4LR8: Phosphopentomutase S154a Variant Soaked With Ribose 5-phosphate

Citation:
Abstract
Concatenation 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.
PDB ID: 4LR8Download
MMDB ID: 112331
PDB Deposition Date: 2013/7/19
Updated in MMDB: 2013/08
Experimental Method:
x-ray diffraction
Resolution: 2  Å
Source Organism:
Similar Structures:
Biological Unit for 4LR8: monomeric; determined by author and by software (PISA)
Molecular Components in 4LR8
Label Count Molecule
Protein (1 molecule)
1
Phosphopentomutase(Gene symbol: BC4087)
Molecule annotation
Chemicals (7 molecules)
1
3
2
1
3
1
4
2
* Click molecule labels to explore molecular sequence information.

Citing MMDB
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