title: Metabolic engineering of Escherichia coli to produce (2S, 3R, 4S)-4-hydroxyisoleucine.
abstract: 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.
authors.affiliation: Ajinomoto-Genetika Research Institute, 1st Dorozhny Pr. 1, Moscow 117545, Russia. servasmi@mail.ru
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Smirnov
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kodera
authors.forename: Natalya N
authors.initials: NN
authors.lastname: Samsonova
authors.forename: Veronika A
authors.initials: VA
authors.lastname: Kotlyarova
authors.forename: Natalya Yu
authors.initials: NY
authors.lastname: Rushkevich
authors.forename: Alexander D
authors.initials: AD
authors.lastname: Kivero
authors.forename: Pavel M
authors.initials: PM
authors.lastname: Sokolov
authors.forename: Makoto
authors.initials: M
authors.lastname: Hibi
authors.forename: Jun
authors.initials: J
authors.lastname: Ogawa
authors.forename: Sakayu
authors.initials: S
authors.lastname: Shimizu
mesh_terms: 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
chemical_list: D007532:Isoleucine; C502516:(2S,3S,4R)-gamma-hydroxyisoleucine; D049308:Dioxygenases; D007655:Ketoglutarate Dehydrogenase Complex