4LR9

Phosphopentomutase S154A variant soaked with 2,3-dideoxyribose 5-phosphate

4LR9 の概要

• エントリーDOI: 10.2210/pdb4lr9/pdb
• 関連するPDBエントリー: 3M8Z 3TWZ 3TX0 3UN3 4LR7 4LR8 4LRA 4LRB 4LRC 4LRD 4LRE 4LRF
• 分子名称: Phosphopentomutase, MANGANESE (II) ION, GLYCEROL, ... (5 entities in total)
• 機能のキーワード: alkaline phosphatase family, isomerase
• 由来する生物種: Bacillus cereus
• タンパク質・核酸の鎖数: 3
• 化学式量合計: 140324.80

構造登録者

Birmingham, W.A.,Starbird, C.A.,Panosian, T.D.,Nannemann, D.P.,Iverson, T.M.,Bachmann, B.O. (登録日: 2013-07-19, 公開日: 2013-07-31, 最終更新日: 2024-10-16)

主引用文献

Birmingham, W.R.,Starbird, C.A.,Panosian, T.D.,Nannemann, D.P.,Iverson, T.M.,Bachmann, B.O.
Bioretrosynthetic construction of a didanosine biosynthetic pathway.
Nat.Chem.Biol., 10:392-399, 2014

PubMed 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.

PubMed: 24657930
DOI: 10.1038/nchembio.1494

実験手法

X-RAY DIFFRACTION (2.1 Å)