Characterization of levansucrase produced by novel Bacillus siamensis and optimization of culture condition for levan biosynthesis

Abstract

Levan has attracted interest due to the potential health benefits associated with its prebiotic, biological, and functional properties. However, the production of levan is expensive due to its high resource requirements. With the growing demand for levan, it is vital to determine suitable cultivation condition for its production and reduce costs accordingly. The present study characterized the enzyme levansucrase produced by a novel strain of Bacillus siamensis and optimized the conditions for the biosynthesis of levansucrase and levan. The crude levansucrase enzyme production by B. siamensis was induced at a specific temperature in a medium containing different concentrations of sucrose, fructose, and glucose to evaluate transfructosylation and hydrolysis activities. Crude levansucrase significantly increased transfructosylation relative to hydrolysis activity at 37 °C in a medium containing 20% (w/v) sucrose. Both transfructosylation and hydrolysis activities were inhibited in glucose and fructose containing medium. Purification and characterization of the levansucrase were performed by precipitating the enzyme with ammonium sulfate solution, purified anion-exchange chromatography, and analyzed by Sodium Dodecyl Sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The results showed the molecular weight of the enzyme to be approximately 30 kDa with specific activity at 15.95 U/mg, corresponding to a protein purification efficiency of 11.47 and a yield of 78.75%. The optimal culture condition for the purified-levansucrase activity for levan biosynthesis was obtained at 37 °C after 48 h, at pH 6.0 in 50 mM phosphate buffer and 20% (w/v) sucrose. The study demonstrated the optimized condition for levan biosynthesis utilizing the B. siamensis that can serve as a model for various commercial and industrial applications for efficient levan production.