4lrf
From Proteopedia
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== Structural highlights == | == Structural highlights == | ||
[[4lrf]] is a 3 chain structure with sequence from [http://en.wikipedia.org/wiki/Atcc_14579 Atcc 14579]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4LRF OCA]. <br> | [[4lrf]] is a 3 chain structure with sequence from [http://en.wikipedia.org/wiki/Atcc_14579 Atcc 14579]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=4LRF OCA]. <br> | ||
| - | <b>Related:</b> [[3un3|3un3]], [[3tx0|3tx0]], [[3twz|3twz]], [[3m8z|3m8z]], [[4lr7|4lr7]], [[4lr8|4lr8]], [[4lr9|4lr9]], [[4lra|4lra]], [[4lrb|4lrb]], [[4lrc|4lrc]], [[4lrd|4lrd]], [[4lre|4lre]]<br> | + | <b>[[Ligand|Ligands:]]</b> <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene>, <scene name='pdbligand=HSX:5-O-PHOSPHONO-ALPHA-D-RIBOFURANOSE'>HSX</scene>, <scene name='pdbligand=MN:MANGANESE+(II)+ION'>MN</scene><br> |
| + | <b>[[Non-Standard_Residue|NonStd Res:]]</b> <scene name='pdbligand=TPO:PHOSPHOTHREONINE'>TPO</scene><br> | ||
| + | <b>[[Related_structure|Related:]]</b> [[3un3|3un3]], [[3tx0|3tx0]], [[3twz|3twz]], [[3m8z|3m8z]], [[4lr7|4lr7]], [[4lr8|4lr8]], [[4lr9|4lr9]], [[4lra|4lra]], [[4lrb|4lrb]], [[4lrc|4lrc]], [[4lrd|4lrd]], [[4lre|4lre]]<br> | ||
<b>Activity:</b> <span class='plainlinks'>[http://en.wikipedia.org/wiki/Glucokinase Glucokinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.2 2.7.1.2] </span><br> | <b>Activity:</b> <span class='plainlinks'>[http://en.wikipedia.org/wiki/Glucokinase Glucokinase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.7.1.2 2.7.1.2] </span><br> | ||
| + | <b>Resources:</b> <span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=4lrf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=4lrf OCA], [http://www.rcsb.org/pdb/explore.do?structureId=4lrf RCSB], [http://www.ebi.ac.uk/pdbsum/4lrf PDBsum]</span><br> | ||
== Publication Abstract from PubMed == | == Publication Abstract from PubMed == | ||
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. | 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. | ||
Revision as of 10:13, 30 April 2014
Phosphopentomutase S154G variant soaked with ribose 5-phosphate
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