Open AccessPost-translational Introduction of d -Alanine into Ribosomally Synthesized Peptides by the Dehydroalanine Reductase NpnJ
Author(s) -
Yang Xiao,
Wilfred A. van der Donk
Publication year - 2015
Publication title -
journal of the american chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.115
H-Index - 612
eISSN - 1520-5126
pISSN - 0002-7863
DOI - 10.1021/jacs.5b05207
Subject(s) - dehydroalanine , chemistry , amino acid , alanine , biochemistry , escherichia coli , stereochemistry , substrate (aquarium) , lantibiotics , enzyme , stereocenter , combinatorial chemistry , organic chemistry , catalysis , enantioselective synthesis , oceanography , antimicrobial , nisin , gene , geology
Ribosomally synthesized peptides are generally limited to L-amino acid building blocks. Given the advantageous properties of peptides containing D-amino acids such as stabilization of certain turns and against proteolytic degradation, methods to introduce D-stereocenters are valuable. Here we report the first in vitro reconstitution and characterization of a dehydrogenase that carries out the asymmetric reduction of dehydroalanine. NpnJA reduces dehydroalanine to D-Ala using NAPDH as cosubstrate. The enzyme displays high substrate tolerance allowing introduction of D-Ala into a range of non-native substrates. In addition to the in vitro reactions, we describe five examples of using Escherichia coli as biosynthetic host for D-alanine introduction into ribosomal peptides. A deuterium-label-based coupled-enzyme assay was used to rapidly determine the stereochemistry of the newly installed alanine.