National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
Unlocking cryptic secondary metabolic pathways in microorganisms remains a central challenge in natural product discovery. Under standard single-strain laboratory conditions, many biosynthetic gene clusters (BGCs) remain silent or unexpressed. This study investigates the interkingdom co-cultivation of Pseudomonas aeruginosa and Mucor sp. (PM co-culture) as a strategy to stimulate competitive stress and interspecies signaling exchanges. Ethyl acetate extracts of the PM co-culture and its respective monocultures were analyzed for chemical diversity via Gas Chromatography-Mass Spectrometry (GC-MS) and evaluated for antioxidant, anti-inflammatory, antimicrobial, anti-biofilm, and cytotoxic properties. GC-MS profiling revealed a significant metabolic shift, with the PM extract displaying unique peaks—including monoterpene alcohols (α-terpineol), phenyl ester derivatives, and siloxane compounds—that were entirely absent in single-strain controls. The co-culture extract demonstrated superior biological efficacy compared to monocultures, including notable DPPH radical scavenging (77.10%), potent inhibition of heat-induced protein denaturation (85.35%), effective disruption of microbial biofilms (41.60%), and marked cytotoxicity against MCF-7 breast cancer cells (IC50 = 4.80μg/mL). These findings confirm that bacterial-fungal co-cultivation between Pseudomonas sp. and Mucor sp. effectively activates silent metabolic pathways, presenting a powerful strategy for discovering novel bioactive compounds.
Albuquerque, P., and Casadevall, A. Quorum sensing in fungi – a review. Medical Mycology, 2012, 50(4): 337–345. https://doi.org/10.3109/13693786.2011.604851
Atlas, R. M. Handbook of Microbiological Media (4th ed.). CRC Press, 2010. https://doi.org/10.1201/EBK1439804063
Bastos, R. W., Akiyama, D., Dos Reis, T. F., Colabardini, A. C., Luperini, R. S., De Castro, P. A., et al., Secondary metabolites produced during Aspergillus fumigatus and Pseudomonas aeruginosa biofilm formation. mBio, 2022, 13(4): e01850-22. https://doi.org/10.1128/mbio.01850-22
Carducci, M., Whitcombe, A., Rovetini, L., Massai, L., Keeley, A. J., de Silva, T. I., et al., Development and characterization of a hemolysis inhibition assay to determine functionality of anti-Streptolysin O antibodies in human sera. Journal of Immunological Methods, 2024, 526: 113618. https://doi.org/10.1016/j.jim.2024.113618
Espinosa-Ortiz, E. J., Rene, E. R., and Gerlach, R. Potential use of fungal-bacterial co-cultures for the removal of organic pollutants. Critical Reviews in Biotechnology, 2022, 42(3): 361–383. https://doi.org/10.1080/07388551.2021.1940832
Eze, F. U., Okoro, U. C., Ugwu, D. I., and Okafor, S. N. Biological activity evaluation of some new benzenesulphonamide derivatives. Frontiers in Chemistry, 2019, 7: 634. https://doi.org/10.3389/fchem.2019.00634
Fleming, F. F., Yao, L., Ravikumar, P. C., Funk, L., and Shook, B. C. Nitrile-containing pharmaceuticals: efficacious roles of the nitrile pharmacophore. Journal of Medicinal Chemistry, 2010, 53(22): 7902-7917. https://doi.org/10.1021/jm100762r
Hamed, A. A., Ghareeb, M. A., Kelany, A. K., Abdelraof, M., Kabary, H. A., Soliman, N. R., et al., Induction of antimicrobial, antioxidant metabolites production by co-cultivation of two red-sea-sponge-associated Aspergillus sp. CO2 and Bacillus sp. COBZ21. BMC Biotechnology, 2024, 24(1): 3. https://doi.org/10.1186/s12896-023-00827-0
Jiang, Q., Chen, J., Yang, C., Yin, Y., and Yao, K. Quorum sensing: A prospective therapeutic target for bacterial diseases. BioMed Research International, 2019, 2019: 2015978. https://doi.org/10.1155/2019/2015978
Liu T, Gui X, Zhang G, Luo L, Zhao J. Streptomyces-Fungus Co-Culture Enhances the production of Borrelidin and Analogs: A Genomic and Metabolomic Approach. Marine Drugs 2024;22: 302. https://doi.org/10.3390/md22070302
McFarland, J. The nephelometer: An instrument for estimating the number of bacteria in suspensions used for calculating the opsonic index and for vaccines. The Journal of the American Medical Association, 1907, 49(14): 1176–1178. https://doi.org/10.1001/jama.1907.25320140022001f
McLafferty, F. W. Registry of Mass Spectral Data (10th ed.). John Wiley & Sons, 2013. https://www.semanticscholar.org/paper/The-Wiley-NBS-registry-of-mass-spectral-data-McLafferty-Stauffer/8047db76d3244204520d2e809a5a7787e62e9c24
Mizushima, Y., and Kobayashi, M. Interaction of anti-inflammatory drugs with serum proteins, especially with some biologically active proteins. Journal of Pharmacy and Pharmacology, 1968, 20(3): 169–173. https://doi.org/10.1111/j.2042-7158.1968.tb09718.x
Mohamed, H., Awad, M. F., Shah, A. M., Sadaqat, B., Nazir, Y., Naz, T., et al., Coculturing of Mucor plumbeus and Bacillus subtilis bacterium as an efficient fermentation strategy to enhance fungal lipid and gamma-linolenic acid (GLA) production. Scientific Reports, 2022, 12(1): 13111. https://doi.org/10.1038/s41598-022-17181-4
Molyneux, P. The use of the stable free radical diphenylpicrylhydrazyl (DPPH) for estimating antioxidant activity. Songklanakarin Journal of Science and Technology, 2003, 26(2): 211–219. https://www.researchgate.net/publication/237620105_The_use_of_the_stable_radical_Diphenylpicrylhydrazyl_DPPH_for_estimating_antioxidant_activity
Mosmann, T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 1983, 65(1–2): 55–63. https://doi.org/10.1016/0022-1759(83)90303-4
Moussa, M., Ebrahim, W., Kalscheuer, R., Liu, Z., and Proksch, P. Co-culture of the bacterium Pseudomonas aeruginosa with the fungus Fusarium tricinctum induces bacterial antifungal and quorum sensing signaling molecules. Phytochemistry Letters, 2020, 36: 37–41. https://doi.org/10.1016/j.phytol.2020.01.014
NIST Office of Data and Computing. Cyclononasiloxane, octadecamethyl-. NIST Chemistry WebBook, SRD 69, 2025. https://doi.org/10.18434/T4D303
Peeters, E., Nelis, H. J., and Coenye, T. Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates. Journal of Microbiological Methods, 2008, 72(2): 157–165. https://doi.org/10.1016/j.mimet.2007.11.010
PubChem. Phosphonic acid, ethyl-, bis(trimethylsilyl) ester (CID 536413). National Center for Biotechnology Information, 2025. https://pubchem.ncbi.nlm.nih.gov/compound/536413
Reynolds, J. Serial Dilution Protocols. American Society for Microbiology (ASM) MicrobeLibrary, 2005. https://asm.org/protocols/serial-dilution-protocols
Rodriguez, C., Pauli, W., and Bazevque, P. An antibiotic assay by the agar well diffusion method. Journal of Microbiological Methods, 1990, 12(3-4): 205–211. https://doi.org/10.1016/0167-7012(90)90032-N
Sanitá Lima, M., and Coutinho De Lucas, R. Co-cultivation, co-culture, mixed culture, and microbial consortium of fungi: An understudied strategy for biomass conversion. Frontiers in Microbiology, 2022, 12: 837685. https://doi.org/10.3389/fmicb.2021.837685
Schroeckh, V., Scherlach, K., Nützmann, H. W., Shelest, E., Schmidt-Heck, W., Schuemann, J., et al., Intimate bacterial–fungal interaction triggers biosynthesis of archetypal polyketides in Aspergillus nidulans. Proceedings of the National Academy of Sciences USA, 2009, 106(34): 14558–14563. https://doi.org/10.1073/pnas.0901870106
Shin D, Byun WS, Moon K, Kwon Y, Bae M, Um S, et al., Coculture of Marine Streptomyces sp. With Bacillus sp. Produces a New Piperazic Acid-Bearing Cyclic Peptide. Front Chem 2018;6: 498. https://doi.org/10.3389/fchem.2018.00498
Soule, H. D., Vazquez, J., Long, A., Albert, S., and Brennan, M. A human cell line from a pleural effusion derived from a breast carcinoma. Journal of the National Cancer Institute, 1973, 51(5): 1409–1416. https://doi.org/10.1093/jnci/51.5.1409
Sung A, Gromek S, Balunas M. Upregulation and Identification of Antibiotic Activity of a Marine-Derived Streptomyces sp. Via Co-Cultures with Human Pathogens. Marine Drugs 2017;15: 250. https://doi.org/10.3390/md15080250
Vashistha, A., Sharma, N., Nanaji, Y., Kumar, D., Singh, G., Barnwal, R. P., et al., Quorum sensing inhibitors as therapeutics: Bacterial biofilm inhibition. Bioorganic Chemistry, 2023, 136: 106551. https://doi.org/10.1016/j.bioorg.2023.106551
Vishali, S., and Soundhari, C. Inhibition of quorum sensing mediated pigment production in Serratia marcescens by secondary metabolite of Aspergillus sp. identified by metagenomic studies. The Pharma Innovation Journal, 2019, 8(5): 290–295. https://www.thepharmajournal.com/archives/2019/vol8issue6/PartF/8-5-155-838.pdf
Wakefield, J., Hassan, H. M., Jaspars, M., Ebel, R., and Rateb, M. E. Dual induction of new microbial secondary metabolites by fungal bacterial co-cultivation. Frontiers in Microbiology, 2017, 8: 1284. https://doi.org/10.3389/fmicb.2017.01284
Wise, K. Preparing Spread Plates Protocols. American Society for Microbiology (ASM) MicrobeLibrary, 2006. https://www.semanticscholar.org/paper/Preparing-Spread-Plates-Protocols-Wise/9cec72000efc7a89b0f88d457fac4775fdaf5333
Xu, S., Li, M., Hu, Z., Shao, Y., Ying, J., and Zhang, H. The potential use of fungal co-culture strategy for discovery of new secondary metabolites. Microorganisms, 2023, 11(2): 464. https://doi.org/10.3390/microorganisms11020464
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