APPLICATION OF TRIS(HYDROXYMETHYL)PHOSPHINE AS A COUPLING AGENT FOR ALCOHOL-DEHYDROGENASE IMMOBILIZATION

The use of tris(hydroxymethyl)phosphine as a coupling reagent for the immobilization of alcohol dehydrogenase onto chitosan films and for th e attachment of the chitosan film to a glass support resulted in enzym e activities far above those obtained by adsorption of enzyme, and gre ater than those observed when using the more conventional glutaraldehy de-coupling protocol. The stability of the chitosan films was dramatic ally increased by covalent attachment to the glass using tris(hydroxym ethyl)phosphine. Similar enzyme activities were found for coupling wit h tris(hydroxymethyl)phosphine and glutaraldehyde on aminopropyl silic a and aminopropyl glass. The tris(hydroxymethyl)phosphine coupling ext ended the longevity of the alcohol dehydrogenase activity brit did not alter the pH optima or K-m of the enzyme.