due to possession of bioactive secondary metabolites. Conclusion: The result of GC-FID showed the presence of Rutin, Lunamarin, Anthocyanin, Catechin, Naringenin, Flavan 3 ol, Sapogernin, Tannin, Spartein, Naringin, Flavonones, Steroids, Kaemferol, Cyanogenic glycoside, Phytate, Epicatechin, Reseveratol, Cardic glycoside and Epihedrine at different concentrations. This study reveals the dynamic potentials of the endophytic fungus of A. indica as the bedrock of naturally occurring bioactive agent. 

Keywords: Azadirachta indica, endophytes, GC-FID, secondary metabolites, medicinal plants, phytochemicals.

Keywords:

Azadirachta indica, endophytes, GC-FID, secondary metabolites, medicinal plants, phytochemicals

DOI

https://doi.org/10.22270/jddt.v12i5-S.5642

Author Biographies

Nkechi J. Okonkwo-Uzor, Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria

Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria

Cynthia Obi, Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria

Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria

Adaobi Enwelum, Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria

Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences. Chukwuemeka Odumegwu Ojukwu University, Igbariam, Anambra State Nigeria

Ugochukwu M. Okezie, Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Science, Nnamdi Azikiwe University, Awka Anambra State Nigeria

Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Science, Nnamdi Azikiwe University, Awka Anambra State Nigeria

Peter M. Eze, Department of Environmental Health Science, Faculty of Health Sciences and Technology, Nnamdi Azikiwe University, Awka, Anambra State Nigeria

Department of Environmental Health Science, Faculty of Health Sciences and Technology, Nnamdi Azikiwe University, Awka, Anambra State Nigeria

Festus B. C. Okoye, Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi, Azikiwe University, Awka Anambra State Nigeria

Department of Pharmaceutical and Medicinal Chemistry, Faculty of Pharmaceutical Sciences, Nnamdi, Azikiwe University, Awka Anambra State Nigeria

Charles O. Esimone, Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Science, Nnamdi Azikiwe University, Awka Anambra State Nigeria

Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Science, Nnamdi Azikiwe University, Awka Anambra State Nigeria

References

White, J. F., Kingsley, K. L., Zhang, Q., Verma, R., Obi, N., et al.,. Review: Endophytic microbes and their potential applications in crop management. In Pest Management Science. 2019; 75(10):2558–2565). https://doi.org/10.1002/ps.5527

Al-ghamdi, A. A., Elshikh, M. S., and Hatamleh, A. A. In-vitro Phytochemical and Pharmacological bio-efficacy studies on Azadirachta indica A. Juss and Melia azedarach Linn for anticancer activity. Saudi Journal of Biological Sciences. 2019. https://doi.org/10.1016/j.sjbs.2019.11.024

Lucas, F., Batista, A., Lima, L. M. G., Abrante, I. A., Ismael, J., et al.,. Biomedicine & Pharmacotherapy Antinociceptive activity of ethanolic extract of Azadirachta indica A . Juss ( Neem , Meliaceae ) fruit through opioid, glutamatergic and acid-sensitive ion pathways in adult zebra fish (Danio rerio). Biomedicine & Pharmacotherapy. 2018; 10:408–416. https://doi.org/10.1016/j.biopha.2018.08.160

Chutulo EC, and Chalannavar RK. Endophytic mycoflora and their bioactive compounds from Azadirachta indica: a comprehensive review. Journal of Fungi. 2018; 4(2):42.

Okezie UM, Eze PM, Okoye FBC, and Esimone CO. Preliminary investigation of the chemo diversity of bioactive molecules produced by endophytic fungi isolated from Manihot utilisima leaf. GSC Biological and Pharmaceutical Sciences. 2021; 17(01):011–025. https://doi.org/10.30574/gscbps.2021.17.1.0284

Akpotu, M. O., Eze, P. M., Abba, C. C., Umeokoli, B. O., Nwachukwu, C. U et al.,. Antimicrobial activities of secondary metabolites of endophytic fungi isolated from Catharanthus roseus. Journal of Health Sciences. 2017; 7(1):15–22. https://doi.org/10.17532/jhsci.2017.421

Eze PM, Ojimba NK, Abonyi DO, Chukwunwejim CR, Abba CC et al.,. Antimicrobial Activity of Metabolites of an Endophytic Fungus Isolated from the Leaves of Citrus jambhiri (Rutaceae). Trop J Nat Prod Res. 2018; 2(3):145-149.

Okoye FBC, Nworu CS, Debbab A, Esimone CO and Proksch P. Two new Cytochalasins from an endophytic fungus, KL-1.1 isolated from Psidium guajava leaves. Phytochem Lett. 2015; 14:51-55.

Abba CC, Nduka I, Eze PM, Ujam TN, Abonyi DO et al.,. Antimicrobial Activity of Secondary Metabolites of Endophytic Aspergillus Species Isolated from Loranthus micranthus. Afr J Pharm Res Dev. 2016; 8(2):136-140.

Ebada SS, Eze P, Okoye FBC, Esimone CO, and Proksch P. The Fungal Endophyte Nigrospora oryzae Produces Quercetin Monoglycosides Previously Known Only from Plants. Chem Select. 2016; 1:2767 – 2771

Okezie UM, Eze PM, Okoye FBC, Ikegbunam MN, Ugwu MC et al.,. Biologically Active Metabolites of an Endophytic Fungus Isolated from Vernonia amygdalina. Afr J Pharm Res Dev. 2017; 9(1):24-29.

Wang H, Eze PM, Hӧfert S, Janiak C, Hartmann R, Okoye et al.,. Substituted L-tryptophan-L-phenyllactic acid conjugates produced by an endophytic fungus Aspergillus aculeatus using an OSMAC approach. RSC Adv. 2018; 8:7863–7872.

Eze PM, Nnanna JC, Okezie MU, Buzugbe SH, Abba CC, et al.,. Screening of metabolites from endophytic fungi of some Nigerian medicinal plants for antimicrobial activities. Euro Biotech Journal. 2019; 3(1). DOI: 10.2478/ebtj-2019-0002

Bezerra K S, and Antoniosi-Filho N. Characterization and Quantification by Gas Chromatography of Free Steroids in Unsaponifiable Matter of Vegetable Oils. J. Braz. Chem. Soc. 2014; 25(2):238-245.

Chigozie V, Okezie M, Ajaegbu E, Okoye FBC and Esimone CO. Isolation, identification and evaluation of biological activities of Daldinia eschscholtzii, an endophytic fungus isolated from the leaves of Musa paradisiace. GSC Biological and Pharmaceutical Sciences. 2020; 12(1):216-228.

Lugtenberg BJ, Caradus JR, and Johnson LJ. Fungal endophytes for sustainable crop production. FEMS Microbiol Ecol. 2016; 92(12):fiw194. doi: https://doi.org/10.1093/femsec/fiw194 .

Serrano J, Puupponen-Pimi R, Dauer A, Aura AM, and Sauer-Calixto F. Tannins: Current knowledge of food sources, intake, bioavailability and biological effects. Molecular Nutrition and Food Research. 2009; 53:310-329.

Ojo, O. A., Ojo, A. B., Barnabas, M., Iyobhebhe, M., Elebiyo, T. C., et al.,. Phytochemical properties and pharmacological activities of the genus Pennisetum: A review. Scientific African. 2022; 16:e01132. https://doi.org/10.1016/j.sciaf.2022.e01132

Ojukwu, S. U., Fa, O., and Ce, U. GC-FID phytochemical analysis of selected antidiabetic herbal preparations sold in Onitsha metropolis , Anambra state , Nigeria. 2021; 9(4):60–67.

Duru, C. E., and Duru, I. A. Phytochemical evaluation and health risk assessment of honey from an Apiary in Amizi, Ikuano local government area, Abia State, Nigeria. Scientific African. 2021; 13, e00885. https://doi.org/10.1016/j.sciaf.2021.e00885

Chen, R., Qi, Q. L., Wang, M. T., and Li, Q. Y. Therapeutic potential of naringin: an overview. Pharmaceutical Biology. 2016; 54(12):3203–3210. https://doi.org/10.1080/13880209.2016.1216131

Panche, A. N., Diwan, A. D., and Chandra, S. R. Flavonoids: An overview. Journal of Nutritional Science. 2016; 5. https://doi.org/10.1017/jns.2016.41

Rieko Yachi, Chie Muto, Natsuki Ohtaka, Yoshinori Aoki, Taisuke Koike, et al. Effects of tocotrienol on tumor necrosis. J. Clin. Biochem. Nutr. 2013; 52(2):146–153. https://doi.org/10.3164/jcbn.12

Mosayyebi, B., Imani, M., Mohammadi, L., Akbarzadeh, A., Zarghami, N., Edalati, M., et al.,. An update on the toxicity of cyanogenic glycosides bioactive compounds: Possible clinical application in targeted cancer therapy. Materials Chemistry and Physics. 2020; 246(February), 122841. https://doi.org/10.1016/j.matchemphys.2020.122841

Jesus, M., Martins, A. P. J., Gallardo, E., and Silvestre, S. Diosgenin : Recent Highlights on Pharmacology and Analytical Methodology. 2016.

Singh, A. K., Sharma, R. K., Sharma, V., Singh, T., Kumar, R., and Kumari, D. Isolation, morphological identification and in vitro antibacterial activity of endophytic bacteria isolated from Azadirachta indica (neem) leaves. In Veterinary World. 2017; 10(5):510–516). https://doi.org/10.14202/vetworld.2017.510-516

Linh, D. N., Thien, B. Van, Ni, T. , Duy, N. T., Minh, N. Van, et al.,. Studies on antimicrobial activities of endophytic bacteria isolated from Neem tree (Azadirachta indica). Engineering and Technology. 2020; 10(1):72–80. https://doi.org/10.46223/hcmcoujs.tech.en.10.1.362.2020

Mistry, K. S., Sanghvi, Z., Parmar, G., and Shah, S. The antimicrobial activity of Azadirachta indica , sanctum and 2 % chlorhexidine gluconate on common endodontic pathogens : An in vitro study. 2019; 8(2). https://doi.org/10.4103/1305-7456.130591

Nigussie, D., Davey, G., Legesse, B. A., Fekadu, A. and Makonnen, E. Antibacterial activity of methanol extracts of the leaves of three medicinal plants against selected bacteria isolated from wounds of lymphoedema patients. BMC Complementary Medicine and Therapies. 2021; 21(1): 2.

Daniel, I. E., Akpan, E. I., and Utam, E. C. Phytochemical Evaluation, Antioxidant and Antimicrobial Activities of Various Extracts from Leaves and Stems of Bryophyllum pinnatum. Nepal Journal of Biotechnology. 2020; 8(1):17–28. https://doi.org/10.3126/njb.v8i1.30206

Xie J, Schaich KM. Re-evaluation of the 2,2-Diphenyl-1-picrylhydrazyl free radical (DPPH) assay for antioxidant activity. J. Agric Food Chem. 2014; 62: 4251-4260.

Sinurat AP, Wina E, Rakhmani S, Wardhani T, Haryati T and Purwadaria T. Bioactive Substances of some herbals and their effectiveness as antioxidant, antibacterial and antifungi. Jurnal Ilmu Ternak dan Veteriner. 2017; 23(1):18. Doi: https://doi.org/10.14334/jitv.v23i1.1660

Published

2022-10-15
Statistics
Abstract Display: 958
PDF Downloads: 754
PDF Downloads: 32

How to Cite

1.
Okonkwo-Uzor NJ, Obi C, Enwelum A, Okezie UM, Eze PM, Okoye FBC, et al. Antimicrobial and Antioxidant Activities of Secondary Metabolites of an Endophytic fungus of Azadirachta indica. J. Drug Delivery Ther. [Internet]. 2022 Oct. 15 [cited 2026 Jan. 16];12(5-S):112-7. Available from: https://jddtonline.info/index.php/jddt/article/view/5642

How to Cite

1.
Okonkwo-Uzor NJ, Obi C, Enwelum A, Okezie UM, Eze PM, Okoye FBC, et al. Antimicrobial and Antioxidant Activities of Secondary Metabolites of an Endophytic fungus of Azadirachta indica. J. Drug Delivery Ther. [Internet]. 2022 Oct. 15 [cited 2026 Jan. 16];12(5-S):112-7. Available from: https://jddtonline.info/index.php/jddt/article/view/5642

Most read articles by the same author(s)