Production of (R)-3-amino-3-phenylpropionic acid and (S)-3-amino-3-phenylpropionic acid from (R,S)-N-acetyl-3-amino-3-phenylpropionic acid using microorganisms having enantiomer-specific amidohydrolyzing activity.
(R)-3-Amino-3-phenylpropionic acid ((R)-beta-Phe) and (S)-3-amino-3-phenylpropionic acid ((S)-beta-Phe) are key compounds on account of their use as intermediates in synthesizing pharmaceuticals. Enantiomerically pure non-natural amino acids are generally prepared by enzymatic resolution of the racemic N-acetyl form, but despite the intense efforts this method could not be used for preparing enantiomerically pure beta-Phe, because the effective enzyme had not been found. Therefore, screening for microorganisms capable of amidohydrolyzing (R,S)-N-acetyl-3-amino-3-phenylpropionic acid ((R,S)-N-Ac-beta-Phe) in an enantiomer-specific manner was performed. A microorganism having (R)-enantiomer-specific amidohydrolyzing activity and another having both (R)-enantiomer- and (S)-enantiomer-specific amidohydrolyzing activities were obtained from soil samples. Using 16S rDNA analysis, the former organism was identified as Variovorax sp., and the latter as Burkholderia sp. Using these organisms, enantiomerically pure (R)-beta-Phe (>99.5% ee) and (S)-beta-Phe (>99.5% ee) with a high molar conversion yield (67%-96%) were obtained from the racemic substrate.
Kawasaki
Hisashi
H
Department of Environmental Materials Science, Tokyo Denki University, Tokyo, Japan.
Koyama
Koutaro
K
Kurokawa
Sachio
S
Watanabe
Kunihiko
K
Nakazawa
Masakazu
M
Izawa
Kunisuke
K
Nakamatsu
Tsuyoshi
T
D002474:Cell-Free System; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry; D012988:Soil Microbiology; D013237:Stereoisomerism; D013379:Substrate Specificity; D013696:Temperature
C064718:3-amino-3-phenylpropionic acid; D010649:Phenylalanine