Immobilization and stabilization of invertase on Cajanus cajan lectin support.
Use of lectins as ligands for the immobilization and stabilization of glycoenzymes has immense application in enzyme research and industry. But their widespread use could be limited by the high cost of their production. In the present study preparation of a novel and inexpensive lectin support for use in the immobilization of glycoenzymes containing mannose or glucose residues in their carbohydrate moiety has been described. Cajanus cajan lectin (CCL) coupled covalently to cyanogen bromide activated Seralose 4B could readily bind enzymes such as invertase, glucoamylase and glucose oxidase. The immobilized and glutaraldehyde crosslinked preparations of invertase exhibited high resistance to inactivation upon exposure to enhanced temperature, pH, denaturants and proteolysis. Binding of invertase to CCL-Seralose was however found to be readily reversible in the presence of 1.0 M methyl alpha-D mannopyranoside. In a laboratory scale column reactor the CCL-Seralose bound invertase was stable for a month and retained more than 80% of its initial activity even after 60 days of storage at 4 degrees C. CCL-Seralose bound invertase exhibited marked stability towards temperature, pH changes and denaturants suggesting its potential to be used as an excellent support for the immobilization of other glycoenzymes as well.
Ahmad
S
S
Department of Biochemistry, Faculty of Life Sciences and Institute of Biotechnology, Aligarh Muslim University, Uttar Pradesh, India. ahmads@njc.org
Anwar
A
A
Saleemuddin
M
M
D019149:Bioreactors; D001798:Blood Proteins / Q000378:metabolism; D002241:Carbohydrates / Q000737:chemistry*; D003488:Cyanogen Bromide / Q000378:metabolism; D004795:Enzyme Stability / Q000187:drug effects*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005087:Glucan 1,4-alpha-Glucosidase / Q000378:metabolism; D005949:Glucose Oxidase / Q000378:metabolism; D005976:Glutaral / Q000494:pharmacology; D006026:Glycoside Hydrolases / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D037102:Lectins / Q000737:chemistry* / Q000191:economics / Q000378:metabolism; D008024:Ligands; D008766:Methylmannosides / Q000378:metabolism; D037121:Plant Lectins; D010944:Plants; D011509:Proteoglycans; D012995:Solubility; D013696:Temperature; D013997:Time Factors; D014508:Urea / Q000494:pharmacology; D043324:beta-Fructofuranosidase
D001798:Blood Proteins; D002241:Carbohydrates; D004800:Enzymes, Immobilized; D037102:Lectins; D008024:Ligands; D008766:Methylmannosides; D037121:Plant Lectins; D011509:Proteoglycans; C042057:proteolysis-inducing peptide; C008466:methylmannoside; D014508:Urea; D005949:Glucose Oxidase; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D005087:Glucan 1,4-alpha-Glucosidase; D003488:Cyanogen Bromide; D005976:Glutaral