阿糖胞苷
生物化学
大肠杆菌
嘌呤核苷磷酸化酶
化学
糖原磷酸化酶
酶
细胞
核苷酸回收
嘌呤
生物
基因
白血病
核苷酸
遗传学
作者
Li Ping,Jing Ruxian,Zhou Mengping,Jia Pei,Zhuoya Li,Guosheng Liu,Zhengyu Wang,Hailei Wang
摘要
Currently, whole-cell catalysts face challenges due to the complexity of reaction systems, although they have a cost advantage over pure enzymes. In this study, cytarabine was synthesized by purified purine phosphorylase 1 (PNP1) and uracil phosphorylase (UP), and the conversion of cytarabine from adenine arabinoside reached 72.3 ± 4.3%. However, the synthesis was unsuccessful by whole-cell catalysis due to interference from unnecessary proteins (UNPs) in cells. Thus, we carried out a large-scale gene editing involving 377 genes in the genome of Escherichia coli to reduce the negative effect of UNPs on substrate conversion and cytarabine production. Finally, the PNP1 and UP activities of the obtained mutant were increased significantly compared with the parental strain, and more importantly, the conversion rate of cytarabine by whole-cell catalysis reached 67.4 ± 2.5%. The lack of 148 proteins and downregulation of 783 proteins caused by gene editing were equivalent to partial purification of the enzymes within cells, and thus, we provided inspiration to solve the problem caused by UNP interference, which is ubiquitous in the field of whole-cell catalysis.
科研通智能强力驱动
Strongly Powered by AbleSci AI