BioChem - DOS

An Improved Racemase/Acylase Biotransformation for the Preparation of Enantiomerically Pure Amino Acids

Journal of the American Chemical Society

Baxter, Scott; Royer, Sylvain; Grogan, Gideon; Brown, Fraser; Holt-Tiffin, Karen E.; Taylor, Ian N.; Fotheringham, Ian G.; Campopiano, Dominic J.

<P>Using directed evolution, a variant <I>N</I>-acetyl amino acid racemase (NAAAR G291D/F323Y) has been developed with up to 6-fold higher activity than the wild-type on a range of <I>N</I>-acetylated amino acids. The variant has been coupled with an enantiospecific acylase to give a preparative scale dynamic kinetic resolution which allows 98% conversion of <I>N</I>-acetyl-<SMALL>dl</SMALL>-allylglycine into <SMALL>d</SMALL>-allylglycine in 18 h at high substrate concentrations (50 g L<SUP>&ndash;1</SUP>). This is the first example of NAAAR operating under conditions which would allow it to be successfully used on an industrial scale for the production of enantiomerically pure &alpha;-amino acids. X-ray crystal analysis of the improved NAAAR variant allowed a comparison with the wild-type enzyme. We postulate that a network of novel interactions that result from the introduction of the two side chains is the source of improved catalytic performance.</P><P><B>Graphic Abstract</B><BR><IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jacsat/2012/jacsat.2012.134.issue-47/ja305438y/production/images/medium/ja-2012-05438y_0005.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/ja305438y'>ACS Electronic Supporting Info</A></P>

2012

American Chemical Society

0002-7863

1520-5126

134

47

pp.19310-19313

10.1021/ja305438y

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART65056366

https://www.pure.ed.ac.uk/ws/files/10175419/An_Improved_RacemaseAcylase_Biotransformation_for_the_Preparation_of_Enantiomerically_Pure_Amino_Acids.pdf