Orgeret C, Seillier E, Gautier C, Defaye J, Driguez H
Centre de Recherches sur les Macromolécules Végétales, C.N.R.S., Grenoble, France.
Carbohydr Res. 1992 Feb 7;224:29-40. doi: 10.1016/0008-6215(92)84090-f.
4-Aminophenyl 1,4-dithio-beta-cellobioside (6) was obtained by treatment of methyl 2,3,6-tri-O-benzoyl-4-O-triflyl-alpha-D-galactopyranoside with the sodium salt of 1-thio-beta-D-glucopyranose, followed by acetolysis and glycosylation of the corresponding bromide with 4-aminobenzenethiol and subsequent deacylation. A similar synthesis starting with the 1-thiolate of 1,4-dithio-beta-cellobiose led to the trisaccharide 4-aminophenyl 1,4,4'-trithiocellotrioside (16). The 4-acetamidophenyl di- and tri-thiocellooligosaccharides were found to be excellent competitive inhibitors of the hydrolysis of 4-methylumbelliferyl beta-lactoside with respective Ki values of 25 and 6.5 mM. The two 4-aminophenyl oligosaccharides 6 and 16 were coupled to CH-Sepharose 4B, and the affinity gels were used for the purification of cellobiohydrolases from a crude commercial cellulolytic extract of T. reesei. Cellobiohydrolases I or II were selectively desorbed from gels bearing ligands 6 and 16.
4-氨基苯基 1,4-二硫代-β-纤维二糖苷（6）的制备方法为：用 1-硫代-β-D-吡喃葡萄糖钠盐处理 2,3,6-三-O-苯甲酰基-4-O-三氟甲磺酰基-α-D-吡喃半乳糖苷甲酯，接着进行乙酰解，再用 4-氨基苯硫酚对相应的溴化物进行糖基化，随后脱酰基。以 1,4-二硫代-β-纤维二糖的 1-硫醇盐为起始原料进行类似合成，得到三糖 4-氨基苯基 1,4,4'-三硫代纤维三糖苷（16）。发现 4-乙酰氨基苯基二硫代和三硫代纤维寡糖是 4-甲基伞形酮基-β-乳糖苷水解的优秀竞争性抑制剂，其 Ki 值分别为 25 和 6.5 mM。将两种 4-氨基苯基寡糖 6 和 16 偶联到 CH-琼脂糖 4B 上，所得亲和凝胶用于从里氏木霉的粗制商业纤维素分解提取物中纯化纤维二糖水解酶。纤维二糖水解酶 I 或 II 可从带有配体 6 和 16 的凝胶上选择性解吸下来。
