4lr7
Phosphopentomutase S154A variantPhosphopentomutase S154A variant
Structural highlights
FunctionDEOB_BACCR Phosphotransfer between the C1 and C5 carbon atoms of pentose (By similarity). Publication Abstract from PubMedConcatenation of engineered biocatalysts into multistep pathways markedly increases their utility, but the development of generalizable assembly methods remains a major 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 threefold 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. Bioretrosynthetic construction of a didanosine biosynthetic pathway.,Birmingham WR, Starbird CA, Panosian TD, Nannemann DP, Iverson TM, Bachmann BO Nat Chem Biol. 2014 Mar 23. doi: 10.1038/nchembio.1494. PMID:24657930[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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