Purification and properties of the catalytic domain of the thermostable pullulanase type II from Thermococcus hydrothermalis

Authors
Erra-Pujada, M Chang-Pi-Hin, F Debeire, P Duchiron, F O'Donohue, MJ
Citation
M. Erra-pujada et al., Purification and properties of the catalytic domain of the thermostable pullulanase type II from Thermococcus hydrothermalis, BIOTECH LET, 23(16), 2001, pp. 1273-1277
Citations number
16
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
BIOTECHNOLOGY LETTERS
ISSN journal
01415492 → ACNP
Volume
23
Issue
16
Year of publication
2001
Pages
1273 - 1277
Database
ISI
SICI code
0141-5492(200108)23:16<1273:PAPOTC>2.0.ZU;2-Z
Abstract
A pullulanase type II was produced in Escherichia coli using the relevant g ene from Thermococcus hydrothermalis. This protein was purified and its pul lulanolytic and amylolytic activities were characterised. The optimum tempe rature and Ca2+ concentration for each activity were identical (105 degrees C and 0.09 mM), whereas the optimum pH (pH(pullulan) 5.75, pH(amylose) 5) a nd the influence of Ca2+ ions on the kinetic parameters were different. Fur ther analyses revealed that this enzyme exhibits an endo-processive-like ac tion and specifically cleaves alpha -1,6 bonds in pullulan.