title: Steady-state generation of hydrogen peroxide: kinetics and stability of alcohol oxidase immobilized on nanoporous alumina.
abstract: Alcohol oxidase from Pichia pastoris was immobilized on nanoporous aluminium oxide membranes by silanization and activation by carbonyldiimidazole to create a flow-through enzyme reactor. Kinetic analysis of the hydrogen peroxide generation was carried out for a number of alcohols using a subsequent reaction with horseradish peroxidase and ABTS. The activity data for the immobilized enzyme showed a general similarity with literature data in solution, and the reactor could generate 80 mmol H2O2/h per litre reactor volume. Horseradish peroxidase was immobilized by the same technique to construct bienzymatic modular reactors. These were used in both single pass mode and circulating mode. Pulsed injections of methanol resulted in a linear relation between response and concentration, allowing quantitative concentration measurement. The immobilized alcohol oxidase retained 58 % of initial activity after 3 weeks of storage and repeated use.
authors.affiliation: Department of Chemistry-BMC, Uppsala University, Husargatan 3, Box 576, 75123 Uppsala, Sweden. marcus.kjellander@kemi.uu.se
authors.forename: Marcus
authors.initials: M
authors.lastname: Kjellander
authors.forename: Kathrin
authors.initials: K
authors.lastname: Götz
authors.forename: Josefine
authors.initials: J
authors.lastname: Liljeruhm
authors.forename: Mats
authors.initials: M
authors.lastname: Boman
authors.forename: Gunnar
authors.initials: G
authors.lastname: Johansson
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D000438:Alcohols / Q000378:metabolism; D000537:Aluminum Oxide; D019149:Bioreactors; D004800:Enzymes, Immobilized / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D007700:Kinetics; D010843:Pichia / Q000201:enzymology*
chemical_list: D000438:Alcohols; D004800:Enzymes, Immobilized; D006861:Hydrogen Peroxide; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D000537:Aluminum Oxide