Volume 14, Issue 2 (2-2026)                   JoMMID 2026, 14(2): 87-97 | Back to browse issues page

Ethics code: 31/IAEC-II/SLSPL/2024

XML Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:
Peethala M, Ajmera S, Ajmeera D, Burgula S. Bioactive Potential and HR-LC-MS Analysis of Malbranchea cinnamomea Extract. JoMMID 2026; 14 (2) :87-97
URL: http://jommid.pasteur.ac.ir/article-1-881-en.html
Bioactive Potential and HR-LC-MS Analysis of Malbranchea cinnamomea Extract
Madhuri Peethala , Shanthipriya Ajmera * , Divya Ajmeera , Sandeepta Burgula
Department of Microbiology, Palamuru University, Mahabubnagar 509001, Telangana, India
Abstract:   (5 Views)
Introduction: Thermophilic coprophilous fungi inhabiting nutrient-rich environments with intense microbial competition, such as Malbranchea cinnamomea, remain underexplored as sources of bioactive secondary metabolites. Therefore, this study aimed to evaluate the bioactive potential of M. cinnamomea ethyl acetate crude extract. Methods: M. cinnamomea was isolated from herbivore dung (goat, cattle, and sheep) and cultivated via submerged fermentation. The ethyl acetate crude extract was assessed for antibacterial activity against Staphylococcus aureus MTCC 96 via the agar well diffusion assay, cytotoxicity in RAW 264.7 murine macrophage cells using the MTT assay, and hemolytic potential through an erythrocyte lysis assay. Additional screening evaluated activity against Mycobacterium tuberculosis, Plasmodium falciparum, and dengue virus serotype 2 (DENV-2), along with cytotoxicity testing in Vero cells. Preliminary chemical characterization was performed using thin-layer chromatography (TLC), Fourier-transform infrared (FTIR) spectroscopy, and high-resolution liquid chromatography-mass spectrometry (HR-LC-MS). Results: The extract exhibited concentration-dependent cytotoxicity against RAW 264.7 cells (IC50 = 14.9 ± 0.1 μg/mL) and antibacterial activity against S. aureus, with inhibition zones of 37.7 ± 1.1 mm at 90 μg/mL. Hemolysis remained below 20% across tested concentrations, indicating low hemolytic potential. Screening showed negligible activity against M. tuberculosis, P. falciparum, and DENV-2, and no cytotoxicity was observed in Vero cells (CC50 > 1000 μg/mL). TLC revealed multiple components, FTIR revealed diverse functional groups, and HR-LC-MS detected six putative fungal metabolites including lipid derivatives, peptides, and phenolic compounds. Conclusion: M. cinnamomea produces chemically diverse secondary metabolites exhibiting antibacterial activity, concentration-dependent cytotoxicity against RAW 264.7 macrophages, and low hemolytic potential. These findings support further bioassay-guided fractionation and chemical characterization to identify novel bioactive compounds.
Keywords: Antimicrobial resistance, Bioactive metabolites, Drug discovery, HR-LC-MS, Malbranchea cinnamomea, Thermophilic fungi
Full-Text [PDF 1327 kb]   (1 Downloads)    
Type of Study: Original article | Subject: Anti-microbial agents, resistance and treatment protocols
Received: 2026/09/7 | Accepted: 2026/02/21 | Published: 2026/09/7
attachement [JPG 130 KB]  (0 Download)
attachement [PDF 2409 KB]  (0 Download)
attachement [PDF 106 KB]  (0 Download)
References
1. World Health Organization. Global antimicrobial resistance and use surveillance system (GLASS) report [Internet]. Geneva: World Health Organization; 2022 [cited 2026 Jul 31]. Available from: https://www.who.int/glass
2. Murray CJL, Ikuta KS, Sharara F, Swetschinski LR, Robles Aguilar G, Lang G, et al. Global burden of bacterial antimicrobial resistance in 2019: a systematic analysis. Lancet. 2022; 399 (10325): 629-55. [DOI:10.1016/S0140-6736(21)02724-0] [PMID]
3. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide. CA Cancer J Clin. 2021; 71 (3): 209-49. [DOI:10.3322/caac.21660] [PMID]
4. Newman DJ, Cragg GM. Natural products as sources of new drugs over nearly four decades from 1981 to 2019. J Nat Prod. 2020; 83 (3): 770-803. [DOI:10.1021/acs.jnatprod.9b01285] [PMID] [PMCID]
5. Atanasov AG, Zotchev SB, Dirsch VM, Supuran CT, International Natural Product Sciences Taskforce. Natural products in drug discovery: advances and opportunities. Nat Rev Drug Discov. 2021; 20 (3): 200-16. [DOI:10.1038/s41573-020-00114-z] [PMID]
6. Bills GF, Gloer JB. Biologically active secondary metabolites from fungi. Nat Prod Rep. 2016; 33 (7): 790-809.
7. Wadhwa K, Kapoor N, Kaur H, Abu-Seer EA, Tariq M, Siddiqui S, et al. A comprehensive review of the diversity of fungal secondary metabolites and their emerging applications in healthcare and environment. Mycobiology. 2024; 52 (6): 335-87. [DOI:10.1080/12298093.2024.2416736] [PMID] [PMCID]
8. Turner P, Mamo G, Karlsson EN. Potential and utilization of thermophiles and thermostable enzymes in biorefining. Microb Cell Fact. 2007; 6: 9. [DOI:10.1186/1475-2859-6-9] [PMID] [PMCID]
9. Cooney DG, Emerson R. Thermophilic fungi: an account of their biology, activities and classification. San Francisco: WH Freeman; 1964.
10. Pitt JI, Hocking AD. Fungi and food spoilage. 3rd ed. New York: Springer; 2009. [DOI:10.1007/978-0-387-92207-2_2] [PMID]
11. Doyle JJ, Doyle JL. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull. 1990; 19: 11-5.
12. White TJ, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, editors. PCR protocols: a guide to methods and applications. San Diego: Academic Press; 1990. p. 315-22. [DOI:10.1016/B978-0-12-372180-8.50042-1] [PMID] [PMCID]
13. Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74 (12): 5463-7. [DOI:10.1073/pnas.74.12.5463] [PMID]
14. Demain AL, Sanchez S. Microbial drug discovery: 80 years of progress. J Antibiot (Tokyo). 2009; 62 (1): 5-16. [DOI:10.1038/ja.2008.16] [PMID] [PMCID]
15. Keller NP, Turner G, Bennett JW. Fungal secondary metabolism-from biochemistry to genomics. Nat Rev Microbiol. 2005; 3 (12): 937-47. [DOI:10.1038/nrmicro1286] [PMID]
16. BerdY J. Bioactive microbial metabolites. J Antibiot (Tokyo). 2005; 58 (1): 1-26. [DOI:10.1038/ja.2005.1] [PMID]
17. Nielsen JC, Nielsen J. Development of fungal cell factories for the production of secondary metabolites. Biotechnol Lett. 2017; 39 (1): 1-14. [DOI:10.1016/j.synbio.2017.02.002] [PMID] [PMCID]
18. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65 (1-2): 55-63. [DOI:10.1016/0022-1759(83)90303-4] [PMID]
19. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. 33rd ed. CLSI supplement M100. Wayne, PA: CLSI; 2023.
20. Evans BC, Nelson CE, Yu SS, Beavers KR, Kim AJ, Li H, et al. Ex vivo red blood cell hemolysis assay for the evaluation of nanoparticle cytotoxicity. Nat Protoc. 2013; 8 (9): 1708-15.
21. Malagoli D. A full-length protocol to test hemolytic activity of compounds on human erythrocytes. MethodsX. 2016; 3: 92-7.
22. Wagner H, Bladt S. Plant drug analysis: a thin layer chromatography atlas. 2nd ed. Berlin: Springer; 1996. [DOI:10.1007/978-3-642-00574-9]
23. Coates J. Interpretation of infrared spectra, a practical approach. In: Meyers RA, editor. Encyclopedia of analytical chemistry. Chichester: John Wiley & Sons; 2000. p. 10815-37. [DOI:10.1002/9780470027318.a5606]
24. Silverstein RM, Webster FX, Kiemle DJ, Bryce DL. Spectrometric identification of organic compounds. 8th ed. Hoboken: John Wiley & Sons; 2014.
25. Smith CA, O'Maille G, Want EJ, Qin C, Trauger SA, Brandon TR, et al. METLIN: a metabolite mass spectral database. Ther Drug Monit. 2005; 27 (6): 747-51. [DOI:10.1097/01.ftd.0000179845.53213.39] [PMID] [PMCID]
26. Wang M, Carver JJ, Phelan VV, Sanchez LM, Garg N, Peng Y, et al. Sharing and community curation of mass spectrometry data with Global Natural Products Social Molecular Networking. Nat Biotechnol. 2016; 34 (8): 828-37. [DOI:10.1038/nbt.3597] [PMID]
27. Smilkstein M, Sriwilaijaroen N, Kelly JX, Wilairat P, Riscoe M. Simple and inexpensive fluorescence-based technique for high-throughput antimalarial drug screening. Antimicrob Agents Chemother. 2004; 48 (5): 1803-6. [DOI:10.1128/AAC.48.5.1803-1806.2004] [PMID] [PMCID]
28. Diamond MS, Pierson TC. Molecular insight into dengue virus pathogenesis and its implications for disease control. Cell. 2015; 162 (3): 488-92. [DOI:10.1016/j.cell.2015.07.005] [PMID] [PMCID]
29. Aron AT, Gentry EC, McPhail KL, Nothias LF, Busby B, Clark TN, et al. Reproducible molecular networking of untargeted mass spectrometry data using GNPS. Nat Protoc. 2020; 15 (4): 1954-91. [DOI:10.1038/s41596-020-0317-5] [PMID]
30. Wolfender JL, Marti G, Thomas A, Bertrand S. Current approaches and challenges for the metabolite profiling of complex natural extracts. J Chromatogr A. 2015; 1382: 136-64. [DOI:10.1016/j.chroma.2014.10.091] [PMID]
31. Li JW, Vederas JC. Drug discovery and natural products: end of an era or an endless frontier? Science. 2009; 325 (5937): 161-5. [DOI:10.1126/science.1168243] [PMID]
32. Ayon NJ, Earp CE, Gupta R, Butun FA, Clements AE, Lee AG, et al. Bioactivity-driven fungal metabologenomics identifies antiproliferative stemphone analogs and their biosynthetic gene cluster. Metabolomics. 2024; 20(5):90 [DOI:10.1007/s11306-024-02153-8] [PMID] [PMCID]
33. Demain AL. Importance of microbial natural products and the need to revitalize their discovery. J Ind Microbiol Biotechnol. 2014; 41 (2): 185-201. [DOI:10.1007/s10295-013-1325-z] [PMID]
34. Deshmukh SK, Prakash V, Ranjan A. Marine fungi: a source of potential antibacterial compounds. Front Microbiol. 2018; 9: 170. [DOI:10.3389/fmicb.2017.02536] [PMID] [PMCID]
35. Hyde KD, Xu J, Rapior S, Jeewon R, Lumyong S, Ngee-Ahing T, et al. The amazing potential of fungi: 50 ways we can exploit fungi industrially. Fungal Divers. 2019; 97 (1): 1-136. [DOI:10.1007/s13225-019-00430-9]
36. Keller NP. Fungal secondary metabolism: regulation, function and drug discovery. Nat Rev Microbiol. 2019; 17 (3): 167-80. [DOI:10.1038/s41579-018-0121-1] [PMID] [PMCID]
37. Gonçalves MF, Esteves AC, Alves A. Fungal secondary metabolites: ecological functions and applications. Appl Microbiol Biotechnol. 2023; 107 (2-3): 995-1012.
38. Prakash S, Kumari H, Sinha M, Kumar A. Regulation and induction of fungal secondary metabolites: a comprehensive review. Arch Microbiol. 2025;207(8):189 [DOI:10.1007/s00203-025-04386-0] [PMID]
39. Harvey AL, Edrada-Ebel R, Quinn RJ. The re-emergence of natural products for drug discovery in the genomics era. Nat Rev Drug Discov. 2015; 14 (2): 111-9. [DOI:10.1038/nrd4510] [PMID]
Add your comments about this article : Your username or Email:
CAPTCHA
Send email to the article author

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.