The pH was adjusted to pH seven. 2 prior to sterilization. KM five medium consisted of four g yeast ex tract, ten g malt extract, four g glucose, 20 g agar per liter un distilled water. The pH was adjusted to pH 5. five prior to sterilization. DSMZ1 medium consisted of five g Bacto peptone, 3 g malt extract, ten mg MnSO4 x H2O and twenty g agar per liter of un distilled water. The pH was adjusted to 5. five just before sterilization. MM1 medium consisted of five g glucose, 0,5 g tri sodium citrate x 2 H2O, three g KH2PO4, 7 g K2HPO4, 0. 1 g MgSO4 x seven H2O, 1 g 2SO4 and 15 g Bacto agar. The bacteria have been cultivated for a period of 24 h in one hundred ml in respective liquid media in 500 ml Erlenmeyer flasks with 1 baffle at 27 C or 37 C on the rotary shaker at120 rpm. The cultures were centrifuged, re suspended in saline, and set to realize an optical density of 1. three at a wavelength of 546 nm.
Within the case order b-AP15 of minimum medium, cultures have been washed a single time with saline to obtain rid of complex media utilized for inoculation. Two hundred ml of complex medium containing agar were inoculated with two ml of this defined suspension of organisms. 10 ml of inoculated agar have been poured into each and every Petri dish. Strep tomyces pure culture filtrate or organic extract was find more information utilized on paper discs and air dried. The paper discs were then placed around the previ ously prepared agar media. Soon after 24 h, microbial growth inhibition was recorded by measuring the diameter with the inhibition zone. Fermentation of streptomycetes for your analysis of secondary metabolites The strains AcM9, AcM11, AcM20, AcM29 and AcM30 were cultivated in one hundred ml ISP 2 medium at 120 rpm and 27 C for 3 days. Of these cultures, 4 ml were utilized to inoculate 100 ml SGG, OM and MMN medium in 500 ml Erlenmeyer flasks with a single baffle. SGG medium consisted of 10 g soluble starch, ten g glucose, ten g gly cerol, 2.
five g cornsteep powder, 5 g Bacto peptone, two g yeast extract, 1 g NaCl and three g CaCO3 per liter of tap water. The pH was adjusted to pH 7. 3 prior to sterilization. OM medium consisted of 20 g oat meal and 5 ml of your following micronutrient solu tion, 3 g CaCl2x2 H2O, one g iron III citrat, 200 mg MnSO4 x one H2O, a hundred mg ZnCl2, 25 mg CuSO4 x 5H2O, twenty mg Na2B4O7 x 10 H2O, 4 mg CoCl2 x 6H2O, and ten mg Na2MoO4 x two H2O per liter of deionized water. The pH was adjusted to pH seven. 3 prior to sterilization. Modified MMN medium was ready in accordance to Molina and Palmer. Fermentations were carried out on the rotary shaker at 120 rpm and 27 C. After two, four and 6 days ten ml of bacterial culture had been centrifuged and bacterial biomass was determined. The culture filtrate separated from your bacterial mycelium by centri fugation was utilized for further analyses of secreted bac terial metabolites. Extraction and HPLC UV visible spectral examination of Streptomyces secondary metabolites Culture filtrates of AcM 9, AcM11, AcM20, AcM29 and AcM30 were adjusted to pH 5 and extracted with 5 ml ethyl acetate for 30 min beneath shaking condi tions.