Metabolic engineering of Escherichia coli to produce (2S, 3R, 4S)-4-hydroxyisoleucine.
The stereo-specific L-isoleucine-4-hydroxylase (L-isoleucine dioxygenase (IDO)) was cloned and expressed in an Escherichia coli 2Δ strain lacking the activities of α-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 2Δ 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 2Δ 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.
Smirnov
Sergey V
SV
Ajinomoto-Genetika Research Institute, 1st Dorozhny Pr. 1, Moscow 117545, Russia. servasmi@mail.ru
Kodera
Tomohiro
T
Samsonova
Natalya N
NN
Kotlyarova
Veronika A
VA
Rushkevich
Natalya Yu
NY
Kivero
Alexander D
AD
Sokolov
Pavel M
PM
Hibi
Makoto
M
Ogawa
Jun
J
Shimizu
Sakayu
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D001711:Biotransformation; D003001:Cloning, Molecular; D049308:Dioxygenases / Q000378:metabolism; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial; D007532:Isoleucine / Q000031:analogs & derivatives* / Q000096:biosynthesis; D007655:Ketoglutarate Dehydrogenase Complex / Q000378:metabolism; D008969:Molecular Sequence Data
D007532:Isoleucine; C502516:(2S,3S,4R)-gamma-hydroxyisoleucine; D049308:Dioxygenases; D007655:Ketoglutarate Dehydrogenase Complex