Abstract
Fatty acid binding proteins are a class of small 15 kDa proteins with a simple architecture that forms a large solvent sequestered cavity. In previous work, we demonstrated that reductive amination reactions could be performed in this cavity by covalent attachment of a pyridoxamine cofactor to the protein. Here, we report the results of experiments in which the position of pyridoxamine attachment has been varied by site directed mutagenesis. The conjugate IFABP-PX60 reacts at least 9.4-fold more rapidly than our original conjugate ALBP-PX, while IFABP-PX72 inverts the enantioselectivity of reactions (compared to ALBP-PX) and IFABP-PX104 displays very selective substrate specificity. These results indicate that site-directed mutagenesis can be used to tune the rate, enantioselectivity, and substrate specificity of semisynthetic transaminases based on fatty acid binding proteins.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2055-2060 |
| Number of pages | 6 |
| Journal | Bioorganic and Medicinal Chemistry Letters |
| Volume | 7 |
| Issue number | 15 |
| DOIs | |
| State | Published - Aug 5 1997 |
Bibliographical note
Funding Information:Acknowledgements. We thankD r. C. Friedenfo r providingth ep lasmides ncodingth em utanpt roteinus sed in thiss tudy.M odelingo f IFABP was performebdy M. Silberglitatn dS. Flaim. This work was supportebdy grants from the National ScienceF oundation( NSF-CHE-9506793)a nd the PetroleumR esearchF und (PRF#28140-G4).R . R. Davies was supportedb y a NationalI nstituteso f Health Training Grant (NIH 2T32GM08347-06).