以量大易得的莱鲍迪苷A(rebaudioside A)为底物通过酶法制备莱鲍迪苷D(rebaudioside D)是目前获得莱鲍迪苷D的主要途径,但是从复杂的酶催化体系中富集并分离莱鲍迪苷D较为困难。为实现酶催化体系中莱鲍迪苷D的有效富集与分离,该研究选取了3种对甜菊糖核心骨架贝壳杉烯有吸附能力的苯乙烯-二乙烯苯骨架的大孔树脂。考察了3种树脂富集莱鲍迪苷与分离莱鲍迪苷D的能力,选出了能够有效吸附莱鲍迪苷D并对莱鲍迪苷D具有良好分离能力的LX-T28树脂。最后,该研究以LX-T28树脂为填料采用连续柱色谱法实现了酶催化体系中莱鲍迪苷D的富集与分离,高收率低成本地获得了纯度＞95%的莱鲍迪苷D,为后续酶法生产莱鲍迪苷D实现工业化解决了后处理难题。
The conversion of Rebaudioside D from Rebaudioside A, which is an abundant and readily available natural source, can be realized by enzyme catalytic conversion.However, the enrichment and separation of Rebaudioside D from the complex enzymatic reaction solution remains still a challenge.To solve this problem, three styrene-divinylbenzene macroporous resins known for their adsorption capacity for the core framework kaurene of stevioside were selected.The ability of these three materials to enrich rebaudioside D and separate Rebaudioside D was evaluated, and the LX-T28 resin was finally chosen for its effective absorption and separation capability among them.Ultimately, a method which employed tandem column chromatography for the separation of Rebaudioside D from the enzymatic reaction system was demonstrated in this study.A Rebaudioside D production with a purity exceeding 95% was obtained at a high yield and low cost.In this research the post-reaction issue of Rebaudioside D production was resolved and perhaps this paves the way for the commercial supply of Rebaudioside D.
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