title: Capture and Recycling of Sortase A through Site-Specific Labeling with Lithocholic Acid.
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.
authors.affiliation: Department of Chemistry, University of California, Berkeley, Berkeley, CA, 94720-1460, USA.|Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
authors.forename: Christian B
authors.initials: CB
authors.lastname: Rosen
authors.affiliation: Department of Chemistry, University of California, Berkeley, Berkeley, CA, 94720-1460, USA.|Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
authors.forename: Richard L
authors.initials: RL
authors.lastname: Kwant
authors.affiliation: Department of Chemistry, University of California, Berkeley, Berkeley, CA, 94720-1460, USA.|Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
authors.forename: James I
authors.initials: JI
authors.lastname: MacDonald
authors.affiliation: Department of Chemistry, University of California, Berkeley, Berkeley, CA, 94720-1460, USA.|Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA.
authors.forename: Meera
authors.initials: M
authors.lastname: Rao
authors.affiliation: Department of Chemistry, University of California, Berkeley, Berkeley, CA, 94720-1460, USA. mbfrancis@berkeley.edu.|Materials Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA. mbfrancis@berkeley.edu.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Francis
keywords: affinity separation; bioconjugation; cyclodextrins; enzyme recycling; protein modification