1,3-β-Glucanase from Vigna aconitifolia and its possible use in enzyme bioreactor fabrication

• Published in: International journal of biological macromolecules 2011-12, Vol.49 (5), p.894-899
• Main Authors: Kestwal, Rakesh Mohan, Bagal-Kestwal, Dipali, Chiang, Been Huang
• Format: Article
• Language: English
• Subjects: beta-glucanase; beta-glucans; beta-Glucans - metabolism; Bioreactors; Biotechnology - methods; calcium; Calcium - metabolism; Characterization; Chitosan - chemistry; Chitosan - metabolism; Chromatography, Gel; Chromatography, Ion Exchange; Circular Dichroism; Endo-1,3-beta-Glucanase - chemistry; Endo-1,3-beta-Glucanase - isolation & purification; Endo-1,3-beta-Glucanase - metabolism; Enzyme Stability; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - isolation & purification; Enzymes, Immobilized - metabolism; Fabaceae - chemistry; Fabaceae - enzymology; gel chromatography; glucose oxidase; Glucose Oxidase - chemistry; Glucose Oxidase - metabolism; glycoproteins; Hydrogen-Ion Concentration; Hydrolysis; Immobilization; immobilized enzymes; ion exchange chromatography; Kinetics; Plant Proteins - chemistry; Plant Proteins - isolation & purification; Plant Proteins - metabolism; Protein Structure, Secondary; Purification; Substrate Specificity; Vigna aconitifolia; β-Glucanase
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2011.08.002

Endo-1,3(4)-β-glucanase (EC 3.2.1.6) from Vigna aconitifolia sprouts was purified to 14.5 fold by gel filtration and ion-exchange chromatography. The enzyme was found to be a glycoprotein, its activity was Ca2+ dependent and specific for β-1,3 linkages in different polysaccharides. The Km value of the enzyme was estimated to be 3.0mgml−1 for β-d-glucan as substrate. Circular dichroism studies revealed 8% α-helix, 48% β-pleated and 44% random coil in its secondary structure. Purified β-glucanase was then successfully co-immobilized with glucose oxidase in agarose–chitosan beads, showing better immobilization yield, operational range and stability as compared with the crude β-glucanase beads. The immobilized β-glucanase was successfully used for mini-bioreactor fabrication.