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An Immobilized Biotin Ligase: Surface Display of Escherichia coli BirA on Saccharomyces cerevisiae
Author(s) -
Parthasarathy Ranganath,
Bajaj Jitin,
Boder Eric T.
Publication year - 2005
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp050279t
Subject(s) - dna ligase , biotinylation , biotin , saccharomyces cerevisiae , yeast , escherichia coli , biochemistry , enzyme , saccharomyces , biology , chemistry , gene
The Escherichia coli biotin ligase enzyme BirA has been extensively used in recent years to generate site‐specifically biotinylated proteins via a biotin acceptor peptide tag. In the present study, BirA was displayed for the first time on the yeast Saccharomyces cerevisiae using the Aga1p‐Aga2p platform and assayed using a peptide‐tagged protein as the substrate. The enzyme is fully functional and resembles the soluble form in many of its properties, but the yeast‐displayed enzyme demonstrates stability and reusability on the time scale of weeks. Thus, the yeast‐displayed BirA system represents a facile and highly economical alternative for producing site‐specifically biotinylated proteins.