Clone and Expression of High Yield Recombinant Trehalose Synthase in Bacillus subtilis

Trehalose synthase is one kind of intermolecular transglucosylation enzyme, which catalyzes the conversion of maltose to trehalose. In this study the trehalose synthase gene was amplified from Pseudomonas putida P06 genomic DNA, ligated with pMA5 vector, cloned into Bacillus subtilis WB800 and had good expression with the molecular weight of 77 KD. The recombinant trehalose synthase expression conditions were performed and enzyme reaction key parameters were investigated. The results showed the enzyme had optimal activity when it reacted for 2 h at 35 degrees C with pH value of 7.5 and the substrate concentration was 30 %. Trehalose content of samples was detected by HPLC and the enzyme activity reached to 318.12 U/ml in crude enzyme solution. This study is the first report about the expression of trehalose synthase in Bacillus subtilis, which lays the basis for trehalose large scale industrial production.