Kanzaki Paper Mfg. Co., Ltd.

Minako Oka

Ryuzo Hayashi

Akio Kariyone

Yoshio Hashizume

Alcohol oxidase is immobilized by bonding an aminosilane coupling agent to a carrier, bonding a multifunctional aldehyde such as glutaraldehyde to an amino group of the aminosilane, washing the carrier to remove free multifunctional aldehyde and bonding alcohol oxidase to the multifunctional aldehyde bonded to the aminosilane. Washing to remove free multifunctional aldehyde enables immobilizing a one molecule thickness of alcohol oxidase on the carrier since free multifunctional aldehyde is not present to cross-link alcohol oxidase molecules together. A thin layer of alcohol oxidase is advantageous when assaying for alcohol since a thin layer does not retain hydrogen peroxide that can deactivate alcohol oxidase. The carrier preferably contains hydroxyl groups and is porous, and can be a silicate-containing carrier such as diatomaceous earth or fire brick. The immobilized alcohol oxidase is used in a column as part of an apparatus for measuring alcohol or as part of an apparatus containing immobilized glucose oxidase for measuring glucose and alcohol.

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Sapporo, Japan

Minako Oka