A Fructan Sucrase Secreted Extracellular and Purified in One-Step by Gram-Positive Enhancer Matrix Particles

Fructan sucrase is a kind of biological enzyme that catalyzes the synthesis fructan, and fructan polysaccharide product with important industrial application value. In this study, gene Bacillus subtilis was cloned to plasmid PET-28A-ACMA-Z, three clones were obtained after transformation Escherichia coli BL21, namely BS-FF, BSO, BS. The BS-FF BSO secreted recombinant enzymes outside cells, while clone BS expressed them inside cells. induction experiment results showed optimum IPTG concentration in medium 0.5 mM 1.0 for respectively, incubation conditions at 28 °C 8 h. purified one step using material called GEM particles. indicated 95.25% by could be into extracellular area, even 98.78% BSO. With above purification system, receiving rate 97.70% 84.99%, respectively. As bioactivity sucrase, temperature pH 50 5.6, Km Vmax it 33.96 g/L 0.63 g/(L·min), engineered strains high secretion constructed, one-step method established. might provide novel selection enzymatic production on large scale.