Engineered phenylalanine dehydrogenase in organic solvents: homogeneous and biphasic enzymatic reactions

Org Biomol Chem. 2005 Dec 21;3(24):4316-20. doi: 10.1039/b510816k. Epub 2005 Nov 14.

Abstract

The use of engineered phenylalanine dehydrogenase N145A supported on Celite for the reductive amination of phenylpyruvic acid in homogeneous and biphasic aqueous-organic solvents is reported. The results indicate that the immobilised biocatalyst is remarkably robust, even in the presence of high concentrations of polar or non-polar organic solvents such as acetone, methanol, n-hexane, toluene and methylene chloride. Cofactor regeneration with alcohol dehydrogenase from Saccharomyces cerevisiae and ethanol was successfully explored. Application to the non-natural poorly water-soluble 2-oxo acid p-NO(2)-phenylpyruvic acid was successfully performed, resulting in the biocatalytic synthesis of p-NO(2)-phenylalanine. In all cases 100% stereoselectivity for the production of the amino acid was retained.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Oxidoreductases / metabolism*
  • Chemistry, Organic
  • Molecular Structure
  • Nitrogen Dioxide / chemistry
  • Organic Chemistry Phenomena
  • Phenylalanine / chemistry
  • Phenylalanine / metabolism
  • Phenylpyruvic Acids / chemistry
  • Phenylpyruvic Acids / metabolism
  • Protein Engineering
  • Solvents

Substances

  • Phenylpyruvic Acids
  • Solvents
  • Phenylalanine
  • Amino Acid Oxidoreductases
  • phenylalanine oxidase
  • Nitrogen Dioxide
  • phenylpyruvic acid