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- Title
Metabolic engineering of Escherichia coli to produce (2S, 3R, 4S)-4-hydroxyisoleucine.
- Authors
Smirnov, Sergey V.; Kodera, Tomohiro; Samsonova, Natalya N.; Kotlyarova, Veronika A.; Rushkevich, Natalya Yu; Kivero, Alexander D.; Sokolov, Pavel M.; Hibi, Makoto; Ogawa, Jun; Shimizu, Sakayu
- 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.
- Subjects
ESCHERICHIA coli physiology; HYDROXYLATION; DEHYDROGENASES; GLYCERIN; BIOTRANSFORMATION (Metabolism); DRUG activation; TREATMENT of diabetes
- Publication
Applied Microbiology & Biotechnology, 2010, Vol 88, Issue 3, p719
- ISSN
0175-7598
- Publication type
Article
- DOI
10.1007/s00253-010-2772-3