State Key Laboratory of Chemical Resources Engineering, Beijing University of Chemical Technology, Beijing, 100029, People's Republic of China.
Appl Microbiol Biotechnol. 2013 Aug;97(15):6769-78. doi: 10.1007/s00253-012-4542-x. Epub 2012 Nov 20.
Biocatalyzed synthesis of nucleoside analogues was carried out using two thermostable nucleoside phosphorylases from the hyperthermophilic aerobic crenarchaeon Aeropyrum pernix K1. The synthesis of the 2,6-diaminopurine nucleoside and 5-methyluridine was used as a reaction model to test the process. Both the purine nucleoside phosphorylase (apPNP) and uridine phosphorylase (apUP) were functionally expressed in Escherichia coli. The recombinant enzymes were characterized after purification, and both enzymes showed high thermostability and broad substrate specificity. Both enzymes retained 100 % of their activity after 60 min at high temperature, and the optimum temperature for the enzymes was 90-100 °C. The nucleoside phosphorylases obtained from A. pernix are valuable industrial biocatalysts for high-temperature reactions that produce nucleoside drugs in high yields.
使用来自嗜热需氧古菌 Aeropyrum pernix K1 的两种耐热核苷磷酸化酶进行核苷类似物的生物催化合成。以 2,6-二氨基嘌呤核苷和 5-甲基尿嘧啶的合成为反应模型来测试该过程。嘌呤核苷磷酸化酶（apPNP）和尿苷磷酸化酶（apUP）均在大肠杆菌中实现了功能表达。在纯化后对重组酶进行了表征，两种酶均表现出高热稳定性和广泛的底物特异性。两种酶在高温下 60 分钟后仍保持 100%的活性，酶的最适温度为 90-100°C。从 A. pernix 获得的核苷磷酸化酶是用于高温反应的有价值的工业生物催化剂，可高产核苷药物。
