Talaga P, Stahl B, Wieruszeski J M, Hillenkamp F, Tsuyumu S, Lippens G, Bohin J P
Laboratoire de Chimie Biologique, Centre National de la Recherche Scientifique UMR 11, Université des Sciences et Technologies de Lille, France.
J Bacteriol. 1996 Apr;178(8):2263-71. doi: 10.1128/jb.178.8.2263-2271.1996.
The cell-associated glucans produced by Burkholderia solanacearum and Xanthomonas campestris pv. citri were isolated by trichloroacetic acid treatment and gel permeation chromatography. The compounds obtained were characterized by compositional analysis, matrix-assisted laser desorption ionization mass spectrometry, and high-performance anion-exchange chromatography. B. solanacearum synthesizes only a neutral cyclic glucan containing 13 glucose residues, and X. campestris pv. citri synthesizes a neutral cyclic glucan containing 16 glucose residues. The two glucans were further purified by high-performance anion-exchange chromatography. Methylation analysis revealed that these glucans are linked by 1,2-glycosidic bonds and one 1,6-glycosidic bond. Our 600-MHz homonuclear and 1H-13C heteronuclear nuclear magnetic resonance experiments revealed the presence of a single alpha-1,6-glycosidic linkage, whereas all other glucose residues are beta-1,2 linked. The presence of this single alpha-1,6 linkage, however, induces such structural constraints in these cyclic glucans that all individual glucose residues could be distinguished. The different anomeric proton signals allowed complete sequence-specific assignment of both glucans. The structural characteristics of these glucans contrast with those of the previously described osmoregulated periplasmic glucans.
通过三氯乙酸处理和凝胶渗透色谱法分离了由青枯雷尔氏菌和柑橘溃疡病菌产生的细胞相关葡聚糖。对所得化合物进行了成分分析、基质辅助激光解吸电离质谱分析和高效阴离子交换色谱分析。青枯雷尔氏菌仅合成一种含有13个葡萄糖残基的中性环状葡聚糖,而柑橘溃疡病菌合成一种含有16个葡萄糖残基的中性环状葡聚糖。这两种葡聚糖通过高效阴离子交换色谱法进一步纯化。甲基化分析表明,这些葡聚糖通过1,2-糖苷键和一个1,6-糖苷键连接。我们的600兆赫兹同核和1H-13C异核核磁共振实验表明存在一个单一的α-1,6-糖苷键,而所有其他葡萄糖残基均为β-1,2连接。然而,这个单一的α-1,6键的存在在这些环状葡聚糖中诱导了这样的结构限制,以至于所有单个葡萄糖残基都可以被区分。不同的异头质子信号使得两种葡聚糖都能进行完整的序列特异性归属。这些葡聚糖的结构特征与先前描述的渗透调节周质葡聚糖的结构特征形成对比。