Isomaltodextrin-producing enzymes (PP enzyme) include two extracellular enzymes (α-glucosyltransferase and α-amylase) produced by Paenibacillus alginolyticus PP710. These enzymes are essential for producing isomaltodextrin (IMD), a highly branched α-glucan, from starch. In this study, we evaluated the safety of this PP enzyme. No genotoxicity was observed when the PP enzyme was assayed in standardized bacterial reverse mutation and chromosome aberration tests. An acute toxicity study in rats showed no toxic effects of PP enzyme at 2,000 mg/kg. No animals died, and no effects of enzyme administration were observed in a 14-day repeated oral-dose toxicity study in rats at the maximum dose of 1,000 mg/kg/day. The no observed adverse effect level was determined to be 1,000 mg/kg/day in a 90-day subchronic gavage toxicity study in rats. No animals died, and no abnormal findings due to consumption of the PP enzyme were observed in this study. These safety evaluation results demonstrated that the PP enzyme was safe for application as an ingredient in the manufacturing of the food ingredient IMD.