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Asymmetric synthesis of unnaturall-amino acids using thermophilic aromaticl-amino acid transaminase

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Abstract

Aromaticl-amino acid transaminase is an enzyme that is able to transfer the amino group froml-glutamate to unnatural aromatic α-keto acids to generate α-ketoglutarate and unnatural aromaticl-amino acids, respectively. Enrichment culture was used to isolate thermophilicBacillus sp. T30 expressing this enzyme for use in the synthesis of unnaturall-amino acids. The asymmetric syntheses ofl-homophenylalanine andl-phenylglycine resulted in conversion yields of >95% and >93% from 150 mM 2-oxo-4-phenylbutyrate and phenylglyoxylate, respectively, usingl-glutamate as an amino donor at 60°C. Synthesizedl-homophenylalanine andl-phenylglycine were optically pure (>99% enantiomeric excess) and continuously pre-cipitated in the reaction solution due to their low solubility at the given reaction pH. While the solubility of the α-keto acid substrates is dependent on temperature, the solubility of the unnaturall-amino acid products is dependent on the reaction pH. As the solubility difference between substrate and product at the given reaction pH is therefore larger at higher temperature, the thermophilic transaminase was successfully used to shift the reaction equilibrium toward rapid product formation.

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Author notes

  1. Byung-Kwan Cho

    Present address: Department of Bioengineering, University of California, San Diego, 92093, La Jolla, CA, USA

  2. Juhan Kim

    Present address: Department of Molecular Cellular and Developmental Biology, University of Colorado, 80309, Boulder, CO, USA

Authors and Affiliations

  1. School of Chemical and Biological Engineering and Institute of Molecular Biology and Genetics, Seoul National University, 151-742, Seoul, Korea

    Byung-Kwan Cho, Joo-Hyun Seo, Juhan Kim & Byung-Gee Kim

  2. Department of Chemical Engineering, Chungnam National University, 305-764, Daejeon, Korea

    Chang-Soo Lee

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  1. Byung-Kwan Cho
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  2. Joo-Hyun Seo
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  5. Byung-Gee Kim
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Correspondence to Byung-Gee Kim.

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Cho, BK., Seo, JH., Kim, J. et al. Asymmetric synthesis of unnaturall-amino acids using thermophilic aromaticl-amino acid transaminase. Biotechnol. Bioprocess Eng. 11, 299–305 (2006). https://doi.org/10.1007/BF03026244

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  • DOI: https://doi.org/10.1007/BF03026244

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