Fructooligosaccharides and levan have a wide range of applications in the food industry due to their physiological and functional properties. The enzymatic synthesis of these molecules exhibits great advantages when compared with microbial fermentation. In this study, the production of levansucrase from Bacillus subtilis natto and its utilization in fructooligosaccharides and levan syntheses using different reaction conditions were described. The best condition for levansucrase production was 420.7 g L-1 of sucrose at pH 7.0, which reached 23.9 U ml-1 of transfructosylation activity. In a bioreactor, the highest production of fructooligosaccharides was 41.3 g L-1 using a medium containing 350 g L-1 sucrose at 35 °C for 36 h. The enzymatic synthesis of levan resulted in 86.9 g L-1 when conditions similar to those used for fructooligosaccharides synthesis were applied. These results indicate that the levansucrase from B. subtilis natto could be applied for the co-production of fructooligosaccharides and levan, which are biomolecules that have health benefits and are used successfully in the food industry.