Engineered phenylalanine dehydrogenase in organic solvents: homogeneous and biphasic enzymatic reactions.
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.
Cainelli
Gianfranco
G
Department of Chemistry "G. Ciamician", University of Bologna, Via Selmi 2, I-40126 Bologna, Italy. gianfranco.cainelli@unibo.it
Engel
Paul C
PC
Galletti
Paola
P
Giacomini
Daria
D
Gualandi
Andrea
A
Paradisi
Francesca
F
D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D002625:Chemistry, Organic; D015394:Molecular Structure; D009585:Nitrogen Dioxide / Q000737:chemistry; D055601:Organic Chemistry Phenomena; D010649:Phenylalanine / Q000737:chemistry / Q000378:metabolism; D010667:Phenylpyruvic Acids / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering; D012997:Solvents
D010667:Phenylpyruvic Acids; D012997:Solvents; D010649:Phenylalanine; D000594:Amino Acid Oxidoreductases; C022342:phenylalanine oxidase; D009585:Nitrogen Dioxide; C031606:phenylpyruvic acid