One-step enzymatic synthesis of nucleosides from low water-soluble purine bases in non-conventional media

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• 作者：Jesú ; s Ferná ; ndez-Lucas^a ; ^b ; Alba Fresco-Taboada^a ; Isabel de la Mata^a ; ¹ ; ^{isabel@bbm1.ucm.es} ; Miguel Arroyo^a ; ¹ ; ^{arroyo@bbm1.ucm.es}
• 关键词：2&prime ; -Deoxyribosyltransferase ; Lactobacillus reuteri ; Nucleoside synthesis ; Organic solvents ; Sepabeads
• 刊名：Bioresource Technology
• 出版年：2012
• 期刊代码：11_09608524
• 类别：ce
• 出版时间：July, 2012
• 卷：115
• 期：Complete
• 页码：63-69
• 文件大小：462 K

摘要

The effect of several water-miscible cosolvents on activity and stability of soluble and immobilized 2鈥?deoxyribosyltransferase from Lactobacillus reuteri on Sepabeads庐 has been studied in order to establish optimal conditions for enzymatic synthesis of nucleosides using purine bases with low solubility in aqueous buffer. As a rule of thumb, there was a general reduction of soluble enzyme activity when cosolvent content was gradually increased in reaction medium. In contrast, immobilized enzyme activity was enhanced 1.2-1.4-fold at 20%of methanol, ethanol, 2-propanol, diethylene glycol, and acetone; and at 10%and 30%acetonitrile. Likewise, highest increased activity (1.8-fold) was also obtained in presence of 20%acetonitrile. Immobilized enzyme was successfully used in the synthesis of 2鈥?deoxyxanthosine and 2鈥?deoxyguanosine using 2鈥?deoxyuridine as sugar donor and the corresponding poor water-soluble base in the presence of 30%of methanol, ethanol, 2-propanol, ethylene glycol, acetonitrile, and DMSO, giving high nucleoside yields at 4 h.