Preparation And Ferritin Iron Chickpea Efficacy Studies

Ferritins are iron storage proteins constructed from24subunits. Ferritins are ubiquitously expressed in animals, plants and bacteria, and possess certain properties such as soluble, non-toxic and bioavailable.They serve the dual function of iron metabolism and iron detoxification. Ferritins are the source of photosynthesis and nitrogen fixation. In contrast to animal ferritin, relatively little information is available on phytoferritin, and concentrate on legume seeds.In this paper, we used the chickpea as research object, to maximize the extraction yield of ferritin, technological parameters such as buffer type, buffer pH, solid-liquid ratio,salt type and concentration were investigated by single factor method followed by four factor-three level orthogonal experiment and response surface methodology, according to Box-Behnken principles, the response surface methodology with three factors and three levels was used. The method of orthophenanthroline staining was used to detect the ferritin. The method of braford was used to detect the protein. Under the conditions of solid-liquid ratio of1:4,50mmol/L MgCl2,KH2PO4-NaOH buffer pH7.5, the optimal extraction yield was2.659X10"5.The crude extraction of dry chickpea seed, which was obtained through homogenization, filtration and centrifugation, was precipitated by MgCl2firstly and then sodium citrate. After centrifugation and dialysis, the impure protein was chromatographed on DEAE-Sepharose Fast Flow anion-exchanger equilibrated with pH7.5,20mmol/L KH2PO4-NaOH buffer, the adsorbed substances were eluted with a linear gradient of NaCl(0.1-1.0mol/L) in the buffer. The fractions5mL were collected with a flow rate of1.0mL/min. then the sample was applied to a Sephacryl S-500gel filtration equilibrated with the buffer containing0.3mol/L NaCl. After eluted with the flow rate of0.5mL/min, the flow rate of0.1mL/min was executed. After that, the pure protein was analysed through Native-PAGE electrophoresis techniques whose molecular weight was about560kDa. Under the conditions of SDS-PAGE, it depolymerized into only one subunit of27.9kDa, which reveal that the subunit of purified ferritin of chickpea was undivided.Wistar rats received iron deficient diet, after four weeks, the rats with hemoglobin less than90g/L were selected as the iron deficiency anemia rats and divided into5groups randomly. Then the rats received deionized water, FeSO4, chickpea ferritin separately. After intervention, the rats’s blood was collected and used to do the routine blood test. As a result, the chickpea ferritin had a significant effect on treating iron deficiency anemia.