Method for quantitative determination of 1,5-anhydroglucitol

C - Chemistry – Metallurgy – 12 – N

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C12N 9/24 (2006.01) C12Q 1/34 (2006.01) C12Q 1/48 (2006.01)

Patent

CA 2218488

The present invention relates to a method for quantitative determination of l, 5-anhydroglucitol in a sample, which comprises mixing the sample and an enzyme having activity that is inhibited by 1, 5-anhydroglucitol in a concentration-dependent manner, and measuring the activity of the enzyme; and a reagent for quantitative determination of l, 5-anhydroglucitol which comprises an enzyme having activity that is inhibited by 1, 5-anhydroglucitol in a concentration-dependent manner, a substrate for the enzyme, and a reagent for quantitative determination of a product formed by the enzyme activity. The present invention also relates to novel trehalase having a Ki value of 0.33 mM or less for 1, 5-anhydroglucitol; and a process for producing novel trehalase having the above-mentioned physicochemical properties, which comprises culturing in a medium a microorganism belonging to the genus Nocardia and capable of producing the trehalase, and recovering the trehalase from the culture.

Méthode de détermination du 1,5-anhydroglucitol se caractérisant par le fait que l'on ajoute ce 1,5-anhydroglucitol contenu dans un échantillon à un enzyme dont l'activité est inhibée par le glucitol d'une manière dépendante de la concentration et que l'on analyse l'activité de l'enzyme ainsi inhibé. L'invention porte également sur un réactif permettant de déterminer le 1,5-anhydroglucitol comprenant un enzyme dont l'activité est inhibée par le glucitol d'une manière dépendante de la concentration, un substrat de l'enzyme, et un réactif permettant de déterminer une substance générée par l'activité de cet enzyme. L'invention porte enfin sur une nouvelle tréhalase présentant une valeur Ki de 0,33 mM ou moins par rapport au 1,5-anhydroglucitol et sur un procédé de production d'une nouvelle tréhalase ayant les propriétés physico-chimiques précitées, se caractérisant par la culture de souches appartenant au genre Nocardia capable de produire la tréhalase dans un milieu de culture et sur la récolte de la tréhalase ainsi produite dans cette culture.

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