Zakia. A. Osman, Sulieman M. Elsanousi and Elsiddig. A. Elmustfa. Elsheikh.
In the present work, ferredoxin was extracted from the locally grown spinach plant. The spinach ferredoxin was purified through the Diethylaminoethyl (DEAE) - cellulose column chromatography at 4°C and was sterilized by filtration. Spinach ferredoxin activity was determined by the absorbency measurement (UV –Spectrophotometer) in ultraviolet (UV) - visible region, and with using the total iron content, by the Atomic Absorption Spectrometer. Different concentrations of sterilized spinach ferredoxin were added to the Reinforced Clostridial Agar (RCA) and Cooked meat Media (CMM) as substitutes for cysteine hydrochloride. The amended RCA was then used for cultivation of Clostridium perfringens strain. The growth rate of this bacterium was significantly higher when compared with that on the control medium (RCA anaerobic). The highest growth occurred with 3% spinach ferredoxin added to the RCA. Moreover, the spinach ferredoxin supported the anaerobic bacterial growth even in the lowest concentration (O.O1%). Generally, the tested anaerobic bacterium, Clostridium perfringens produced larger colonies, with regular glistening smooth surface and with irregular or entire edges on media prepared from the spinach ferredoxin, than that on the control medium. This study was targeting to evaluate the importance of spinach-ferredoxin as reducing agent in anaerobic media, liquid and solid, which is more available, cheaper and non-toxic even for human, when compared with cysteine hydrochloride.
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