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Appl. Environ. Microbiol., 12 1997, 4651-4656, Vol 63, No. 12
Copyright © 1997, American Society for Microbiology

Synthesis of optically active amino acids from alpha-keto acids with Escherichia coli cells expressing heterologous genes

A Galkin, L Kulakova, T Yoshimura, K Soda and N Esaki
Institute for Chemical Research, Kyoto University, Japan.

We describe a simple method for enzymatic synthesis of L and D amino acids from alpha-keto acids with Escherichia coli cells which express heterologous genes. L-amino acids were produced with thermostable L- amino acid dehydrogenase and formate dehydrogenase (FDH) from alpha- keto acids and ammonium formate with only an intracellular pool of NAD+ for the regeneration of NADH. We constructed plasmids containing, in addition to the FDH gene, the genes for amino acid dehydrogenases, including i.e., leucine dehydrogenase, alanine dehydrogenase, and phenylalanine dehydrogenase. L-Leucine, L-valine, L-norvaline, L- methionine, L-phenylalanine, and L-tyrosine were synthesized with the recombinant E. coli cells with high chemical yields (> 80%) and high optical yields (up to 100% enantiomeric excess). Stereospecific conversion of various alpha-keto acids to D amino acids was also examined with recombinant E. coli cells containing a plasmid coding for the four heterologous genes of the thermostable enzymes D-amino acid aminotransferase, alanine racemase, L-alanine dehydrogenase, and FDH. Optically pure D enantiomers of glutamate and leucine were obtained.

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