초록
<P><B>Abstract</B></P><P>Chiral amino acids are important intermediates for the pharmaceutical industry. We have developed a novel one‐pot enzymatic method for <SMALL>D</SMALL>‐amino acid synthesis by the dynamic kinetic resolution of <I>N</I>‐succinyl‐<SMALL>dl</SMALL>‐amino acids using <SMALL>D</SMALL>‐succinylase (DSA) and <I>N</I>‐succinylamino acid racemase (NSAR, EC 4.2.1.113). The DSA from <I>Cupriavidus</I> sp. P4‐10‐C, which hydrolyzes <I>N</I>‐succinyl‐<SMALL>D</SMALL>‐amino acids enantioselectively to their corresponding <SMALL>D</SMALL>‐amino acids, was identified for the first time by screening soil microorganisms. Subsequently, the DSA gene was cloned and overexpressed in <I>Escherichia coli</I>. DSA was shown to comprise two subunits with molecular masses of 26 kDa and 60 kDa. Additionally, the NSAR gene from <I>Geobacillus stearothermphilus</I> NCA1503, which racemizes <I>N</I>‐succinylamino acids, was also cloned and overexpressed in <I>E. coli</I>. The highly purified DSA and NSAR prepared from each recombinant <I>E. coli</I> were characterized and used for <SMALL>D</SMALL>‐amino acid synthesis. A one‐pot enzymatic method converted 100 mM <I>N</I>‐succinyl‐<SMALL>dl</SMALL>‐phenylalanine to <SMALL>D</SMALL>‐phenylalanine in 91.1% conversion with 86.7% <I>ee</I>. This novel enzymatic method may be useful for the industrial production of many <SMALL>D</SMALL>‐amino acids.</P>