Department of Drug Sciences, Università degli Studi di Pavia, I-27100 Pavia, Italy.
Applied Biotechnology Group, Universidad Europea de Madrid, 28670 Villaviciosa de Odón, Spain; Grupo de Investigación en Ciencias Naturales y Exactas, GICNEX, Universidad de la Costa, CUC, 080003 Barranquilla, Atlántico, Colombia.
Bioresour Technol. 2020 Jul;307:123258. doi: 10.1016/j.biortech.2020.123258. Epub 2020 Mar 26.
In this work, a mono- and a bi-enzymatic analytical immobilized enzyme reactors (IMERs) were developed as prototypes for biosynthetic purposes and their performances in the in-flow synthesis of nucleoside analogues of pharmaceutical interest were evaluated. Two biocatalytic routes based on nucleoside 2'-deoxyribosyltransferase from Lactobacillus reuteri (LrNDT) and uridine phosphorylase from Clostridium perfrigens (CpUP)/purine nucleoside phosphorylase from Aeromonas hydrophila (AhPNP) were investigated in the synthesis of 2'-deoxy, 2',3'-dideoxy and arabinonucleoside derivatives. LrNDT-IMER catalyzed the synthesis of 5-fluoro-2'-deoxyuridine and 5-iodo-2'-deoxyuridine in 65-59% conversion yield, while CpUP/AhPNP-IMER provided the best results for the preparation of arabinosyladenine (60% conversion yield). Both IMERs proved to be promising alternatives to chemical routes for the synthesis of nucleoside analogues. The developed in-flow system represents a powerful tool for the fast production on analytical scale of nucleosides for preliminary biological tests.
在这项工作中，开发了单酶和双酶分析固定化酶反应器（IMER）作为生物合成的原型，并评估了它们在具有药物应用价值的核苷类似物的流动合成中的性能。研究了基于来自雷氏乳酸杆菌（LrNDT）的核苷 2'-脱氧核糖基转移酶和来自腐败梭菌（CpUP）/来自嗜水气单胞菌的嘌呤核苷磷酸化酶的两种生物催化途径，用于合成 2'-脱氧、2'，3'-二脱氧和阿拉伯糖核苷衍生物。LrNDT-IMER 催化 5-氟-2'-脱氧尿苷和 5-碘-2'-脱氧尿苷的合成，转化率为 65-59%，而 CpUP/AhPNP-IMER 为制备阿拉伯糖腺嘌呤提供了最佳结果（转化率为 60%）。两种 IMER 都被证明是化学途径合成核苷类似物的有前途的替代方法。所开发的流动系统代表了在分析规模上快速生产用于初步生物学测试的核苷的有力工具。
