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1. (Article ID: 1273)
 
Suzuki N, Nakano Y, Yoshida Y, Nakao H, Yamashita Y, Koga T
Genetic analysis of the gene cluster for the synthesis of serotype a-specific polysaccharide antigen in Actinobacillus actinomycetemcomitans
Biochimica et Biophysica Acta: Gene Structure and Expression 1517(1) (2000) 135-138
 

The serotype a-specific polysaccharide antigen (SPA) of Actinobacillus actinomycetemcomitans consists of 6-deoxy-D-talose. A gene cluster associated with the biosynthesis of SPA was cloned and sequenced from the chromosomal DNA of A. actinomycetemcomitans SUNYaB 75 (serotype a). This cluster consisted of 14 open reading frames. Insertional inactivation of eight genes in this cluster resulted in loss of the ability of A. actinomycetemcomitans SUNYaB 75 cells to produce the polysaccharide. A protein database search revealed that the 11 sequential genes containing these eight genes were not found in SPA-associated gene clusters of the other serotypes of A. actinomycetemcomitans. These results suggest that the gene cluster is unique to serotype a and is essential to the synthesis of the SPA.

genetic, synthesis, antigen, gene, polysaccharide, serotype, analysis, cluster, gene cluster, Actinobacillus, Actinobacillus actinomycetemcomitans, polysaccharide antigen, serotype antigen

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2. (Article ID: 1274)
 
Suzuki N, Nakano Y, Yoshida Y, Nezu T, Terada Y, Yamashita Y, Koga T
Guanosine diphosphate-4-keto-6-deoxy-d-mannose reductase in the pathway for the synthesis of GDP-6-deoxy-D-talose in Actinobacillus actinomycetemcomitans
European Journal of Biochemistry 269(23) (2002) 5963-5971
 

The serotype a-specific polysaccharide antigen of Actinobacillus actinomycetemcomitans is an unusual sugar, 6-deoxy-d-talose. Guanosine diphosphate (GDP)-6-deoxy-d-talose is the activated sugar nucleotide form of 6-deoxy-d-talose, which has been identified as a constituent of only a few microbial polysaccharides. In this paper, we identify two genes encoding GDP-6-deoxy-d-talose synthetic enzymes, GDP-α-D-mannose 4,6-dehydratase and GDP-4-keto-6-deoxy-d-mannose reductase, in the gene cluster required for the biosynthesis of serotype a-specific polysaccharide antigen from A. actinomycetemcomitans SUNYaB 75. Both gene products were produced and purified from Escherichia coli transformed with plasmids containing these genes. Their enzymatic reactants were analysed by reversed-phase HPLC (RP-HPLC). The sugar nucleotide produced from GDP-α-D-mannose by these enzymes was purified by RP-HPLC and identified by electrospray ionization-MS, 1H nuclear magnetic resonance, and GC/MS. The results indicated that GDP-6- deoxy-d-talose is produced from GDP-α-D-mannose. This paper is the first report on the GDP-6-deoxy-d-talose biosynthetic pathway and the role of GDP-4-keto-6-deoxy-d-mannose reductase in the synthesis of GDP- 6-deoxy-d-talose

NMR, biosynthesis, synthesis, antigen, biosynthetic, gene, role, polysaccharide, serotype, form, Escherichia, Escherichia coli, cluster, gene cluster, electrospray, sugar, polysaccharides, enzymatic, nuclear, nuclear magnetic resonance, plasmid, resonance, enzyme, purified, Enzymes, pathway, Japan, Synthetic, Mannose, Actinobacillus, Actinobacillus actinomycetemcomitans, polysaccharide antigen, activated, 6-deoxy-D-talose, HPLC, Plasmids, oral, sugar nucleotide, 6-deoxytalose, microbial polysaccharides, serotype-specific antigen

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