Covalent immobilization of pullulanase on alginate and study of its hydrolysis of pullulan.

The immobilization of pullulanase from Klebsiella pneumoniae by grafting was investigated. Pullulanase was linked after activation of alginate via a covalent bond between the amine groups of the enzyme and the carboxylic acid groups of alginate. The immobilization yield was 60%.

Molecular size characterization and kinetics studies on hydrolysis of pullulan by pullulanase in an entangled alginate medium.

The behavior of a hydrolytic enzyme (pullulanase) toward its substrate (pullulan) in the presence of a nonsubstrate (alginate), both below and above the critical entanglement concentration (C*), was studied. The hydrolysis kinetics were studied with the enzyme and alginate concentrations varied using two main methods: a colorimetric assay of the reducing extremities (RE), which allowed the number-average molar masses (Mn) of the oligosaccharides to be determined, and size exclusion chromatography with on-line, multiangle light scattering, viscometer, and differential refractive index detectors, which allowed the average molar masses, Mn and Mw, of the oligosaccharides during hydrolysis to be determined. Free pullulanase acts via an "endo" process.

Effect of carboxymethyl groups on degradation of modified pullulan by pullulanase from Klebsiella pneumoniae.

Pullulanase is an enzyme that hydrolyses the α-1,6 linkages of pullulan (Pull) to produce maltotriose units. We studied the capacity of pullulanase to cleave its modified substrate: carboxymethylpullulan (CMPull), synthesized with two different degrees of substitution (DS=0.16 and 0.