葡萄糖基-β-环糊精的“一锅法”合成及其分离纯化
“One-pot” Synthesis, Separation and Puificatin of Glucosyl-β-cyclodextrin
葡萄糖基-β-环糊精（G1-β-CD）具有比母体β-环糊精（β-CD）更好的溶解性及生物相容性，其在食品、医药、农业及化工等领域的应用前景广泛，但因其制备及分离困难而导致目前国内生产研究者甚少。因此本文对G1-β-CD的合成及分离进行了研究，利用普鲁兰酶逆向合成作用，将麦芽糖（G2）接枝到β-CD上生成麦芽糖基-β-环糊精（G2-β-CD），然后不改变体系任何反应条件，加入糖化酶水解得到G1-β-CD，再用凝胶层析柱分离得到G1-β-CD纯品。结果表明普鲁兰酶逆向合成的最优条件为底物G2和β-CD摩尔比为8：1，加酶量360U/g β-CD，反应温度60℃，pH 4.5，反应时间60h，底物浓度70%（w/v）；糖化酶水解G2-β-CD的最佳条件为反应时间8h，加酶量40U/mL。酶处理后的混合产物可以经Sephadex G-25凝胶柱一步纯化及利用HPLC、LC-MS/MS以及NMR色谱对纯化产物鉴定结果证实该方法可以得到G1-β-CD纯品。
生物科学研究方法、生物科学研究技术药学生物化学
一锅法普鲁兰酶糖化酶葡萄糖基-β-环糊精麦芽糖基-β-环糊精凝胶柱层析
徐学明,柏玉香,金征宇,夏柳溪,王金鹏,吴云.葡萄糖基-β-环糊精的“一锅法”合成及其分离纯化[EB/OL].(2017-07-17)[2025-10-27].http://www.paper.edu.cn/releasepaper/content/201707-62.点此复制
Glucosyl-β-cyclodextrin (G1-β-CD) possesses better solubility and biocompatibility than parent β-cyclodextrin (β-CD) and has wide application prospects as it can be used in food, pharmaceutical, agriculture and chemical industry. But, because of the difficulties on the preparation and separation of G1-β-CD, few researchers and manufacturers have investigated and produced it currently. In this paper, the synthesis and separation of G1-β-CD were studied. Maltosyl-β-cyclodextrin (G2-β-CD) was obtained by grafting maltose (G2) onto β-CD by the reverse synthesis capability of pullulanase. Then G1-β-CD was obtained through the hydrolysis by glucoamylase in the same system without changing any reaction conditions. Afterwards, it was further purified by gel column. The results showed that the optimal conditions for the reverse synthesis of pullulanase were MG2: Mβ-CD of 8: 1, enzyme concentration of 360 U/g β-CD, reaction temperature of 60℃, pH of 4.5, reaction time of 60 h . The optimal conditions for G2-β-CD hydrolysis by glucoamylase were reaction time of 8h, enzyme concentration of 40 U/mL. Finally, the G1-β-CD samples were separated by Sephadex G-25 gel column and identified by HPLC, LC-MS / MS and NMR chromatography. The results confirmed that the yielded product was pure G1-β-CD.
One-pot SynthesispullulanaseGlucoamylaseGlucosyl-β-cyclodextrinMaltosyl-β-cyclodextringel column chromatography
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