Enhancing and Reversing the Stereoselectivity of Escherichia coli Transketolase via Single-Point Mutations
Chiral auxiliary methodology and chiral assays have been developed to establish the enantiomeric purities of erythrulose and 1,3‐dihydroxypentan‐2‐one generated using wild‐type (WT) Escherichia coli transketolase (TK). L‐Erythrulose was formed in 95% ee and (3S)‐1,3‐dihydroxypentan‐2‐one in 58% ee....
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Published in | Advanced synthesis & catalysis Vol. 350; no. 16; pp. 2631 - 2638 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
Published |
Weinheim
WILEY-VCH Verlag
03.11.2008
WILEY‐VCH Verlag |
Subjects | |
Online Access | Get full text |
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Summary: | Chiral auxiliary methodology and chiral assays have been developed to establish the enantiomeric purities of erythrulose and 1,3‐dihydroxypentan‐2‐one generated using wild‐type (WT) Escherichia coli transketolase (TK). L‐Erythrulose was formed in 95% ee and (3S)‐1,3‐dihydroxypentan‐2‐one in 58% ee. Since the latter compound was formed in moderate ee, TK libraries were screened to identify higher performing mutants. A colorimetric screen and chiral assay were successfully applied to a 96‐well format, and new active TK mutants were identified, which gave 1,3‐dihydroxypentan‐2‐one in high stereoselectivities. Remarkably, active‐site single‐point mutants were identified that were able to both enhance and reverse the stereoselectivity of TK. |
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Bibliography: | ark:/67375/WNG-K778QBZ0-F EPSRC - No. GR/S62505/01 ArticleID:ADSC200800489 istex:904F3A0F8712F7ED2369C6FB7D203E4551578887 UK Engineering and Physical Sciences Research Council ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 |
ISSN: | 1615-4150 1615-4169 |
DOI: | 10.1002/adsc.200800489 |