Surface Display of Tres Using Cot C as a Molecular Vector on Bacillus subtilis Spores
ZHAO Yi-jin;WANG Teng-fei;WANG Jun-qing;WANG Rui-ming;QILU University of Technology;Department of Biology Engineering;
Trehalose is a ubiquitous non-reducing disaccharide in nature,and is an excellent natural drying agent and preservative. Trehalose synthase capable of catalyzing the maltose directly into trehalose,and is the preferred production of trehalose. To obtain trehalose synthase having good catalytic surface,which is displayed in a highly efficient and stable surface of Bacillus subtilis,at the same experiments were selected enhanced green fluorescent protein( EGFP) and trehalose synthase( Tres) as a model protein,to come from Bacillus subtilis spore coat protein Cot C as Bacillus subtilis anchored proteins displayed on the surface. Flow cytometry analysis of the situation in the spore surface display EGFP,the results showed that the capsid protein of Bacillus Cot C can EGFP fixed spore surface. Then replace fluorescent protein gene egfp and trehalose synthase gene tres. The recombinant strains was hang up using p H 7. 5 buffer suspension and the concentration of substrate for 30% of the maltose in 50℃ water bath roling 2h. Reaction products were analyzed by HPLC and the enzymatic activity can be detected,the enzyme activity of trehalose by calculating reached 252 U / ml. This suggests that Cot C is associated with the outer part of the coat. Cot C can therefore be used as a molecular vehicle for spore surface display of exogenous proteins.