Premium
Identification of the GDP‐ N ‐acetyl‐ d ‐perosamine producing enzymes from Escherichia coli O157:H7
Author(s) -
Albermann Christoph,
Beuttler Holger
Publication year - 2008
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2008.01.005
Subject(s) - escherichia coli , transamination , enzyme , chemistry , biosynthesis , biochemistry , glycosyltransferase , recombinant dna , mass spectrometry , stereochemistry , chromatography , gene
GDP‐ N ‐acetyl‐ d ‐perosamine is a precursor of the LPS‐O‐antigen biosynthesis in Escherichia coli O157:H7. Like other GDP‐6‐deoxyhexoses, GDP‐ N ‐acetyl‐ d ‐perosamine is supposed to be synthesized via GDP‐4‐keto‐6‐deoxy‐ d ‐mannose, followed by a transamination‐ and an acetylation‐reaction catalyzed by PerA and PerB. In this study, we have overproduced and purified PerA and PerB from E. coli O157:H7 in E. coli BL21. The recombinant proteins were partly characterized and the final product of the reaction catalyzed by PerB was shown to be GDP‐ N ‐acetyl‐ d ‐perosamine by chromatography, mass spectrometry, and 1 H‐NMR. The functional expression of PerB provides another enzymatically defined pathway for the synthesis of GDP‐deoxyhexoses, which is needed to further study the corresponding glycosyltransferases in vitro.