A protease-resistant α-galactosidase characterized by relatively acid pH tolerance from the Shitake Mushroom Lentinula edodes
| Autor: | Bowen Chen, Ming-chang Chang, Cui-ping Feng, Xue-ran Geng, Jun-long Meng, Xu Lijing |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
medicine.medical_treatment
Shiitake Mushrooms 02 engineering and technology Biochemistry Stachyose Substrate Specificity 03 medical and health sciences chemistry.chemical_compound Structural Biology Endopeptidases Enzyme Stability medicine Food science Amino Acid Sequence Raffinose Melibiose Molecular Biology 030304 developmental biology Ions 0303 health sciences Protease Guar gum biology Hydrolysis Temperature General Medicine Hydrogen-Ion Concentration 021001 nanoscience & nanotechnology biology.organism_classification Enzyme Activation Molecular Weight Papain Kinetics Lentinula chemistry Metals alpha-Galactosidase Locust bean gum 0210 nano-technology Chromatography Liquid |
| Zdroj: | International journal of biological macromolecules. 128 |
| ISSN: | 1879-0003 |
| Popis: | A monomeric α-galactosidase with a molecular weight of 64 kDa was purified from fresh fruiting bodies of Lentinula edodes. The purification protocol involved ion-exchange chromatography on DEAE-cellulose, CM-cellulose and Q-Sepharose and a final gel-filtration on Superdex 75. The purified α-galactosidase (LEGI) was identified by LC-MS/MS. It demonstrated the optimum pH of 5.0 and temperature optimum of 60 °C towards pNPGal. It was inhibited by Cd2+, Fe3+, Pb2+, Zn2+, Al3+, Hg2+, Cr2+, Ba2+. The LEGI activity was strongly abolished by the chemical modification N-bromosuccinimide (NBS) at 1 mM, while significantly enhanced by the thiol-reducing agents dithiothreitol (DTT). Moreover, LEGI showed strong resistance to protease pepsin, papain, acid protease and neutral protease. LEGI demonstrated hydrolysis towards melibiose (13.27%), raffinose (4.75%), stachyose (2.58%), locust bean gum (0.82%) and guar gum (1.29%). The Km values of LEGI for pNPGal, stachyose, raffinose, and melibiose were found to be 1.08, 17.24, 13.80 and 8.05 mM, respectively. Results suggest that LEGI demonstrates potential for elimination of indigestible oligosaccharides. |
| Databáze: | OpenAIRE |
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