Ajisaka Katsumi, Yuki Kaoru, Sato Kaori, Ishii Nozomi, Matsuo Ichiro, Kuji Ryo, Miyazaki Tatsuo, Furukawa Kiyoshi
a Department of Food Science , Niigata University of Pharmacy and Applied Life Sciences , Niigata , Japan.
b Department of Bioengineering , Nagaoka University of Technology , Nagaoka , Japan.
Biosci Biotechnol Biochem. 2016;80(1):128-34. doi: 10.1080/09168451.2015.1075863. Epub 2015 Aug 19.
Manα1 → 2Man, Manα1 → 3Man, Manα1 → 4Man, and Manα1 → 6Man were converted to the glycosylamine derivatives. Then, they were mixed with monobenzyl succinic acid to obtain their amide derivatives. After removing the benzyl group by hydrogenation, the succinylamide derivatives were coupled with the hydrazino groups on BlotGlyco™ beads in the presence of water-soluble carbodiimide. d-Mannobiose-linked beads were incubated with fluorescence-labeled Escherichia coli with type 1 fimbria, and the number of the fluorescent dots associated with the beads was counted in order to determine the binding preference among d-mannobiose isomers. The results showed that the bacteria bind strongly to Manα1 → 2Man1 → beads, Manα1 → 3Man1 → beads, Manα1 → 4Man1 → beads, and Manα1 → 6Man1 → beads, in order. In the presence of 0.1 M methyl α-d-mannopyranoside, most of the bacteria failed to bind to these beads. These results indicate that E. coli with type 1 fimbria binds to all types of d-mannobiose isomers but preferentially to Manα1 → 2Man disaccharide.
甘露糖α1→2甘露糖、甘露糖α1→3甘露糖、甘露糖α1→4甘露糖和甘露糖α1→6甘露糖被转化为糖基胺衍生物。然后，将它们与单苄基琥珀酸混合以获得其酰胺衍生物。通过氢化除去苄基后，在水溶性碳二亚胺存在下，琥珀酰胺衍生物与BlotGlyco™珠上的肼基偶联。将d-甘露二糖连接的珠子与荧光标记的1型菌毛大肠杆菌一起孵育，并对与珠子相关的荧光点数量进行计数，以确定d-甘露二糖异构体之间的结合偏好。结果表明，细菌与甘露糖α1→2甘露糖1→珠子、甘露糖α1→3甘露糖1→珠子、甘露糖α1→4甘露糖1→珠子和甘露糖α1→6甘露糖1→珠子的结合力依次增强。在0.1Mα-d-甘露吡喃糖苷存在下，大多数细菌无法与这些珠子结合。这些结果表明，1型菌毛大肠杆菌与所有类型的d-甘露二糖异构体结合，但优先与甘露糖α1→2甘露糖二糖结合。
