The biosynthesis of colitose begins with ColE, a mannose-1-phosphate guanylyltransferase that catalyzes the addition of a GMP moiety to mannose, yielding GDP-mannose. In the next step, ColB, an NADP-dependent short-chain dehydrogenase-reductase enzyme, catalyzes the oxidation at C-4 and the removal of the hydroxyl group at C-6. The resulting product, GDP-4-keto-6-deoxymannose, then reacts with the PLP-dependent enzyme GDP-4-keto-6-deoxymannose-3-dehydratase (ColD), which removes the hydroxyl at C-3 in a manner similar to that of serine dehydratase. In the final step, the product of ColD, GDP-4-keto-3,6-dideoxymannose, reacts with ColC, which reduces the ketone functionality at C-4 back to an alcohol and inverts the configuration about C-5.[5]
The resulting product, GDP-L-colitose, is then incorporated into the O-antigen by glycosyltransferases and O-antigen processing proteins. Further reactions join the O-antigen to the core polysaccharide to form the full lipopolysaccharide.
GDP-4-keto-6-deoxymannose-3-dehydratase (ColD)
ColD is a PLP-dependent enzyme responsible for the removal of the C-3' hydroxyl group during the biosynthesis of GDP-colitose.[5] It is a product of the Wbdk or ColD genes in Escherichia coli O55 or Salmonella enterica, respectively, and is commonly referred to as ColD.[1]
Usage in biotechnology
Although the sugar is relatively rare, recent work with glycosyltransferases suggests that obscure sugars such as colitose can be incorporated into existing natural-product scaffolds, thereby constructing novel and potentially therapeutic compounds.[6]
References
^ abcSamuel G, Reeves P (November 2003). "Biosynthesis of O-antigens: genes and pathways involved in nucleotide sugar precursor synthesis and O-antigen assembly". Carbohydrate Research. 338 (23): 2503–19. doi:10.1016/j.carres.2003.07.009. PMID14670712.
^Muldoon J, Perepelov AV, Shashkov AS, et al. (June 2001). "Structure of a colitose-containing O-specific polysaccharide of the marine bacterium Pseudoalteromonas tetraodonis IAM 14160(T)". Carbohydrate Research. 333 (1): 41–6. doi:10.1016/S0008-6215(01)00121-5. PMID11423109.
^Luderitz O, Staub AM, Stirm S, Westphal O (1958). "[Colitose, 3-desoxy-L-fucose, a new sugar building block with immunospecific functions in the endotoxins, lipopolysaccharides, of some gram-negative bacteria.]". Biochemische Zeitschrift (in German). 330 (3): 193–7. PMID13546193.
^ abAlam J, Beyer N, Liu HW (December 2004). "Biosynthesis of colitose: expression, purification, and mechanistic characterization of GDP-4-keto-6-deoxy-D-mannose-3-dehydrase (ColD) and GDP-L-colitose synthase (ColC)". Biochemistry. 43 (51): 16450–60. doi:10.1021/bi0483763. PMID15610039.
^Zhang C, Griffith BR, Fu Q, et al. (September 2006). "Exploiting the reversibility of natural product glycosyltransferase-catalyzed reactions". Science. 313 (5791): 1291–4. doi:10.1126/science.1130028. PMID16946071. S2CID38072017.