摘要
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 O. 20 mmol/dm^3 with the substrate of linoleic acid. The LOX activities were stimulated greatly by Ca^2＋ but inhibited 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.
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 O. 20 mmol/dm^3 with the substrate of linoleic acid. The LOX activities were stimulated greatly by Ca^2＋ but inhibited 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.
基金
The Technology Research and Development Program ("863"Program) of China under contract No.2003AA625030.
