Cloning and Characterization of the UDP Glucose/Galactose Epimerases of Oryza sativa
Cloning and Characterization of the UDP Glucose/Galactose Epimerases of Oryza sativa
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DOI：
10.3839/jksabc.2009.056
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发表时间：
2009-08-01
影响因子：
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通讯作者：
Ahn, Joong-Hoon
中科院分区：
文献类型：
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作者：
Kim, Seong-Kyong;Kim, Dae Hwan;Ahn, Joong-Hoon
UDP-葡萄糖/半乳糖-4-差向异构酶(UGE)催化UDP-葡萄糖和UDP-半乳糖的相互转化。水稻的唾液中含有四种唾液。除茎外，OsUGE3在所有组织中的表达均高于其他组织。干旱、盐分或紫外线胁迫后，OsUGE1的表达均增强。利用聚合酶链式反应技术克隆了4个水稻Uu基因(OsUGE1-4)，并对其进行了序列测定。用谷胱甘肽S-转移酶融合蛋白在大肠杆菌中表达并纯化了OsUGE1-4。四种OsUGE均能更好地将UDP-葡萄糖转化为UDP-半乳糖，反之亦然。OsUGEs的动力学参数表明，OsUGE1。是最有效率的。这些OsUGE可用于各种UDP-糖的生物合成，这些UDP-糖可作为抗生素或类黄酮类UDP依赖的糖基转移酶修饰反应的辅基。
UDP-glucose/galactose-4-epimerase (UGE) catalyzes the interconversion of UDP-glucose and UDP-galactose. Oryza saliva contains four UGEs. Expression of OsUGE3 was higher than other OsUGEs in all tissues except the stem. OsUGE1 expression was enhanced after drought, salt or UV-irradiation stress. Four rice UGEs (OsUGE1-4) were cloned by polymerase chain reaction and sequenced. OsUGE1-4 were expressed in Escherichia coli with glutathione S-transferase fusion protein and purified. All four OsUGEs could covert UDP-glucose into UDP-galactose better than vice versa. Kinetic parameters of OsUGEs showed that OsUGE1. was most efficient. These OsUGEs could be used for biosynthesis of various UDP-sugars, which serve as a cosubstrates of UDP-dependent glycosyltransferase modification reactions of antibiotics or flavonoids.