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Metabolic engineering of Escherichia coli to produce (2S, 3R, 4S)-4-hydroxyisoleucine

  • Applied Genetics and Molecular Biotechnology
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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.

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Authors and Affiliations

  1. Ajinomoto-Genetika Research Institute, 1st Dorozhny pr. 1, Moscow, 117545, Russia

    Sergey V. Smirnov, Natalya N. Samsonova, Veronika А. Kotlyarovа, Natalya Yu Rushkevich, Аlexander D. Kivero & Pavel M. Sokolov

  2. Institute of Life Sciences, Ajinomoto Co., Inc., Suzuki-cho, Kawasaki-ku, Kawasaki-shi, 210-8681, Japan

    Tomohiro Kodera

  3. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, 8 Sakyo-ku, Kyoto, 606-8502, Japan

    Makoto Hibi, Jun Ogawa & Sakayu Shimizu

Authors
  1. Sergey V. Smirnov
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  2. Tomohiro Kodera
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  3. Natalya N. Samsonova
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  4. Veronika А. Kotlyarovа
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  5. Natalya Yu Rushkevich
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  6. Аlexander D. Kivero
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  7. Pavel M. Sokolov
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  8. Makoto Hibi
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  9. Jun Ogawa
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  10. Sakayu Shimizu
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Correspondence to Sergey V. Smirnov.

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Smirnov, S.V., Kodera, T., Samsonova, N.N. et al. Metabolic engineering of Escherichia coli to produce (2S, 3R, 4S)-4-hydroxyisoleucine. Appl Microbiol Biotechnol 88, 719–726 (2010). https://doi.org/10.1007/s00253-010-2772-3

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  • DOI: https://doi.org/10.1007/s00253-010-2772-3

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