Ferric Ammonium Citrate with High Quality

• CAS No.: 1185-57-5
• Color: Brown
• Appearance: Powder
• Transport Package: Paper
• Specification: large
• Trademark: china
• Model NO.: E1422
• Origin: China
• HS Code: 2930400000
• Production Capacity: 5000kg/Year

Product Description

A B. subtilis 168 wild-type strain 1A2 (BGSC) is grown in SMS minimal medium (Spizizen, 1958) with 0.75% fructose, supplemented with 0.1% l-glutamine, 0.2 mM l-tryptophan, 22 mg/mL ferric ammonium citrate (CAF). Three liters of culture are inoculated to an OD₆₀₀ = 0.05 and grown overnight at 37°C (final OD₆₀₀ = ~ 1.2). Cells were pelleted by centrifugation (5200 × g, 15 min, 4°C), washed three times in 1% NaCl, and stored frozen at − 20°C until further use. Cell pellets were lyophilized overnight and stored at − 80°C. The total output is ~ 1 g of dried cells. 200 mg of the lyophilized cells were crushed to a coarse powder with a spatula and transferred to a separation funnel. The following solvents were added in the given order with vigorous shaking in a separating funnel before addition of the next solvent: 60 mL  120 mLand 48 mL H₂O. The funnel was kept under a hood at room temperature for 2 days with occasional shaking. 60 mL and 30 mL H₂O were added, the mixture was vigorously shaken and left for 1 day under the hood to allow for phase separation. The lower phase was recovered and the upper phase extracted with 30 mL  The lipids from the combined phases were recovered by rotary evaporation under reduced pressure (Buchi) for 10 min at 30°C. The lipid precipitate was dissolved in 5 mL  and recovered by another round of evaporation. Total lipid yield is ~ 40 mg.