Capture and Recycling of Sortase A through Site-Specific Labeling with Lithocholic Acid

Christian B Rosen; Richard L Kwant; James I MacDonald; Meera Rao; Matthew B Francis

doi:10.1002/anie.201602353

Capture and Recycling of Sortase A through Site-Specific Labeling with Lithocholic Acid

Angew Chem Int Ed Engl. 2016 Jul 18;55(30):8585-9. doi: 10.1002/anie.201602353. Epub 2016 May 30.

Authors

Christian B Rosen^{1

2}, Richard L Kwant^{1

2}, James I MacDonald^{1

2}, Meera Rao^{1

2}, Matthew B Francis^{3

4}

Affiliations

¹ Department of Chemistry, University of California, Berkeley, Berkeley, CA, 94720-1460, USA.
² Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
³ Department of Chemistry, University of California, Berkeley, Berkeley, CA, 94720-1460, USA. mbfrancis@berkeley.edu.
⁴ Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA. mbfrancis@berkeley.edu.

PMID: 27239057
DOI: 10.1002/anie.201602353

Abstract

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.

Keywords: affinity separation; bioconjugation; cyclodextrins; enzyme recycling; protein modification.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.