Activation of polyvinyl chloride sheet surface for covalent immobilization of oxalate oxidase and its evaluation as inert support in urinary oxalate determination.
Polyvinyl chloride (PVC) sheets are a promising material for enzyme immobilization owing to the PVC's properties such as being chemically inert, corrosion free, weather resistant, tough, lightweight, and maintenance free and having a high strength-to-weight ratio. In this study, this attractive material surface was chemically modified and exploited for covalent immobilization of oxalate oxidase using glutaraldehyde as a coupling agent. The enzyme was immobilized on activated PVC surface with a conjugation yield of 360 microg/cm(2). The scanning electron micrographs showed the microstructures on the PVC sheet surface revealing the successful immobilization of oxalate oxidase. A colorimetric method was adopted in evaluating enzymatic activity of immobilized and native oxalate oxidase. The immobilized enzyme retained 65% of specific activity of free enzyme. Slight changes were observed in the optimal pH, incubation temperature, and time for maximum activity of immobilized oxalate oxidase. PVC support showed no interference when immobilized oxalate oxidase was used for estimation of oxalic acid concentration in urine samples and showed a correlation of 0.998 with the values estimated with a commercially available Sigma kit. The overall results strengthen our view that PVC sheet can be used as a solid support for immobilization of enzymes and in the field of clinical diagnostics, environmental monitoring and remediation.
Pundir
Chandra Shekhar
CS
Biochemistry Research Laboratory, Department of Biochemistry and Genetics, MD University, Rohtak, 124001 Haryana, India. pundircs@yahoo.com
Chauhan
Nar Singh
NS
Bhambi
Manu
M
D001681:Biological Assay; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D010070:Oxalates / Q000378:metabolism / Q000652:urine*; D010088:Oxidoreductases / Q000737:chemistry / Q000378:metabolism*; D011143:Polyvinyl Chloride / Q000737:chemistry*; D013499:Surface Properties; D013997:Time Factors
D004800:Enzymes, Immobilized; D010070:Oxalates; D011143:Polyvinyl Chloride; D010088:Oxidoreductases; C020598:oxalate oxidase