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3.4. Phylogenetic Analysis of PM0525875 The 16S rRNA gene from PM0525875 was sequenced. The partial sequence containing 1377 nucleotide base pairs showed close similarity with Actinoalloteichus cyanogriseus. The phylogenic tree constructed using ten matching sequences of 16S rRNA from NCBI data-base is shown in Figure 7. The evolutionary history was inferred using the Neighbor-Joining method [20]. The bootstrap consensus tree inferred from 500 replicates was represented in the evolutionary history of the taxa [21] and the branches corresponding to partitions reproduced in less than 50% bootstrap replicates were collapsed. The evolutionary distances were computed using the Kimura 2-parameter method [22]. The analysis involved 11 nucleotide sequences and the codon positions includedwere 1st + 2nd + 3rd + Noncoding. There were a total of 1381 positions in the final datasheet. Evolutionaryanalyses were conducted on computer software MEGA5 [23]. Based on this results the isolated actinomycetes strain was identified as Actinoalloteichus cyanogriseus. The gene sequence was deposited in NCBI GenBank under accession no KF861694.1. 3.5. MIC of Caerulomycin A MIC of the isolated compound Caerulomycin A and two standard antifungal compounds was determined using NCCLS Macrobroth dilution method and results are shown in Table 6. Caerulomycin A showed potent activity against four Candida strains, including two fluconazole resistant strains. The MIC of the compound was in the range of 0.39 - 1.256 μg/ml. The MIC values obtained of Caerulomycin A against fluconazole resistant Candida glabrata were comparable with the MIC values obtained for Amphotericin B. This result indicates the potential of Caerulomycin A as a potent and broad spectrum antifungal agent. |
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