Pullulanase (EC 3.2.1.41, limit dextrinase, amylopectin 6-glucanohydrolase, bacterial debranching enzyme, debranching enzyme, α-dextrin endo-1,6-α-glucosidase, R-enzyme, pullulan α-1,6-glucanohydrolase) is a specific kind of glucanase, an amylolytic exoenzyme, that degrades pullulan.

[1][2][3] It is produced as an extracellular, cell surface-anchored lipoprotein by Gram-negative bacteria of the genus Klebsiella.

Its substrate, pullulan, is regarded as a chain of maltotriose units linked by α-1,6-glycosidic bonds.

Pullulanase will hydrolytically cleave pullulan (α-glucan polysaccharides).

It is additionally used in low-calorie food formulations as a starch replacement.