Bioretrosynthetic construction of a didanosine biosynthetic pathway
| Autor: | William R Birmingham, T.D. Panosian, Brian O. Bachmann, D.P. Nannemann, Tina M. Iverson, Chrystal A Starbird |
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| Rok vydání: | 2014 |
| Předmět: |
Models
Molecular chemistry.chemical_classification Purine nucleoside phosphorylase Cell Biology Isomerase Biology Phosphopentomutase Directed evolution Article Enzymes Didanosine Enzyme Biochemistry chemistry Biocatalysis medicine Directed Molecular Evolution Ribokinase Molecular Biology Nucleoside medicine.drug |
| Zdroj: | Nature chemical biology Birmingham, W R, Starbird, C A, Panosian, T D, Nannemann, D P, Iverson, T M & Bachmann, B O 2014, ' Bioretrosynthetic construction of a didanosine biosynthetic pathway ', Nature chemical biology, vol. 10, no. 5, pp. 392-399 . https://doi.org/10.1038/nchembio.1494 |
| ISSN: | 1552-4469 1552-4450 |
| DOI: | 10.1038/nchembio.1494 |
| Popis: | Concatenation of engineered biocatalysts into multistep pathways dramatically increases their utility, but development of generalizable assembly methods remains a significant challenge. Herein we evaluate ‘bioretrosynthesis’, which is an application of the retrograde evolution hypothesis, for biosynthetic pathway construction. To test bioretrosynthesis, we engineered a pathway for synthesis of the antiretroviral nucleoside analog didanosine (2,3-dideoxyinosine). Applying both directed evolution and structure-based approaches, we began pathway construction with a retro-extension from an engineered purine nucleoside phosphorylase and evolved 1,5-phosphopentomutase to accept the substrate 2,3-dideoxyribose 5-phosphate with a 700-fold change in substrate selectivity and 3-fold increased turnover in cell lysate. A subsequent retrograde pathway extension, via ribokinase engineering, resulted in a didanosine pathway with a 9,500-fold change in nucleoside production selectivity and 50-fold increase in didanosine production. Unexpectedly, the result of this bioretrosynthetic step was not a retro-extension from phosphopentomutase, but rather the discovery of a fortuitous pathway-shortening bypass via the engineered ribokinase. |
| Databáze: | OpenAIRE |
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