An Fe(II)/α-ketoglutarate-dependent dioxygenase, SadA, was obtained from Burkholderia ambifaria AMMD and heterologously expressed in Escherichia coli. Purified recombinant SadA had catalytic activity towards several N-substituted l-amino acids, which was especially strong with N-succinyl l-leucine. With the NMR and LC-MS analysis, SadA converted N-succinyl l-leucine into N-succinyl l-threo-β-hydroxyleucine with >99% diastereoselectivity. SadA is the first enzyme catalysing β-hydroxylation of aliphatic amino acid-related substances and a potent biocatalyst for the preparation of optically active β-hydroxy amino acids.
Keywords: Burkholderia ambifaria; Fe(II)/α‐ketoglutarate‐dependent dioxygenase; N‐substituted l‐amino acid β‐hydroxylase.
© 2012 The Society for Applied Microbiology.