title: 1,3-β-Glucanase from Vigna aconitifolia and its possible use in enzyme bioreactor fabrication.
abstract: 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 Ca(2+) dependent and specific for β-1,3 linkages in different polysaccharides. The K(m) value of the enzyme was estimated to be 3.0 mg ml(-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.
authors.affiliation: Institute of Food Science and Technology, National Taiwan University, No. 1, Roosevelt Road, Sec. 4, Taipei, Taiwan.
authors.forename: Rakesh Mohan
authors.initials: RM
authors.lastname: Kestwal
authors.forename: Dipali
authors.initials: D
authors.lastname: Bagal-Kestwal
authors.forename: Been Huang
authors.initials: BH
authors.lastname: Chiang
mesh_terms: D019149:Bioreactors; D001709:Biotechnology / Q000379:methods*; D002118:Calcium / Q000378:metabolism; D048271:Chitosan / Q000737:chemistry / Q000378:metabolism; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D002942:Circular Dichroism; D044644:Endo-1,3(4)-beta-Glucanase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D007887:Fabaceae / Q000737:chemistry / Q000201:enzymology*; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D017433:Protein Structure, Secondary; D013379:Substrate Specificity; D047071:beta-Glucans / Q000378:metabolism*
chemical_list: D004800:Enzymes, Immobilized; D010940:Plant Proteins; D047071:beta-Glucans; D048271:Chitosan; D005949:Glucose Oxidase; D044644:Endo-1,3(4)-beta-Glucanase; D002118:Calcium