Purification of pullulanase from Aureobasidium pullulans |
| |
| Authors: | Hani Moubasher Salwa S. Wahsh Nabil Abo El-Kassem |
| |
| Affiliation: | 1.Botany Department, Faculty of Science, Faculty of Science,Cairo University,Cairo,Egypt;2.Botany Department, Faculty of Science,Fayoum University,Fayoum,Egypt;3.Department of Botany, Faculty of Science,Fayoum University,Fayoum,Egypt |
| |
| Abstract: | Purification and characterization of pullulanase from Aureobasidium pullulans. Pullulanase was purified by using gel—filtration column then on ion exchange using Q-sepharose column yielding a single peak. Purification was further carried out on SP-sepharose column. Molecular weight of pullulanase from A. pullulans was found to be about 73 KDa on the SDS-PAGE 10%. Native-PAGE 10% showed the activity of pullulanase, using polyacrylamide gel containing pullulan. Hydrolysis products from pullulanase activity with soluble starch, glycogen and pullulan on thin layer chromatography appeared as one band which is maltotriose, while α-amylase with soluble starch and glycogen showed two bands which are maltose and maltotriose but α-amylase gave negative result with pullulan on TLC chromatography only. Pullulanase could degrade α-1,6 glycosidic linkage of the previous substrates, while amylase could degrade α-1,4 glycosidic linkage of glycogen, soluble starch and pullulan. MALDI-Ms was employed to deduce protein sequence of pullulanase. |
| |
| Keywords: | |
| 本文献已被 SpringerLink 等数据库收录！ |
|
