Enzyme immobilization on ultrafine cellulose fibers via poly(acrylic acid) electrolyte grafts.
Ultrafine cellulose fiber (diameter 200-400 nm) surfaces were grafted with polyacrylic acid (PAA) via either ceric ion initiated polymerization or methacrylation of cellulose with methacrylate chloride (MACl) and subsequent free-radical polymerization of acrylic acid. PAA grafts by ceric ion initiated polymerization increased with increasing reaction time (2-24 h), monomer (0.3-2.4 M), and initiator (1-10 mM) concentrations, and spanned a broad range from 5.5-850%. PAA grafts on the methacrylated cellulose fibers also increased with increasing molar ratios of MACl to cellulosic hydroxyl groups (MACl/OH, 2-6.4) and monomer acrylic acid (AA) to initiator potassium persulfate (KPS) ratios ([AA]/[KPS], 1.5-6), and were in a much narrower range between 12.8% and 29.4%. The adsorption of lipase (at 1 mg/ml lipase and pH 7) and the activity of adsorbed lipase (pH 8.5, 30 degrees C), in both cases decreased with increasing PAA grafts. The highest adsorption and activity of the lipase on the ceric ion initiated grafted fibers were 1.28 g/g PAA and 4.3 U/mg lipase, respectively, at the lowest grafting level of 5.5% PAA, whereas they were 0.33 g/g PAA and 7.1 U/mg lipase, respectively, at 12.8% PAA grafts on the methacrylated and grafted fibers. The properties of the grafted fibers and the absorption behavior and activity of lipase suggest that the PAA grafts are gel-like by ceric-initiated reaction and brush-like by methacrylation and polymerization. The adsorbed lipase on the ceric ion-initiated grafted surface possessed greatly improved organic solvent stability over the crude lipase. The adsorbed lipases exhibited 0.5 and 0.3 of the initial activity in the second and third assay cycles, respectively.
Chen
Hong
H
Fiber and Polymer Science, University of California, Davis, CA 95616, USA.
Hsieh
You-Lo
YL
D000180:Acrylic Resins / Q000737:chemistry*; D002175:Candida / Q000201:enzymology; D002384:Catalysis; D002482:Cellulose / Q000737:chemistry*; D002563:Cerium / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D007477:Ions / Q000737:chemistry; D007700:Kinetics; D008049:Lipase / Q000737:chemistry; D012997:Solvents
D000180:Acrylic Resins; D004800:Enzymes, Immobilized; D007477:Ions; D012997:Solvents; D002563:Cerium; C006903:carbopol 940; D002482:Cellulose; D008049:Lipase