Generation of Highly Specific Proteolytic Biocatalysts by Screening Technologies
- Authors: Bobik T.V.1, Kostin N.N.1, Knorre V.D.1, Gabibov A.G.1, Smirnov I.V.1
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Affiliations:
- Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Organic Biochemistry, Russian Academy of Sciences
- Issue: Vol 165, No 3 (2018)
- Pages: 399-402
- Section: Article
- URL: https://journals.rcsi.science/0007-4888/article/view/240354
- DOI: https://doi.org/10.1007/s10517-018-4179-6
- ID: 240354
Cite item
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.
About the authors
T. V. Bobik
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Organic Biochemistry, Russian Academy of Sciences
Author for correspondence.
Email: bobik_tanya@mail.ru
Russian Federation, Moscow
N. N. Kostin
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Organic Biochemistry, Russian Academy of Sciences
Email: bobik_tanya@mail.ru
Russian Federation, Moscow
V. D. Knorre
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Organic Biochemistry, Russian Academy of Sciences
Email: bobik_tanya@mail.ru
Russian Federation, Moscow
A. G. Gabibov
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Organic Biochemistry, Russian Academy of Sciences
Email: bobik_tanya@mail.ru
Russian Federation, Moscow
I. V. Smirnov
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Organic Biochemistry, Russian Academy of Sciences
Email: bobik_tanya@mail.ru
Russian Federation, Moscow