| 초록 |
For the functional bacterial surface display of active enzyme of multimeric form, which is generally impossible due to molecular assembly of the monomer subunit subsequent to the secretion of displayed target protein outside the cell, a new surface display system based on B. subtilis spore was developed. Among 11 spore coat proteins examined, cotE and cotG were selected. Using this motif, b-galactosidase, which is active in dimeric or tetrameric form, was functionally displayed on the surface of B. subtilis spore. The surface localization of b-galactosidase was verified by PAGE analysis of spore-extracted fraction, enzymatic assay of purified spore, protease accessibility test and FACS analysis of spore expressing b-galactosidase. While B. subtilis spore wall integrity, examined by lysozyme and heat treatments, was slightly affected by the incorporation of CotE-LacZ fusion protein, it was not affected by the incorporation of CotG-lacZ fusion. |
