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Display of large enzyme complexes on the cell surface of Bacillus subtilis
Author(s) -
Tjia Wai Mui,
Lindquist Erica,
Wu Sauching,
Wong Suilam
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.lb24-d
Subject(s) - streptavidin , linker , biotinylation , bacillus subtilis , chemistry , biochemistry , cell wall , hydrolase , enzyme , cell , protein subunit , fusion protein , biotin , biophysics , biology , recombinant dna , bacteria , computer science , gene , genetics , operating system
The ability to display multi‐subunit enzyme complexes on the bacterial cell surface offers opportunities to degrade complex polymers such as plant cellulose in an efficient and cooperative manner. To develop an engineered nanomachinery for surface display of enzymes, a fusion protein consists of streptavidin followed by a flexible linker and a cell wall binding domain derived from Bacillus subtilis cell wall hydrolase, LytE, is designed. Since each streptavidin subunit can self‐assemble into the tetrameric state and capture an enzyme (β‐lactamase in this study) carrying a streptavidin binding tag (SBP), a 140‐kDa complex is expected to form. Also, the tetrameric streptavidin fusion can potentially enhance the cell wall binding avidity. To examine the display efficiency, linkers with different lengths (51 and 151 amino acids) in the streptavidin fusions are designed. SDS‐PAGE results showed that the streptavidin fusions could be displayed as tetramers on the cell surface. Enzyme assay and Western blot showed that SBP‐tagged β‐lactamase could be captured to the cell surface. Interestingly, cells displaying streptavidin fusions with a longer linker showed 44% higher β‐lactamase activity. Longer linker is suggested to offer better surface accessibility of the displayed streptavidin fusions. This system can potentially be applied to display one or multiple species of SBP‐tagged proteins on the cell surface.