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Functional display of target proteins on Bacillus subtilis spore and its application

机译：靶蛋白在枯草芽孢杆菌孢子上的功能展示及其应用

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@@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. 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.
机译：细菌表面展示在进化的酶，生物修复，全细胞生物转化和活疫苗生产工具的筛选工具领域中发现了重要的生物技术应用。对于多聚体形式的活性酶的功能性细菌表面展示，由于在细胞外分泌展示的靶蛋白后单体亚基的分子组装，通常是不可能的，因此开发了一种基于枯草芽孢杆菌孢子的新的表面展示系统。在检查的11种孢子外壳蛋白中，选择了cotE和cotG。使用该基序，仅以四聚体形式具有活性的β-半乳糖苷酶功能性地展示在枯草芽孢杆菌孢子的表面上。 β-半乳糖苷酶的表面定位通过纯化孢子的酶促测定，蛋白酶可及性测试和表达β-半乳糖苷酶的孢子的FACS分析进行验证。

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