In vitro selection for catalytic turnover from a library of β-lactamase mutants and penicillin-binding proteins
✍ Scribed by Bérangère Avalle; Sophie Vanwetswinkel; Jacques Fastrez
- Book ID
- 104364854
- Publisher
- Elsevier Science
- Year
- 1997
- Tongue
- English
- Weight
- 346 KB
- Volume
- 7
- Category
- Article
- ISSN
- 0960-894X
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✦ Synopsis
A library of mutants of the RTEM 13-1actamase displayed on phage was created; it contained penicillin binding proteins (PBPs) as well as a small fraction of active B-lactamases. The library was submitted to a selection process to extract the B-lactamases i.e. the enzymes that turnover efficiently. This was achieved by a two steps procedure. In the first step, the 13-1actamases were labelled by reaction with a biotinylated suicide inhibitor while the PBPs were blocked by incubation in the presence of benzylpenicillin. In the second step, the labelled active phage-enzymes were separated by affinity chromatography on streptavidin coated beads.