Generation of Highly Specific Proteolytic Biocatalysts by Screening Technologies

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 t...

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Bibliographic Details
Published inBulletin of experimental biology and medicine Vol. 165; no. 3; pp. 399 - 402
Main Authors Bobik, T. V., Kostin, N. N., Knorre, V. D., Gabibov, A. G., Smirnov, I. V.
Format Journal Article
LanguageEnglish
Published New York Springer US 01.07.2018
Springer
Springer Nature B.V
Subjects
Aspartic proteases
Biocatalysts
Biomedical and Life Sciences
Biomedicine
Biotin
Calpain
Cell Biology
Cell surface
Cytofluorometry
Fluorescence
Health aspects
Health screening
Internal Medicine
Laboratory Medicine
Pathology
Protein binding
Proteinase
Proteins
Proteolysis
Proteolytic enzymes
Recombinant proteins
yeast display
screening of proteolytic enzymes for medical use
Pichia pastoris expression system
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Summary: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.
Bibliography:ObjectType-Article-1
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content type line 23
ISSN:0007-4888
1573-8221
DOI:10.1007/s10517-018-4179-6