Division of Life Science, Graduate School of Science and Engineering, Saitama University, 255 Shimo-okubo, Sakura-ku, Saitama, 338-8570, Japan.
R&D-Biological Science Research, Kao Corporation, 2606 Akabane, Ichikai-Machi, Haga-Gun, Tochigi, 321-3497, Japan.
Carbohydr Res. 2019 Nov 1;485:107816. doi: 10.1016/j.carres.2019.107816. Epub 2019 Sep 13.
Arabinogalactans (AGs) and arabinogalactan-proteins (AGPs) were partially purified from an extract of fruits of the European pear (Pyrus communis L.) by DEAE-cellulose ion-exchange and Sepharose 6B gel-filtration chromatography. Among 7 AG(P)-containing fractions, a neutral AGP (SE-1) was confirmed to be highly purified (M 67,000) and rich in L-Ara and Gal; this fraction included a small amount (2.6%, w/w) of protein and showed the highest reactivity forming precipitate with β-Glc Yariv reagent among the 7 fractions, the intensity of which was comparable to that of gum arabic, a standard AGP. Another accompanying minor low-M neutral AGP (SE-2; M approx. 7200) still contained other polysaccharide (starch fragments) and did not show Yariv reactivity. The carbohydrate moieties of SE-1 consisted of consecutive (1 → 3)-linked β-galactosyl backbone chains substituted with side chains of (1 → 6)-linked β-galactosyl residues at O-6, to which mainly single α-l-arabinofuranosyl residues were attached through O-3. This structural feature was also observed for SE-2. Successive digestion of SE-1 with α-l-arabinofuranosidase and exo-β-(1 → 3)-galactanase with the aid of endo-β-(1 → 3)-galactanase released most (more than 98%, w/w) of the carbohydrate moieties as low-M fragments. These consisted of free L-Ara and Gal, and a series of β-(1 → 6)-galactooligosaccharides with degree of polymerization (dp) up to at least 17, indicative of attachment of (1 → 6)-linked β-galactosyl side chains of varying length along the (1 → 3)-linked β-galactosyl backbone chains.
阿拉伯半乳聚糖(AGs)和阿拉伯半乳聚糖蛋白(AGPs)从欧洲梨(Pyrus communis L.)果实提取物中通过 DEAE-纤维素离子交换和 Sepharose 6B 凝胶过滤层析部分纯化。在 7 个含有 AG(P)的级分中,确认中性 AGP(SE-1)高度纯化(M 67,000)且富含 L-Ara 和 Gal;该级分含有少量(2.6%,w/w)的蛋白质,与 7 个级分中β-Glc Yariv 试剂形成沉淀的反应性最强,其强度与标准 AGP 阿拉伯胶相当。另一个伴随的次要低分子量中性 AGP(SE-2;M 约 7200)仍含有其他多糖(淀粉片段),并且不显示 Yariv 反应性。SE-1 的碳水化合物部分由连续(1 → 3)-连接的β-半乳糖基主链组成,侧链由 O-6 上的(1 → 6)-连接的β-半乳糖基残基取代,主要通过 O-3 连接单α-L-阿拉伯呋喃糖基残基。SE-2 也观察到这种结构特征。SE-1 用α-L-阿拉伯呋喃糖苷酶和外切-β-(1 → 3)-半乳糖苷酶连续消化,在内切-β-(1 → 3)-半乳糖苷酶的帮助下,将大部分(超过 98%,w/w)碳水化合物部分释放为低分子量片段。这些由游离的 L-Ara 和 Gal 以及一系列β-(1 → 6)-半乳糖低聚糖组成,聚合度(dp)高达至少 17,表明在(1 → 3)-连接的β-半乳糖基主链上具有不同长度的(1 → 6)-连接的β-半乳糖基侧链的连接。