ABSTRACT: 44321-A2 was streaked onto R2YE agar containing 5 g of yeast extract, 103 g of sucrose, 10 g of dextrose, 0.1 g of casamino acid, 0.25 g of potassium sulfate (K2SO4), 10.12 g of magnesium chloride hexahydrate (MgCl2.6H2O), 5.73 g of TES buffer, 2 mL of trace element solution (containing 10 mg of ammonium molybdate tetrahydrate ((NH4)6Mo7O24.4H2O), 10 mg of sodium tetraborate decahydrate (Na2B4O7.10H2O), 10 mg of manganese chloride tetrahydrate (MnCl2.4H2O), 10 mg of cupric chloride dihydrate (CuCl2.2H2O), 200 mg of ferric chloride hexahydrate (FeCl3.6H2O), 40 mg of zinc chloride (ZnCl2), and 1 L of deionized water, filter sterilized), 10 mL of 0.5% potassium dihydrogen phosphate (KH2PO4), 4 mL of 5 M calcium chloride dihydrate (CaCl2.2H2O), 5 mL of 20% L-proline, 7 mL of 1 N sodium hydroxide (NaOH), 25 micrograms/mL nalidixic acid, 10 micrograms/mL benomyl, 15 g of agar, and 1 L of double distilled water. Plates were incubated for 5-7 days at 28 C. For the strain, 3 mL seed cultures in 14 mL dual position cap tubes were inoculated with a loopful of vegetative cells from R2YE plates and incubated for 5 days at 28 C, 200 rpm; 3 mL seed cultures were then inoculated into 100 mL seed cultures in 250 mL baffled flasks and incubated for 7 days at 28 C, 200 rpm; 50 mL of seed cultures were inoculated into 1 liter of fermentation media in 2.8 liter baffled Fernbach flasks and grown for 7 days at 28 C, 200 rpm.
Ashu Tripathi 
