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Title: Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase
Authors: Li, Ping
Jing, Ruxian
Zhou, Mengping
Jia, Pei
Li, Zhuoya
Liu, Guosheng
Wang, Zhenyu
Wang, Hailei
Keywords: Science::Biological sciences
Issue Date: 2022
Source: Li, P., Jing, R., Zhou, M., Jia, P., Li, Z., Liu, G., Wang, Z. & Wang, H. (2022). Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase. Biotechnology and Bioengineering, 119(7), 1768-1780.
Journal: Biotechnology and Bioengineering
Abstract: 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.
ISSN: 0006-3592
DOI: 10.1002/bit.28098
Rights: © 2022 Wiley Periodicals LLC. All rights reserved.
Fulltext Permission: none
Fulltext Availability: No Fulltext
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