Nowadays enzymatic synthesis of nucleic acid derivatives is gaining momentum over traditional chemical synthetic processes. Biotransformations catalyzed by whole cells or enzymes offer an ecofriendly and efficient alternative to the traditional multistep chemical methods, avoiding the use of chemical reagents and organic solvents that are expensive and environmentally harmful. Herein we report for the first time the covalent immobilization a uracil phosphoribosyltransferase (UPRT). In this sense, UPRT from Thermus thermophilus HB8 was immobilized onto glutaraldehyde-activated MagReSyn®Amine magnetic iron oxide porous microparticles (MTtUPRT). According to the catalyst load experiments, MTtUPRT3 was selected as optimal biocatalyst for further studies. MTtUPRT3 was active and stable in a broad range of temperature (70-100 °C) and in the pH interval 6-8, displaying maximum activity at 100 °C and pH 7 (activity 968 IU/gsupport, retained activity 100%). In addition, MTtUPRT3 could be reused up to 8 times in the synthesis of uridine-5'-monophosphate (UMP). Finally, MTtUPRT3 was successfully applied in the sustainable synthesis of different 5-modified uridine-5'-monophosphates at short times. Taking into account these results, MTtUPRT3 would emerge as a valuable biocatalyst for the synthesis of nucleoside monophosphates through an efficient and environmentally friendly methodology.

中文翻译：

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固定化的嗜热栖热菌尿嘧啶磷酸核糖基转移酶可持续合成尿苷5'-单磷酸酯类似物。

如今，核酸衍生物的酶促合成已超过了传统的化学合成工艺。全细胞或酶催化的生物转化为传统的多步化学方法提供了一种生态友好且高效的替代方法，从而避免了使用昂贵且对环境有害的化学试剂和有机溶剂。在此，我们首次报道了尿嘧啶磷酸核糖基转移酶（UPRT）的共价固定。在这个意义上，将嗜热栖热菌HB8的UPRT固定在戊二醛活化的MagReSyn®Amine磁性氧化铁多孔微粒（MTtUPRT）上。根据催化剂负载实验，选择MTtUPRT3作为进一步研究的最佳生物催化剂。MTtUPRT3在很宽的温度范围（70-100°C）和pH区间6-8中均具有活性和稳定性，在100°C和pH 7时显示最大活性（活性968 IU / g支持，保留活性100％）。此外，MTtUPRT3在尿苷5'-单磷酸酯（UMP）的合成中最多可重复使用8次。最终，MTtUPRT3成功地用于短时间内可持续合成不同的5-修饰的尿苷5'-单磷酸酯。考虑到这些结果，MTtUPRT3将成为通过有效且环保的方法合成核苷一磷酸的有价值的生物催化剂。最终，MTtUPRT3成功地用于短时间内可持续合成不同的5-修饰的尿苷5'-单磷酸酯。考虑到这些结果，MTtUPRT3将成为通过有效且环保的方法合成核苷一磷酸的有价值的生物催化剂。最终，MTtUPRT3成功地用于短时间内可持续合成不同的5-修饰的尿苷5'-单磷酸酯。考虑到这些结果，MTtUPRT3将成为通过有效且环保的方法合成核苷一磷酸的有价值的生物催化剂。