Method of producing and purifying enzymes

Details Method of producing and purifying enzymes Method of producing and purifying enzymes

C - Chemistry, Metallurgy

12

N

C12N 9/00 (2006.01) C07K 19/00 (2006.01) C12N 9/38 (2006.01) C12N 9/96 (2006.01) C12N 11/16 (2006.01)

Patent

CA 2207278

A method of producing and purifying an enzyme which comprises selecting a spore forming host organism, preparing a genetic construct consisting of a DNA sequence encoding the desired enzyme and a DNA sequence directing synthesis of the desired enzyme during sporulation, inserting the genetic construct into the host organism, culturing the transformed host organism under sporulating conditions to obtain host organism spores with the enzyme integrally associated to the spores, and then treating the host organism and enzyme combination to remove any impurities, if necessary. The free enzyme can be obtained by cleaving the connection between the host organism and the enzyme. The combination of the enzyme and host organism is both a stabilized and an immobilized enzyme preparation.

Méthode de production et de purification d'une enzyme qui comprend la sélection d'un organisme hôte sporulé, la préparation d'une construction génétique consistant en une séquence d'ADN codant pour l'enzyme souhaitée et une séquence d'ADN dirigeant la synthèse de l'enzyme souhaitée durant la sporulation, l'insertion de la construction génétique dans l'organisme hôte, la culture l'organisme hôte transformé dans des conditions de sporulation afin d'obtenir des spores de cet organisme auxquels l'enzyme est intégralement associée, et le traitement par la suite de l'association organisme hôte et enzyme en vue d'en éliminer les impuretés, au besoin. L'enzyme libre peut être obtenue en coupant le lien entre l'organisme hôte et l'enzyme. L'association enzyme et organisme hôte est à la fois une préparation enzymatique stabilisée et immobilisée.

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