Metabolic engineering of Escherichia coli to produce (2S, 3R, 4S)-4-hydroxyisoleucine
Metabolic engineering of Escherichia coli to produce (2S, 3R, 4S)-4-hydroxyisoleucine
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DOI：
10.1007/s00253-010-2772-3
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发表时间：
2010-10-01
影响因子：
5
通讯作者：
Shimizu, Sakayu
中科院分区：
文献类型：
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作者：
Smirnov, Sergey V.;Kodera, Tomohiro;Shimizu, Sakayu
在缺乏α-酮戊二酸脱氢酶（EC 1.2.4.2）、异柠檬酸连接酶（EC www.example.com）和异柠檬酸脱氢酶激酶/磷酸酶（EC 4.1.3.1）的活性的大肠杆菌2Delta菌株中克隆和表达立体特异性的1-异亮氨酸-4-羟化酶（1-异亮氨酸双加氧酶（IDO））。2.7.11.5 2 Delta菌株不能在最低盐/葡萄糖/甘油培养基中生长，这是由于在琥珀酸合成期间TCA的阻断。2 Delta菌株中的IDO活性能够“分流”被破坏的TCA，从而偶联l-异亮氨酸羟基化和细胞生长。使用该菌株，我们将l-异亮氨酸直接生物转化为4-HIL，产率为82%。
The stereo-specific l-isoleucine-4-hydroxylase (l-isoleucine dioxygenase (IDO)) was cloned and expressed in an Escherichia coli 2 Delta strain lacking the activities of alpha-ketoglutarate dehydrogenase (EC 1.2.4.2), isocitrate liase (EC 4.1.3.1), and isocitrate dehydrogenase kinase/phosphatase (EC 2.7.11.5). The 2 Delta strain could not grow in a minimal-salt/glucose/glycerol medium due to the blockage of TCA during succinate synthesis. The IDO activity in the 2 Delta strain was able to "shunt" destroyed TCA, thereby coupling l-isoleucine hydroxylation and cell growth. Using this strain, we performed the direct biotransformation of l-isoleucine into 4-HIL with an 82% yield.