Bacterial coupling

The first prominent example of large-scale oligosaccharide production by bacterial coupling was described by Koizumi et al. (Scheme 5.7) [40]. Three engineered bacterial strains were combined for the production of globotriose (13) including regeneration of UDP-Gal. 

A very efficient and economical whole cell-based bacterial coupling method for the synthesis of many sugar nucleotide donors and oligosaccharides has been developed (188). In the... 

Endo T, Koizumi S, Tabata K, Kakita S, Ozaki A. Large-scale production of W-acetyllactosamine through bacterial coupling. 90. Carbohydr. Res. 1999 316 179-183. 

Our new ability to obtain a highly purified preparation of a bacterial coupling factor PPase, in combination with our recently obtained capacity to clone any part of the R. rubrum genome, may open the road to a detailed study of the molecular properties of the enzyme. The techniques now available for directed mutations of bacterial genomes may give us possibilities to alter the dynamic properties of the enzyme in ways that should provide added insight into the mechanism of formation and utilization of inorganic pyrophosphate. 

Koizumi S, Endo T, Tabata K, Nagano H, Ohnishi J, Ozaki A. Large-scale production of GDP-fucose and Lewis X by bacterial coupling. J Ind Microbiol Biotechnol 2000 25 213-217. 

A system of oligosaccharide production by the combination of a bacterial glycosyltransferase and sugar nucleotide production was proposed on the basis that sugar nucleotides could be synthesized by bacterial coupling. 

Fig. 6 Production of 3 sialyllactose by bacterial coupling (36). (Reproduced with permission from reference 36. Copyright 2000 Springer- Verlag.)...

By coupling of the sugar nucleotides production system with bacterial glycosyltransferase, an oligosaccharide production system that could be applied to the industrial manufacture was successfully constructed. Oligosaccharides production by bacterial coupling is suitable for large-scale synthesis. 

Scheme 8 Kyowa Hakko s technology through multiple bacterial coupling...

At present, conditions are being developed to improve the separation between the peaks containing the a and 3 subunits. Since the conditions used for subunit separation are relatively mild, and since the a and 3 subunits are soluble in the absence of urea, it may be possible to study the function of these subunits either in reconstitution experiments or perhaps alone as has been done in the case of the bacterial coupling factor (Dunn, Futai, 1980) and in the case of the 6 subunit of CFi (Andreo et al., 1982). 

---