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Capture and Recycling of Sortase A through Site‐Specific Labeling with Lithocholic Acid
Author(s) -
Rosen Christian B.,
Kwant Richard L.,
MacDonald James I.,
Rao Meera,
Francis Matthew B.
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201602353
Subject(s) - sortase a , chemistry , conjugate , lithocholic acid , sortase , enzyme , biochemistry , limiting , combinatorial chemistry , biocatalysis , bile acid , catalysis , bacterial protein , mechanical engineering , mathematical analysis , ionic liquid , mathematics , engineering , gene
Enzyme‐mediated protein modification often requires large amounts of biocatalyst, adding significant costs to the process and limiting industrial applications. Herein, we demonstrate a scalable and straightforward strategy for the efficient capture and recycling of enzymes using a small‐molecule affinity tag. A proline variant of an evolved sortase A (SrtA 7M) was N‐terminally labeled with lithocholic acid (LA)—an inexpensive bile acid that exhibits strong binding to β‐cyclodextrin (βCD). Capture and recycling of the LA‐Pro‐SrtA 7M conjugate was achieved using βCD‐modified sepharose resin. The LA‐Pro‐SrtA 7M conjugate retained full enzymatic activity, even after multiple rounds of recycling.