Open AccessBiosynthesis of GDP-d -glycero-α-d -manno-heptose for the Capsular Polysaccharide of Campylobacter jejuni
Author(s) -
Jamison P. Huddleston,
Frank M. Raushel
Publication year - 2019
Publication title -
biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.43
H-Index - 253
eISSN - 1520-4995
pISSN - 0006-2960
DOI - 10.1021/acs.biochem.9b00548
Subject(s) - heptose , campylobacter jejuni , biosynthesis , enzyme , biochemistry , stereochemistry , chemistry , polysaccharide , biology , bacteria , mutant , gene , genetics
The capsular polysaccharide (CPS) structure of Campylobacter jejuni contributes to its robust fitness. Many strains contain heptose moieties in their CPS units. The precursor heptose is GDP-d- glycero -α-d- manno -heptose; modifications to the stereochemistry at C3-C6 as well as additions of methyl and phosphoramidate groups lend to the hypervariability of the C. jejuni CPS structures. Synthesis of GDP-d- glycero -α-d- manno -heptose has been described previously, but using enzymes from Aneurinibacillus thermoaerophilus DSM 10155. Here we describe the complete synthesis of GDP-d- glycero -α-d- manno -heptose using enzymes from C. jejuni NTCC 11168: Cj1152 and Cj1423-Cj1425. Our results yield kinetic parameters for these enzymes and outline a successful strategy for milligram-gram scale synthesis of GDP-d- glycero -α-d- manno -heptose. This achievement is critical for the characterization of other carbohydrate tailoring enzymes, which are expected to utilize GDP-d- glycero -α-d- manno -heptose for the biosynthesis of more complex carbohydrates in the CPS of C. jejuni .