Functional Display of Active β-Galactosidase on Bacillus subtilis Spores Using Crust Proteins as Carriers

다국어 초록 (Multilingual Abstract)

Two model proteins of an enhanced green fluorescent protein (EGFP) and a tetrameric β- Galactosidase (β-Gal) from Bacillus stearothermophilus IAM11001 were fused to the C-terminus of the crust proteins Y (CotY) and Z (CotZ) from B. subtilis 168. Surface-expression of the resulting fusion proteins CotY-EGFP, CotZ-EGFP, CotY-β-Gal, and CotZ-β-Gal were then evaluated. Fluorescence intensity analysis of the transformed strains indicated that the CotY-EGFP and CotZ-EGFP fusion proteins were successfully displayed on spores. β-Gal was anchored on the surface of B. subtilis spores, confirmed based on western blotting and immunofluorescence microscopy. Results of a β-Gal assay showed that when fused to CotY and CotZ, the obtained enzyme activities of spore-displayed β-Gals were 0.12 and 0.25 U/mg of spores (dry weight), respectively. CotY and CotZ were, thus, shown to be suitable candidate carriers for display of multimeric enzymes on the spore surface.

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