Relationship between the induced-fit loop and the activity of Klebsiella pneumoniae pullulanase

Abstract

Klebsiella pneumoniae pullulanase (KPP) belongs to glycoside hydrolase family 13 subfamily 13 (GH13_13) and is the only enzyme that is reported to perform an induced‐fit motion of the active‐site loop (residues 706–710). Comparison of pullulanase structures indicated that only KPP has Leu680 present behind the loop, in contrast to the glycine found in other GH13_13 members. Analysis of the structure and activity of recombinant pullulanase from K. pneumoniae ATCC 9621 (rKPP) and its mutant (rKPP‐G680L) indicated that the side chain of residue 680 is important for the induced‐fit motion of the loop 706–710 and alters the binding affinity of the substrate.