Purification and characterization of lipoxygenase from Entermorpha clathrata |
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| Authors: | YANG Wenge XUE Changhu XU Dalun FU Xueyan and HE Xiong |
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| Institution: | 1.College of Food Science and Technology, Ocean University of China, Qingdao 266003, China;Faculty of Life Science and Biotechnology, Ningbo University, Ningbo 315211, China2.College of Food Science and Technology, Ocean University of China, Qingdao 266003, China3.Faculty of Life Science and Biotechnology, Ningbo University, Ningbo 315211, China4.Zhejiang Pharmaceutical College, Ningbo 315100, China |
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| Abstract: | The purification and characterization of lipoxygenase (LOX,EC 1.13.11.12) from the green algae Enteromorpha clathrata were studied.Two components marked LOX-1 and LOX-2 were purified and their molecular masses were estimated to be 102 and 79 ku by SDS-PAGE.Both LOX-1 and LOX-2 were stable over the pH wide range 6.0～10.0 and had the optimum pH of 10.0 and 8.6,at optimum temperature of 30 and 25℃,respectively.Substrate specificities of LOX-1 and LOX-2 were the greatest towards linoleic acid,followed by arachidonic acid and linolenic acid.The Michaelis constant values of LOX-1 and LOX-2 were 0.23 and 0.20 mmol/dm3 with the substrate of linoleic acid.The LOX activities were stimulated greatly by Ca2 but in- hibited by Hg2 and the antioxidants such as BHA,BHT and TBHQ.The hydroperoxide products of LOX were analysed by HPLC with the substrate of methyl linoleate,and the results showed that LOX-1 formed mainly 9-hydroperoxides while LOX-2 formed both 9-and 13-hydroperoxides at a ratio of 24:76. |
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| Keywords: | Entermorpha clathrata lipoxygenase purification characterization |
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