Towards the Identification of Type II Secretion Signals in a Nonacylated Variant of Pullulanase from Klebsiella oxytoca

• DOI: 10.1128/jb.187.20.7045-7055.2005
• Publication Date: 2005-09-30T16:23:42Z
• Authors: Olivera Francetić; Anthony P. Pugsley
• Keywords: Klebsiella oxytoca; Pectate lyase; Pullulanase

Pullulanase (PulA) from the gram-negative bacterium Klebsiella oxytoca is a 116-kDa surface-anchored lipoprotein of isoamylase family that allows growth on branched maltodextrin polymers. PulA specifically secreted via type II secretion system. PelBsp-PulA, nonacylated variant made by replacing signal peptide (sp) with pectate lyase PelB Erwinia chrysanthemi, was efficiently into medium. Two 80-amino-acid regions PulA, designated A and B, were previously shown to promote beta-lactamase (BlaM) endoglucanase CelZ fused C terminus. We show B can also BlaM but not nuclease NucB or several other reporter proteins. However, deletion most region all either individually together, had only minor effect PelBsp-PulA secretion. Four independent linker insertions between amino acids 234 324 in abolished This part C, could contain be important for its correct presentation. Deletion without dramatically affecting stability. PelBsp-PulA-NucB chimeras if PulA-NucB fusion point located downstream C. The data at least three information influences secretion, depending their context, some proteins might contribute which they are part.