Purification and characterization of a novel acid-tolerant and heterodimeric β-glucosidase from pumpkin (Cucurbita moschata) seed
| Autor: | Chang Woo Kwon, Eui Young Kim, Pahn-Shick Chang |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0301 basic medicine Circular dichroism Bioengineering 01 natural sciences Applied Microbiology and Biotechnology Substrate Specificity Gel permeation chromatography 03 medical and health sciences Cucurbita 010608 biotechnology Enzyme Stability Zymography Enzyme kinetics Protein secondary structure Chromatography Molecular mass biology Chemistry beta-Glucosidase Substrate (chemistry) Hydrogen-Ion Concentration biology.organism_classification Molecular Weight Kinetics 030104 developmental biology Cucurbita moschata Seeds Biotechnology |
| Zdroj: | Journal of Bioscience and Bioengineering. 132:125-131 |
| ISSN: | 1389-1723 |
| DOI: | 10.1016/j.jbiosc.2021.04.004 |
| Popis: | A novel β-glucosidase was purified from pumpkin (Cucurbita moschata) seed by anion exchange chromatography and gel permeation chromatography, and its molecular mass was determined to be 42.8 kDa by gel permeation chromatography. The heterodimeric structure consisting of two subunits, free from disulfide bonds, was determined by native-PAGE analysis followed by zymography. The enzyme was maximally active at pH 4.0 and 70°C, and Vmax, Km, and kcat values were 0.078 units mg−1 protein, 2.22 mM, and 13.29 min−1, respectively, employing p-nitrophenyl-β- d -glucopyranoside as the substrate. The high content of glycine determined by amino acid analysis implies that the enzyme possesses flexible conformations and interacts with cell membranes and walls in nature. Circular dichroism studies revealed that the high stability of the enzyme within the pH range of 2.0–10.0 is due to its reversible pH-responsive characteristics for α-helix–antiparallel β-sheet interconversion. |
| Databáze: | OpenAIRE |
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