Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Asymmetric Sythesis of Unnatural L-Amino Acids Using Thermophilic Aromatic L-Amino Acid Transaminase

  • Cho, Byung-Kwan (School of Chemical and Biological Engineering and Institute of Molecular Biology and Genetics, Seoul National University) ;
  • Seo, Joo-Hyun (School of Chemical and Biological Engineering and Institute of Molecular Biology and Genetics, Seoul National University) ;
  • Kim, Ju-Han (School of Chemical and Biological Engineering and Institute of Molecular Biology and Genetics, Seoul National University) ;
  • Lee, Chang-Soo (Department of Chemical Engineering, Chungnam National University) ;
  • Kim, Byung-Gee (School of Chemical and Biological Engineering and Institute of Molecular Biology and Genetics, Seoul National University)
  • Published : 2006.08.30
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Abstract

Aromatic L-amino acid transaminase is an enzyme that is able to transfer the amino group from L-glutamate to unnatural aromatic ${\alpha}-keto$ acids to generate ${\alpha}-ketoglutarate$ and unnatural aromatic L-amino acids, respectively. Enrichment culture was used to isolate thermophilic Bacillus sp. T30 expressing this enzyme for use in the synthesis of unnatural L-amino acids. The asymmetric syntheses of L-homophenylalanine and L-phenylglycine resulted in conversion yields of >95% and >93% from 150 mM 2-oxo-4-phenylbutyrate and phenylglyoxylate, respectively, using L-glutamate as an amino donor at $60^{\circ}C$. Synthesized L-homophenylalanine and L-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 ${\alpha}-keto$ acid substrates is dependent on temperature, the solubility of the unnatural L-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.

Keywords

References

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