Henan Engineering Research Center of Bioconversion Technology of Functional Microbes, College of Life Science, Henan Normal University, Xinxiang 453007, China.
Biomolecules. 2024 Sep 3;14(9):1107. doi: 10.3390/biom14091107.
Arabinoside and derived nucleoside analogs, a family of nucleoside analogs, exhibit diverse typically biological activities and are widely used as antibacterial, antiviral, anti-inflammatory, antitumor, and other drugs in clinical and preclinical trials. Although with a long and rich history in the field of medicinal chemistry, the biosynthesis of arabinoside has only been sporadically designed and studied, and it remains a challenge. Here, we constructed an in vitro multi-enzymatic cascade for the biosynthesis of arabinosides. This artificial biosystem was systematically optimized, involving an exquisite pathway design, NADP regeneration, meticulous enzyme selection, optimization of the key enzyme dosage, and the concentration of inorganic phosphate. Under the optimized conditions, we achieved 0.37 mM of vidarabine from 5 mM of sucrose and 2 mM of adenine, representing 18.7% of the theoretical yield. Furthermore, this biosystem also has the capability to produce other arabinosides, such as spongouridine, arabinofuranosylguanine, hypoxanthine arabinofuranoside, fludarabine, and 2-methoxyadenine arabinofuranoside, from sucrose, and corresponding nucleobase by introducing different nucleoside phosphorylases. Overall, our biosynthesis approach provides a pathway for the biosynthesis of arabinose-derived nucleoside analogs, offering potential applications in the pharmaceutical industry.
阿拉伯糖苷和衍生的核苷类似物是一类核苷类似物，具有多样化的典型生物学活性，广泛应用于临床前和临床试验中的抗菌、抗病毒、抗炎、抗肿瘤等药物。尽管在药物化学领域有着悠久而丰富的历史，但阿拉伯糖苷的生物合成仅被零星设计和研究过，仍然是一个挑战。在这里，我们构建了一种用于阿拉伯糖苷生物合成的体外多酶级联反应。这个人工生物系统经过系统优化，涉及到精细的途径设计、NADP 再生、精心选择酶、关键酶剂量和无机磷酸盐浓度的优化。在优化条件下，我们从 5 mM 的蔗糖和 2 mM 的腺嘌呤中获得了 0.37 mM 的阿昔洛韦，理论产率为 18.7%。此外，该生物系统还能够通过引入不同的核苷磷酸化酶，从蔗糖和相应的核苷碱基中生产其他阿拉伯糖苷，如海绵尿苷、阿拉伯呋喃糖基鸟嘌呤、次黄嘌呤阿拉伯呋喃糖苷、氟达拉滨和 2-甲氧基腺嘌呤阿拉伯呋喃糖苷。总的来说，我们的生物合成方法为阿拉伯糖衍生的核苷类似物的生物合成提供了一种途径，为制药工业提供了潜在的应用。
