Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase
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 a...
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| 格式: | Journal Article |
| 语言: | English |
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2022
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| 主题: | |
| 在线阅读: | https://hdl.handle.net/10356/163555 |
| _version_ | 1826124334556512256 |
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| author | Li, Ping Jing, Ruxian Zhou, Mengping Jia, Pei Li, Zhuoya Liu, Guosheng Wang, Zhenyu Wang, Hailei |
| author2 | Nanyang Environment and Water Research Institute |
| author_facet | Nanyang Environment and Water Research Institute Li, Ping Jing, Ruxian Zhou, Mengping Jia, Pei Li, Zhuoya Liu, Guosheng Wang, Zhenyu Wang, Hailei |
| author_sort | Li, Ping |
| collection | NTU |
| description | 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. |
| first_indexed | 2024-10-01T06:18:39Z |
| format | Journal Article |
| id | ntu-10356/163555 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T06:18:39Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | ntu-10356/1635552022-12-09T01:46:27Z Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase Li, Ping Jing, Ruxian Zhou, Mengping Jia, Pei Li, Zhuoya Liu, Guosheng Wang, Zhenyu Wang, Hailei Nanyang Environment and Water Research Institute Advanced Environmental Biotechnology Centre (AEBC) Science::Biological sciences Cytarabine Escherichia Coli 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. This study was supported by the Excellent Scientific and Technological Innovation Team of Henan Normal University, China (5101049170501) and the National Science Foundations of China (U160411067). 2022-12-09T01:46:26Z 2022-12-09T01:46:26Z 2022 Journal Article 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. https://dx.doi.org/10.1002/bit.28098 0006-3592 https://hdl.handle.net/10356/163555 10.1002/bit.28098 35383880 2-s2.0-85128072813 7 119 1768 1780 en Biotechnology and Bioengineering © 2022 Wiley Periodicals LLC. All rights reserved. |
| spellingShingle | Science::Biological sciences Cytarabine Escherichia Coli Li, Ping Jing, Ruxian Zhou, Mengping Jia, Pei Li, Zhuoya Liu, Guosheng Wang, Zhenyu Wang, Hailei Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase |
| title | Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase |
| title_full | Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase |
| title_fullStr | Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase |
| title_full_unstemmed | Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase |
| title_short | Whole-cell biosynthesis of cytarabine by an unnecessary protein-reduced Escherichia coli that coexpresses purine and uracil phosphorylase |
| title_sort | whole cell biosynthesis of cytarabine by an unnecessary protein reduced escherichia coli that coexpresses purine and uracil phosphorylase |
| topic | Science::Biological sciences Cytarabine Escherichia Coli |
| url | https://hdl.handle.net/10356/163555 |
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