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Method for judging whether activity detection of glucoamylase is accurate or not

A technology for glucoamylase and activity detection, which is applied in the field of biochemical analysis, can solve problems such as large errors, and achieve the effect of overcoming excessive errors and satisfying accurate detection.

Pending Publication Date: 2020-12-22
JILIN COFCO BIOCHEM +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The invention discloses a method for judging whether the enzyme activity detection of glucoamylase is accurate, overcomes the problem of excessive error in the traditional glucoamylase measurement method, and provides a basis for detecting the accuracy of glucoamylase activity

Method used

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  • Method for judging whether activity detection of glucoamylase is accurate or not
  • Method for judging whether activity detection of glucoamylase is accurate or not
  • Method for judging whether activity detection of glucoamylase is accurate or not

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] 1) Use the national standard method to measure the activity of glucoamylase

[0047] ①Take 3 sample bottles and 1 blank control bottle, add 25mL starch solution with a mass concentration of 20g / L and 5mL acetic acid-sodium acetate buffer solution with a pH of 4.6 to each bottle, shake well in a 40°C water bath Preheat for 5-10 minutes;

[0048] ②Add 2mL of Genencor glucoamylase A to be tested to the 3 sample bottles respectively, shake well, and react in a water bath at 40°C for 30min;

[0049] ③Add 0.2mL NaOH solution with a mass concentration of 200g / L to 3 sample bottles respectively, shake well, and quickly cool with water to terminate the enzyme reaction; add 2mL Genencor glucoamylase A to the blank control bottle;

[0050] ④ Take 5mL of the reaction solution drawn from the sample bottle and the blank control bottle in step ③ into the iodine bottle, add 5mL of 0.1M iodine standard solution, and then add 15mL of 0.1M NaOH solution. The bottle was placed in the dar...

Embodiment 2

[0065] 1) Determination of the enzyme activity of glucoamylase by the national standard method

[0066] ①Take 3 sample bottles and 1 blank control bottle, add 25mL starch solution with a mass concentration of 20g / L and 5mL acetic acid-sodium acetate buffer solution with a pH of 4.6 to each bottle, shake well in a 40°C water bath Preheat for 5-10 minutes;

[0067] ②Add 2mL of BSJ glucoamylase to be tested to 3 sample bottles respectively, shake well, and react in 40℃ water bath for 30min;

[0068] ③Add 0.2mL NaOH solution with a mass concentration of 200g / L to 3 sample bottles respectively, shake well, and quickly cool with water to terminate the enzyme reaction; add 2mL BSJ glucoamylase to the blank control bottle;

[0069] ④ Take 5mL of the reaction solution drawn from the sample bottle and the blank control bottle in step ③ into the iodine bottle, add 5mL of 0.1M iodine standard solution, and then add 15mL of 0.1M NaOH solution. The bottle was placed in the dark to react f...

Embodiment 3

[0084] 1) Use the national standard method to measure the activity of glucoamylase

[0085] ① Take 3 sample bottles and 1 blank control bottle, add 25mL starch solution with a mass concentration of 20g / L and 5mL acetic acid-sodium acetate buffer solution with a pH of 4.6 to each bottle, shake well and pre- Heat for 5-10 minutes;

[0086] ②Add 2mL of Genencor glucoamylase B to be tested to the 3 sample bottles respectively, shake well, and react in a water bath at 40°C for 30min;

[0087] ③Add 0.2mL NaOH solution with a mass concentration of 200g / L to 3 sample bottles respectively, shake well, and quickly cool with water to terminate the enzyme reaction; add 2mL Genencor glucoamylase B to the blank control bottle;

[0088] ④ Take 5mL of the reaction solution drawn from the sample bottle and the blank control bottle in step ③ into the iodine bottle, add 5mL of 0.1M iodine standard solution, and then add 15mL of 0.1M NaOH solution. The bottle was placed in the dark to react for...

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Abstract

The invention discloses a method for judging whether activity detection of glucoamylase is accurate or not. A national standard method and a glucose kit method are combined, the enzyme activity measured by the two methods is divided to obtain a ratio, and the ratio is a fixed value; and in the process of producing and applying the glucoamylase, the two measuring methods are combined to obtain theenzyme activity ratio. When the activity of the glucoamylase is measured in industrial production, the accuracy of enzyme activity detection each time can be judged according to the ratio; and therefore, the requirement for accurately detecting the activity of the glucoamylase is met, and the problem that errors are too large in a traditional method for measuring the activity of the glucoamylase is solved.

Description

technical field [0001] The invention discloses a method for judging whether the detection of the activity of glucoamylase is accurate. The activity of the glucoamylase is determined by the combination of a national standard method and a glucose kit method, and belongs to the field of biochemical analysis. Background technique [0002] Glucoamylase (Glucoamylase EC3.2.1.3) is also called glucoamylase. It mainly exists in filamentous fungi and yeasts such as Aspergillus niger, Aspergillus oryzae and Rhizopus, and also exists in human saliva, animal pancreas and bacteria. There are 23 genera and 35 species of glucoamylase-producing fungal microorganisms reported, and 3 genera and 3 species of bacteria. The strains used in industrial production are mainly Aspergillus niger. Glucoamylase is an exoglycosidase, which sequentially hydrolyzes α-1,4 glycosidic bonds from the non-reducing end of starch to mainly generate glucose. Glucoamylase is widely used in the industrial producti...

Claims

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Application Information

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IPC IPC(8): C12Q1/40
CPCC12Q1/40G01N2333/934
Inventor 佟毅任晓冬李义陶进梁颖超李文钊李东旭张晓芸贾力耕
Owner JILIN COFCO BIOCHEM
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