Modified oligosaccharides used as substrate for measuring alpha-amylase activity

Claim 2

A process for using at least one modified oligosaccharide having at most 7 glucose units and having a substituent selected from the group consisting of 2-aminopyridyl, 3-aminopyridyl, anilino, methylanilino, hydroxyanilino, carboxyphenylamino, and hydroxyl groups at at least one end moiety of said oligosaccharide as substrate for measuring a-amylase activity or for measuring a-amylase isozymes.

Claim 3

A process according to Claim 1, wherein the modified oligosaccharide is represented by the formula: wherein R 1 is a 2-aminopyridyl a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylaminb group; and n is an integer of 2 to 5.

Claim 4

A process according to Claim 1, wherein the modified oligosaccharide is a mixture of and wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group.

Claim 5

A process according to Claim 1, wherein the modified oligosaccharide is represented by the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group.

Claim 6

A process according to Claim 2, wherein R 1 in the formula (I) is a 2-aminopyridyl group.

Claim 7

A process according to Claim 3, wherein the modified oligosaccharide is a mixture of

Claim 8

A process according to Claim 4, wherein the modified oligosaccharide is represented by the formula:

Claim 9

A process according to Claim 4, wherein the modified oligosaccharide is represented by the formula:

Claim 10

A process for measuring a-amylase activity which comprises incubating a sample using as substrate a modified oligosaccharide of the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and n is an integer of 2 to 5, and measuring the resulting product by high performance liquid chromatography.

Claim 11

A process for measuring a-amylase activity which comprises incubating a sample using as substrate a modified oligosaccharide of the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and n is an integer of 2 to 5, and measuring the resulting product by high performance liquid chromatography.

Claim 12

A process for measuring a-amylase activity which comprises incubating a sample using as substrate a modified oligosaccharide of the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and n is an integer of 2 to 5, and using as coupling enzyme a-glucosidase or glucoamylase to produce glucose, and measuring the amount of glucose by a spectrophotometric method.

Claim 13

A process for measuring a-amylase activity which comprises incubating a sample using as substrate a modified oligosaccharide of the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and n is an integer of 2 to 5, and using as coupling enzyme a-glucosidase or glucoamylase to produce glucose, and measuring the amount of glucose by a spectrophotometric method.

Claim 14

A process for measuring a-amylase isozymes which comprises conducting a transglycosylation reaction by using as substrate a mixture of modified oligosaccharides of the formulae: and wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group, by the action of a-amylase isozymes, conducting a hydrolysis reaction by usi-g as substrate the reaction product of the transglycosylation reaction between the modified oligosaccharides of the formulae (II) and (III), that is, the modified oligosaccharide of the formula: wherein R1 and R 2 are as defined above, by the action of a-amylase isozymes, and measuring the amount of a reaction product.

Claim 15

A process for measuring a-amylase isozymes which comprises conducting a transglycosylation reaction by using as substrate a mixture of modified oligosaccharides of the formulae: and wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group, by the action of a-amylase isozymes, conducting a hydrolysis reaction by usi-g as substrate the reaction product of the transglycosylation reaction between the modified oligosaccharides of the formulae (II) and (III), that is, the modified oligosaccharide of the formula: wherein R1 and R 2 are as defined above, by the action of a-amylase isozymes, and measuring the amount of a reaction product.

Claim 16

A process for measuring a-amylase isozymes derived from pancreas and derived from salivary glands which comprises conducting a hydrolysis reaction by using as substrate a modified oligosaccharide of the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group, by the action of a-amylase isozymes, and measuring the amount of a decomposed product by the hydrolysis.

Claim 17

A process for measuring a-amylase isozymes derived from pancreas and derived from salivary glands which comprises conducting a hydrolysis reaction by using as substrate a modified oligosaccharide of the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group, by the action of a-amylase isozymes, and measuring the amount of a decomposed product by the hydrolysis.

Claim 18

A modified oligosaccharide represented by the formula: wherein R1 is a 2 - am i no py r i d y l group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and n is an integer of 2 to 5.

Claim 19

A modified oligosaccharide represented by the formula: wherein R1 is a 2 - am i no py r i d y l group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and n is an integer of 2 to 5.

Claim 20

A mixture of modified oligosaccharides represented by the formulae: and wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group.

Claim 21

A mixture of modified oligosaccharides represented by the formulae: and wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group.

Claim 22

A modified oligosaccharide represented by the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group.

Claim 23

A modified oligosaccharide represented by the formula: wherein R 1 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group or a carboxyphenylamino group; and R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group, or a hydroxyl group.

Claim 24

A modified oligasaccharide represented by the formula: wherein R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group or a hydroxyl group.

Claim 25

A modified oligasaccharide represented by the formula: wherein R 2 is a 2-aminopyridyl group, a 3-aminopyridyl group, an anilino group, a methylanilino group, a hydroxyanilino group, a carboxyphenylamino group or a hydroxyl group.