earticle

영어

Bacterial surface display finds its important biotechnological application in the fields of screening tools of evolved enzyme, bioremediation, whole cell bioconversion and tool for live vaccine production [1]. 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 Bacillus subtilis spore was developed. Among 11 spore coat proteins examined, cotE and cotG were selected. Using this motif, beta-galactosidase, which is active only in tetrameric form, was functionally displayed on the surface of Bacillus subtilis spore. The surface localization of beta-galactosidase was verified by enzymatic assay of purified spore, protease accessibility test and FACS analysis of spore expressing beta-galactosidase. While Bacillus 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. Other enzymes such as streptavidin, lipase (lipB) of Bacillus subtilis and GFPUV were also successfully displayed using cotE and cotG motifs, and its possible application in various biotechnological fields will be discussed [2, 3]