Reuhs B L, Carlson R W, Kim J S
Complex Carbohydrate Research Center, University of Georgia, Athens 30602-4712.
J Bacteriol. 1993 Jun;175(11):3570-80. doi: 10.1128/jb.175.11.3570-3580.1993.
The polysaccharide components from cultured cells of Rhizobium fredii USDA205 and Rhizobium meliloti AK631 were extracted with hot phenol-water and separated by repetitive gel filtration chromatography. Polyacrylamide gel electrophoresis, nuclear magnetic resonance spectrometry, and gas chromatography analyses showed that both of these bacterial species produce unique polysaccharides that contain a high proportion of 3-deoxy-D-manno-2-octulosonic acid (Kdo). These polysaccharides, which constituted a major portion of the extracted carbohydrate, are not excreted into the growth media (i.e., they are not extracellular polysaccharides) and are structurally distinct from the lipopolysaccharides. The primary structure of the preponderant polysaccharide from R. fredii USDA205 was determined by high-performance anion-exchange liquid chromatography, nuclear magnetic resonance spectrometry, fast atom bombardment-mass spectrometry, and gas chromatography-mass spectrometry; it consists of repeating units of [-->3)-alpha-D-Galp-(1-->5)-beta-D-Kdop-(2-->]n. This molecule is structurally analogous to the constituents of one subgroup of K antigens (capsular polysaccharides) produced by Escherichia coli. Polysaccharides of this type have not previously been identified as components of rhizobial cells. The Kdo-containing polysaccharide from R. meliloti, which has not been completely characterized, appears to be structurally related to that of R. fredii.
用热酚水法提取费氏中华根瘤菌USDA205和苜蓿根瘤菌AK631培养细胞中的多糖成分,并通过反复凝胶过滤色谱法进行分离。聚丙烯酰胺凝胶电泳、核磁共振光谱和气相色谱分析表明,这两种细菌都能产生独特的多糖,其中含有高比例的3-脱氧-D-甘露-2-辛酮糖酸(Kdo)。这些多糖构成了提取碳水化合物的主要部分,它们不会分泌到生长培养基中(即它们不是胞外多糖),并且结构上与脂多糖不同。通过高效阴离子交换液相色谱、核磁共振光谱、快原子轰击质谱和气相色谱-质谱法确定了费氏中华根瘤菌USDA205中优势多糖的一级结构;它由[-->3)-α-D-吡喃半乳糖-(1-->5)-β-D-Kdo-(2-->]n的重复单元组成。该分子在结构上类似于大肠杆菌产生的K抗原(荚膜多糖)一个亚组的成分。此前尚未将这种类型的多糖鉴定为根瘤菌细胞的成分。苜蓿根瘤菌中含Kdo的多糖尚未完全表征,但其结构似乎与费氏中华根瘤菌的多糖有关。