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

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 aff...

Full description

Saved in:
Bibliographic Details
Published in:Angewandte Chemie 2016-07, Vol.128 (30), p.8727-8731
Main Authors: Rosen, Christian B, Kwant, Richard L, MacDonald, James I, Rao, Meera, Francis, Matthew B
Format: Article
Language:eng ; ger
Subjects:
Affinity
Bile
Binding
Chemistry
Conjugates
Constraining
Evolution
Polymers
Recycling
Strategy
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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 sortaseA (SrtA7M) was N-terminally labeled with lithocholic acid (LA)--an inexpensive bile acid that exhibits strong binding to [beta]-cyclodextrin ([beta]CD). Capture and recycling of the LA-Pro-SrtA7M conjugate was achieved using [beta]CD-modified sepharose resin. The LA-Pro-SrtA7M conjugate retained full enzymatic activity, even after multiple rounds of recycling.
ISSN:0044-8249
1521-3757
DOI:10.1002/ange.201602353