1252 UV SPECTROPHOTOMETRY lipoxygenase activity in wheat germ.docVIP

 PAGE \* MERGEFORMAT 8 1252 UV SPECTROPHOTOMETRY lipoxygenase activity in wheat germ Of: Xu Bin, Liu Yun Dong Jun Ying Hu Qingsong Yu Yabin [Keywords:] UV spectrophotometry lipoxygenase activity in wheat germ 1 Introduction Wheat germ (WG) in the lipoxygenase (LOX) catalyzes the oxidation of linoleic acid in the WG, the WG short term rancidity deterioration. To extend the shelf life, often in the passive heating LOX. In order to maximize the protection of WG nutrition, heating temperature and time of choice is particularly important. The accurate determination of LOX activity is the stabilization of wheat germ theory research. research shows that the transparency of the substrate solution, the reaction buffer pH, reaction temperature, substrate, and Tween concentration and other factors will affect the determination of the results. Determination of the existing cumbersome process of enzyme purification. In addition, different sources of LOX optimum pH and optimum temperature has a big difference, a choice should be based on the type of raw materials. To this end, the establishment of WG in a LOX improved detection methods, simple and fast access to a transparent substrate solution, eliminating the need for complicated purification of crude enzyme steps. 2 Experimental part 2.1 Instruments and reagents DU800 nucleic acid protein analyzer (Beckman Corporation); BR4 refrigerated centrifuge (Jouan Corporation); pHS 3B precision pH meter (Shanghai Lei magnetic Instrument Factory). Wheat germ (South flour company); linoleic acid standard ( Fluka Company); other reagents were of analytical grade. 2.2 Sample preparation to take 10 g wheat germ, grinding, adding 100 mL 0.1 mol / L acetate buffer (pH 4.5), 4 ℃ under stirring 30 min, suspension, 12000 r / min centrifugation 30 min, to obtain enzyme extract. With acetate buffer solution diluted 10 times, measured as enzyme solution. 2.3 Preparation of the substrate to take 111 μL of linoleic acid, e