State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China.
School of Food Science and Technology, Jiangnan University, Wuxi, China.
J Food Biochem. 2019 Aug;43(8):e12890. doi: 10.1111/jfbc.12890. Epub 2019 May 29.
Glucosyl-β-cyclodextrin (G -β-CD) has wide application prospects in the food, pharmaceutical, agriculture, and chemical industry as its better solubility and biocompatibility than parent β-cyclodextrin (β-CD). However, researches on G -β-CD were few due to difficulties in its preparation and separation. In the present study, a one-pot method was as follows: first, maltosyl-β-cyclodextrin (G -β-CD) was prepared through the reverse synthesis capability of pullulanase. Then, G -β-CD was obtained through hydrolysis by glucoamylase in the same reaction system and further purified by gel filtration chromatography. The results revealed that the optimal conditions for the reverse synthesis were as follows: n :n , 8:1; enzyme concentration, 60 U/ml; reaction temperature, 60°C; pH, 4.5; reaction time, 60 hr. The optimal conditions for G -β-CD hydrolysis were as follows: reaction time, 8 hr; enzyme concentration, 40 U/ml. Finally, the pure G -β-CD samples were obtained by Sephadex G-25 gel column separation and identified by HPLC, LC-MS/MS, and NMR chromatography. PRACTICAL APPLICATIONS: Commercially available modified CDs are almost synthesized using the chemical method, such as methyl-, sulfonylbutyl-, and hydroxypropyl-CD. However, the chemical synthesis of modified CDs has some security issue, such as organic solvent residue, which limits its application in foods. Based on the relevance references, G1-β-CD not only has the inclusion properties of the parent β-CD but also improves solubility and reduces the hemolysis effect and nephrotoxicity. However, the synthesis and separation of G1-β-CD is an obstacle to its expansion in production and application. In addition, there are few studies on G1-β-CD and few companies selling the product on the market. In this manuscript, the G1-β-CD was successfully synthesized by one-pot method and the optimal preparation and separation conditions of G1-β-CD were investigated.
葡糖基-β-环糊精（G-β-CD）比母体β-环糊精（β-CD）具有更好的溶解度和生物相容性，因此在食品、制药、农业和化学工业中具有广泛的应用前景。然而，由于其制备和分离困难，对 G-β-CD 的研究较少。在本研究中，采用一锅法：首先，通过普鲁兰酶的反合成能力制备麦芽葡糖基-β-环糊精（G-β-CD）；然后，在同一反应体系中通过糖化酶水解得到 G-β-CD，并进一步通过凝胶过滤色谱法纯化。结果表明，反合成的最佳条件为：n:n=8:1；酶浓度，60 U/ml；反应温度，60°C；pH 值，4.5；反应时间，60 小时。G-β-CD 水解的最佳条件为：反应时间 8 小时；酶浓度 40 U/ml。最后，通过 Sephadex G-25 凝胶柱分离得到纯 G-β-CD 样品，并通过 HPLC、LC-MS/MS 和 NMR 色谱进行鉴定。实际应用：市售的改性 CDs 几乎都是采用化学方法合成的，如甲基-β-CD、磺丁基-β-CD 和羟丙基-β-CD。然而，改性 CDs 的化学合成存在一些安全问题，如有机溶剂残留，限制了其在食品中的应用。基于相关参考文献，G1-β-CD 不仅具有母体β-CD 的包合性能，而且提高了溶解度，降低了溶血作用和肾毒性。然而，G1-β-CD 的合成和分离是其在生产和应用中扩大的障碍。此外，关于 G1-β-CD 的研究较少，市场上销售该产品的公司也较少。在本手稿中，通过一锅法成功合成了 G1-β-CD，并研究了 G1-β-CD 的最佳制备和分离条件。
