Extraction, Purification and Characterization of Lipoxygenase from Aspergillus niger
- سال انتشار: 1397
- محل انتشار: کنگره توسعه همکاری های علمی منطقه ای علوم صنایع غذایی و کشاورزی
- کد COI اختصاصی: CFAS01_099
- زبان مقاله: انگلیسی
- تعداد مشاهده: 322
نویسندگان
Dept. of Food Science, College of Agriculture, University of Basra, Basrah, Iraq
Dept. of Food Science, College of Agriculture, University of Basra, Basrah, Iraq
Dept. of Food Science, College of Agriculture, University of Basra, Basrah, Iraq
چکیده
Lipoxygenase was extracted from a local isolate of Aspergillus niger that was obtained from maize. The isolate was grown and biomass was harvested after 5days of incubation at 30°C for detecting enzymatic activity. Three steps were used to purify lipoxygenase: concentration by using ammonium sulfate with (30-90%) saturation , then ion exchange chromatography by using DEAE Sephadex A-50 with NaCl (0-1)N. Two peaks appeared at the washing step and four peaks appeared at the elution step, Only one peak had specific activity ,which was 592.14 u/mg, That peak was chosen for the third step of purification, using gel filtration chromatography with Sephadex G-100. Only one peak showed specific activity (1069.7u/mg). The purity of enzyme was tested by gel electrophoresis using polyacrylamide, The enzyme was show to be purified by appearance as band. The characteristics of purified lipoxygenase were then studied: The molecular weight was104 KDa as shown by SDS- polyacrylamide gel electrophoresis, The optimum temperature and pH of enzyme activity were 35 ̊Cand 6.5 respectively. Heat stability and optimum pH of stability were (0-45) ̊C and (7-6) respectively. The effect of some mineral ions on enzymatic activity, had been noticed that CaCl2, KCl, NiCl2 , MgCl2 and MnCl2 increased the activity while the enzymatic activity was decreased when using ZnCl2,FeCl3 , and CuCl2. EDTA inhibited the enzyme activity. Studying of kinetic constants showed that Michaelis constant (Km) and maximum velocity (Vmax) of the enzyme were 0.2 mg/mLand 45 µm/mL/min respectively. The activation energy was 8 kcal/mole, and the denaturation energy was 54 kcal/mole.کلیدواژه ها
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