Highly Efficient Production of Transfructosylating Enzymes Using Low-Cost Sugarcane Molasses by A. Pullulans FRR 5284

Abstract In this study, sugarcane molasses was used to produce transfructosylating enzymes by A. pullulans FRR 5284. It was found that NaNO3 was a better nitrogen source than yeast extract while exogeneous phosphorous was not needed. Adding only 4.4 g/L NaNO3 into the molasses medium containing 100 g/L sugars led to the highest total transfructosylating activity of 123.8 U/mL. Scale-up of the enzyme production process from shake flasks to 1 L reactor improved the enzyme activity and productivity to 171.7 U/mL and 3.58 U/mL/h, 39% and 108% higher than the corresponding activity and productivity from shake flasks, respectively. FOS production from 500 g/L sucrose led to the highest yields of ~ 61% using intracellular, extracellular, and total enzymes from shake flasks and the reactor. Enzymes from different sources led to very different FOS profiles, indicating that FOS profiles can be controlled by adjusting intracellular and extracellular enzyme ratios to adjust prebiotic activity.