Cultural characterization was done on ISP (International Streptomyces Project) JQ1 media; yeast extract – malt extract agar (ISP-2), oatmeal agar (ISP-3), glycerol asparagine agar (ISP-5), peptone yeast extract iron agar (ISP-6), inorganic salts starch agar (ISP-4), tyrosine agar (ISP-7) and nutrient agar at 28 °C. All media were obtained from Hi-Media, Mumbai. The growth of the
organism was studied at different temperatures and salt concentrations such as 22, 28, 37, 42 °C and 2, 4, 6, 8, 10% respectively. Utilization of different carbon and nitrogen sources such as d-glucose, d-galactose, d-fructose, d-mannitol, d-xylose, l-arabinose, l-rhamnose, l-raffinose, l-cysteine, l-histidine, l-tyrosine, d-alanine, l-leucine, l-phenylalanine and l-valine was studied. Chemotaxonomic studies were done by analyzing the cells for 2,6-diaminopimelic acid.9 16S rRNA studies were conducted and isolate MS02, was submitted in Microbial Type Culture Collection, IMTECH, Chandigarh, India. The preparation of total genomic DNA was conducted in accordance with the methods described by Sambrook et al7 PCR amplification of the 16S rRNA gene of the local Streptomyces strain MS02 was conducted
in accordance with the method described by Edwards et al 10 The sequence data were deposited in the GenBank database, under the accession number JF915304. The BLAST program (www.ncbi.nlm.nih.gov/blst) was employed in order to assess the degree of DNA similarity. Multiple sequence alignment and molecular phylogeny were evaluated using
BioEdit Epigenetic inhibitor datasheet software and the phylogenetic tree was displayed using the TREE and VIEW program. 11 Spore suspension of Streptomyces isolate MS02, was prepared from the freshly grown culture on starch casein nitrate agar slant and inoculated into 100 ml starch casein nitrate broth (107 spores/ml of the medium) in 500 ml Erlenmeyer flask. The flask was incubated on rotary shaker (180 rpm) for 5 days at 28 °C. The culture was centrifuged at 8000 rpm for 20 min. The culture supernatant was used as a source of antifungal metabolite against C. albicans MTCC 183, as a target organism. Antifungal metabolite production was carried out in 100 ml starch casein nitrate broth (soluble starch – 10 g, Potassium phosphate dibasic – 2 g, Potassium nitrate – 2 g, Sodium chloride – 2 g, Casein –0.3 g, MgSO4. 7H2O – 0.05 g, CaCO3 – 0.02 g, FeSO4· 7H20 – 0.01 g, Distilled water – 1000 ml, pH – 7) in 500 ml Erlenmeyer flasks. The initial pH of the starch casein nitrate broth was adjusted to 4, 5, 6, 7, 8 and 9 separately with 0.1N NaOH/0.1N HCl. The pH 7.2 was used as control. All flasks were inoculated as mentioned above and incubated at 28 °C on rotary shaker at 180 rpm for 5 days.