ANTIFUNGAL ACTIVITY OF FIVE STRAINS OF SOIL MICROORGANISMS ISOLATED FROM TANGKUBAN PERAHU MOUNTAIN
Objective: The objective of this research was production of antifungal by microorganism isolated from soil of Tangkuban Perahu mountain.
Methods: The microorganisms were isolated from soil by the serial dilution method and identified by observing morphological, biochemical and molecular characteristics.
Production of antifungal metabolites was done by fermentation. Screening for bioactivity of isolates was done by testing the fermentation broth against Microsforum gypseum, the metabolite was extracted with ethyl acetate. Antifungal activities from the ethyl acetate extract were tested by agar diffusion and microdillution methods.
Results: Five strains of microorganisms were isolated from soil taken from Tangkuban Perahu mountain and were investigated in this study, designated TP1, TP2, TP3, TP4, and TP5, respectively. These isolates were shown to be closely related to Nocardia sp. YIM 65630 GU367167.1 (90%), Streptomyces galbus NR 026178.1 (99%), Aspergillus unguis FJ878626 (86%), Paecilomyces marquandii FJ765026 (100%), and Nocardia niigatensis NR 043916.1 (95%). The result of the bioactive screening showed that all isolates had antifungal activities, with the range of diameter of inhibition of 7-18 mm against M. gypseum, and TP2 has diameter of inhibition of 18 Â± 0.05 mm.
Conclusion: The highest antifungal activity showed by TP2 strain. The ethyl acetate extract of TP2 strain against Microsporum gypseum has the MIC of 4.7 Âµg/ml and diameter of inhibition was 36 Â± 0.10 mm.
2. Boca Raton FL. Anticancer Agents from Natural Products. 2nd ed. New York, CRC Press; 2005. p. 23-46.
3. Wardani IGAAK, Andayani DGS, Sukandar U, Sukandar EY. Study on microbial activity of Nocardia sp. strain TP1 isolated from Tangkuban Perahu soil, West Java, Indonesia. Int J Pharm Pharm Sci 2013;5(2):713-6.
4. Andayani DGS, Sukandar EY, Sukandar U, Adnyana IK. Isolation, identification of phenylacetic acid from Streptomyces galbus TP2 strain and its toxicity. Int J Pharm Pharm Sci 2014;6(5):643-6.
5. Demain AL, Solomon NA. Manual of Industrial Microbiology and Biotechnology. Washington DC, ASM Press; 1986.
6. Holt JG, Krieg NR, Sneath PHA, Staley JT, William ST. Bergeyâ€™s Manual of Determinative Bacteriology. Maryland, Williams and Wilkins, 9thed; 1994.
7. Cappuccino JG, Sherman N. Microbiology, A Laboratory Manual. San Francisco, Pearson Benjamin Cummings, 8th ed; 2005.
8. Pitcher DG, Saunders NA, Owen RJ. Rapid extraction of bacterial genomic DNA with guanidiumthiocyanate. Lett Appl Microbiol 1989;8:109-14.
9. Hiraishi A, Kamagata Y, Nakamura N. Polymerase chain reaction amplification and restriction fragment length polymorphism analysis of 16S rRNA genes from methanogens. Ferment Bioeng 1995;79:523-9.
10. Tamura T, Hayakawa M, Hatano K. A new genus of the order actinomycetales, spidliplanesgen. nov with description of Spirilliiplanesyamanashiensis sp nov. Int J Syst Bacteriol 1997;97-102.
11. Thompson JD, Gibson TJ, Plewniak F, Higgins DG. Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. J Nucleic Acids Res 1997;25:4876-82.
12. Felsenstein J. Confidence limitson phylogenies: an approach using the bootstrap. J Organism Evolution 1985;39:783-9.
13. Saitou N, Nei M. The neighbor-joining method: a new method for reconstruction phylogenetic tree. J Mol Biol Evol 1987;4:406-25.
14. Sukandar EY, Sastramihardja I, Padmawinata K, Satiadarma K. Microbial utilization of renewable resources. JSPS-NRCT Seminar Biotechnol 1984;4:244-51.
15. Michael LS, Fikret K. Bioprocess Engineering. New York, Prentice Hall International Series in the Physical and Chemical Engineering Sciences. 2nd ed; 2002.
16. Satyajit DS, Zahid L, Alexander IG. Natural Products Isolation. New Jersey, Humana Press, 2nd ed; 2006.
17. Mosmann T. Rapid colorimetric assay for cellular growth & survival, Application to proliferation & cytotoxicity assays. J Immunol Method 1983;65:65-59.
18. Freimoser FM, Jakob CA, Aebi M, Tuor U. The MTT ï€ ïƒ3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromideïƒ½ assay is a fast and reliable method for colorimetric determination of fungal cell densities. Appl Environ Microbiol 1999;65:3727-9.
19. Omura S. The search for bioactive compounds from microorganism. Berlin, Springer; 1992.
20. Paul AK, Banerjee AK. A new antifungal antibiotic produced by Streptomyces galbus. Folia Microbiol 1983;28:386-96.
21. Streptomyces galbus strain with insecticidal activity and method of using as an insecticide. Europesn Patent EP1272611.