کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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1388760 | 982815 | 2008 | 12 صفحه PDF | دانلود رایگان |
Alloiococcus otitidis is a recently discovered Gram-positive bacterium that has been linked with otitis media (middle ear infections). In this study, we describe the structure of a novel capsular polysaccharide (PS) expressed by the type-strain of A. otitidis, ATCC 51267, and the synthesis of a glycoconjugate composed of the capsule PS and bovine serum albumin (BSA). The capsule PS of A. otitidis type-strain was determined to be a repeating trisaccharide composed of 3-substituted N-acetyl-d-glucosamine (GlcpNAc), 6-substituted N-acetyl-d-galactosamine (GalpNAc), and 4-substituted d-glucuronic acid (GlcpA), of which the majority was amidically decorated with l-glutamic acid (Glu): {→6)-β-GalpNAc-(1→4)-[Glup→6]-β-GlcpA-(1→3)-β-GlcpNAc-(1}n. Monomeric analysis performed on other A. otitidis strains revealed that similar components were variably expressed, but Glu appeared to be a regular constituent in all the strains examined. Due to the suitable presence of GlcpA and Glu, our approach for glycoconjugate synthesis employed a carbodiimide-based strategy with activation of available carboxyl groups by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC), which afforded direct coupling between the capsule PS and BSA. Analysis by mass spectrometry indicated that this A. otitidis capsule PS–BSA conjugate was composed of BSA units that carried up to seven capsule PSs. This work represents the first report in the literature describing an A. otitidis cell-surface carbohydrate and the synthesis of a glycoconjugate preparation thereof. Presently, we are formulating plans to immunologically evaluate this A. otitidis glycoconjugate vaccine in animals.
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Journal: Carbohydrate Research - Volume 343, Issue 6, 5 May 2008, Pages 1079–1090