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- Title
Generation of Highly Specific Proteolytic Biocatalysts by Screening Technologies.
- Authors
Bobik, T. V.; Kostin, N. N.; Knorre, V. D.; Gabibov, A. G.; Smirnov, I. V.
- Abstract
We propose a yeast display-based system for screening of proteolytic enzyme libraries that utilizes substrate protein adsorbed on the yeast cell surface and containing a desired cleavage sequence. Specific cleavage of the substrate protein releases its biotin-binding center. The cells carrying the target proteinase can be selected by cytofluorometry due to interaction with biotinylated fluorescent protein. Using human enterokinase light chain as the model proteinase we showed that the proposed screening system highly effectively selects the proteolytic enzymes with preset specificity.
- Subjects
PROTEOLYTIC enzymes; CELL membranes; PROTEIN engineering; GENE expression; MUTAGENESIS
- Publication
Bulletin of Experimental Biology & Medicine, 2018, Vol 165, Issue 3, p399
- ISSN
0007-4888
- Publication type
Article
- DOI
10.1007/s10517-018-4179-6