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Double-enzyme method for improving specificity of gamma-cyclodextrin

A cyclodextrin, specific technology, applied in the direction of fermentation, etc., can solve the problem of low specificity of γ-cyclodextrin, and achieve the effect of improving the specificity and increasing the catalytic temperature

Active Publication Date: 2015-06-24
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The invention solves the problem of low specificity of γ-cyclodextrin, and further expands its application range

Method used

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  • Double-enzyme method for improving specificity of gamma-cyclodextrin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Dissolve cornstarch in 50ml of Tris-Hcl buffer solution with a pH of 8.5 and a concentration of 50mmol / l to make a 1% solution, and heat it with microwave for 2-3min to gelatinize it.

[0024] (2) Take 1ml of the solution prepared in (1), first add 0.1U / g starch 4αGTase to react at 70°C for 24h to pretreat corn starch, and then add 0.5U / g starch CGTase to the reaction system at 60 Reaction at ℃ for 24h.

[0025] (3) Boil the reaction solution in step (2) to inactivate it for 30 minutes.

[0026] (4) Mix 500 ul of the reaction solution in step (3) with 500 ul of 0.2 M sodium acetate buffer solution at pH 4.2, and add 0.4 U / g starch glucoamylase to it and react at 38° C. for 16 h.

[0027] (5) Boil the reaction solution obtained in step (4) for 10 minutes to inactivate the enzyme, and centrifuge at 18000 g for 5 minutes after inactivating the enzyme.

[0028] (6) The cyclodextrin solution measured in the above steps was measured, and the ratio of γ-cyclodextrin / β-cy...

Embodiment 2

[0030] (1) Dissolve cornstarch in 50ml of Tris-Hcl buffer solution with a pH of 8.5 and a concentration of 50mmol / l to make a 1% solution, and heat it with microwave for 2-3min to gelatinize it.

[0031] (2) Take 1ml of the solution prepared in (1), first add 0.1U / g starch 4αGTase to react at 70°C for 24h to pretreat corn starch, and then add 0.5U / g starch CGTase to the reaction system at 70°C Under reaction 24h.

[0032] (3) Boil the reaction solution in step (2) to inactivate it for 30 minutes.

[0033] (4) Mix 500 ul of the reaction solution in step (3) with 500 ul of 0.2 M sodium acetate buffer solution at pH 4.2, and add 0.4 U / g starch glucoamylase to it and react at 38° C. for 16 h.

[0034] (5) Boil the reaction solution obtained in step (4) for 10 minutes to inactivate the enzyme, and centrifuge at 18000 g for 5 minutes after inactivating the enzyme.

[0035] (6) The cyclodextrin solution measured in the above steps was measured, and the ratio of γ-cyclodextrin / β-cyc...

Embodiment 3

[0037] (1) Dissolve cornstarch in 50ml of Tris-Hcl buffer solution with a pH of 8.5 and a concentration of 50mmol / l to make a 1% solution, and heat it with microwave for 2-3min to gelatinize it.

[0038] (2) Take 1ml of the solution prepared in (1), first add 0.05U / g starch 4αGTase to react at 70°C for 24h to pretreat corn starch, and then add 0.5U / g starch CGTase to the reaction system at 70 Reaction at ℃ for 24h.

[0039] (3) Boil the reaction solution in step (2) to inactivate it for 30 minutes.

[0040] (4) Mix 500 ul of the reaction solution in step (3) with 500 ul of 0.2 M sodium acetate buffer solution at pH 4.2, and add 0.4 U / g starch glucoamylase to it and react at 38° C. for 16 h.

[0041] (5) Boil the reaction solution obtained in step (4) for 10 minutes to inactivate the enzyme, and centrifuge at 18000 g for 5 minutes after inactivating the enzyme.

[0042] (6) The cyclodextrin solution measured in the above steps was measured, and the ratio of γ-cyclodextrin / β-c...

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Abstract

A double-enzyme method for improving specificity of gamma-cyclodextrin includes the steps: (1) taking corn starch as a substrate, and subjecting the corn starch to microwave heating until the corn starch is completely dissolved into a Tris-Hcl buffer solution with pH being 8.5 and concentration being 50mmol / L to obtain corn starch solution; (2) respectively adding two enzymes including 4alpha GTase and CGTase into the corn starch solution, controlling reaction temperature and reaction time, and boiling for 30min to inactivate the enzymes after reaction is finished; (3) adding glucamylase into reaction solution, reacting for 16-24h at the temperature of 38 DEG C, and after reaction is finished, boiling for 10min to inactivate the enzymes and centrifuging for 5min at a speed of 10000rpm; (4) obtaining supernatant which is a mixture containing gamma-cyclodextrin. By the double-enzyme method for improving specificity of gamma-cyclodextrin, the problem of low specificity of gamma-cyclodextrin is solved, and application range of gamma-cyclodextrin can be further broadened.

Description

technical field [0001] The invention relates to the technical field of food, in particular to a method for improving the specificity of gamma-cyclodextrin by adopting a double-enzyme method. Background technique [0002] Cyclodextrins are a type of cyclic oligosaccharides produced by the action of glucosyltransferases produced by certain species of Bacillus on starch. The most notable feature of cyclodextrin is that it has a stereochiral cavity with a certain size, which is hydrophilic outside the ring and hydrophobic inside the ring, so it can embed some hydrophobic molecules or groups. γ-cyclodextrin is composed of 8 D-glucopyranose units linked by α-1,4-glycosidic bonds. Compared with α and β cyclodextrins, γ-cyclodextrins have a larger hydrophobic cavity, higher solubility, and can include larger molecules, so they have a wider application space in the food and medical industries. [0003] However, so far, β-cyclodextrin and to a lesser extent α-cyclodextrin have been ...

Claims

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Application Information

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IPC IPC(8): C12P19/20C12P19/18C12P19/04
Inventor 王金鹏李林林张梦柯周星赵建伟杨哪谢正军徐学明金征宇焦爱权田耀旗吴凤凤
Owner JIANGNAN UNIV
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