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Method for measuring carboxymethylcellulose enzyme activity

A technology of carboxymethyl cellulose and sodium carboxymethyl cellulose, which is applied in the field of determination of carboxymethyl cellulose enzyme activity, can solve problems such as errors, and achieve the effects of small errors, good reproducibility, and fast test speed

Inactive Publication Date: 2009-02-04
DONGGUAN BAOLIMEI CHEM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The national industry standard detection method is based on the IPUAC method, and major changes have been made in the establishment of the substrate, the dilution factor of the enzyme solution, and the production of glucose marking lines. The time required for the detection of samples by this method is similar to the IPUAC recommended method, and Due to the difference of domestic substrates, it brings greater error

Method used

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  • Method for measuring carboxymethylcellulose enzyme activity
  • Method for measuring carboxymethylcellulose enzyme activity
  • Method for measuring carboxymethylcellulose enzyme activity

Examples

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Effect test

example 1

[0031] Add 200μL of the enzyme solution to be tested diluted to an appropriate multiple into a 2mL disposable plastic centrifuge tube, then add 1% sodium carboxymethylcellulose (Sigma, low viscosity) solution substrate equal in volume to the enzyme solution, and mix well. Place the reaction at 50℃ for 10 minutes; at the same time as a blank control of the enzyme, add 200μL of the enzyme solution to be tested diluted to an appropriate multiple into the reaction tube, and then add 3 times the volume of the enzyme solution DNS (3,5-dinitrosalicylic acid) Reagents, mix well, and react at 50℃ for 10 minutes; take out all reaction tubes, immediately add 3 times the volume of enzyme solution DNS (3,5-dinitrosalicylic acid) reagent to the sample tube, mix well, and add the enzyme to the control tube Liquid equal volume of 1% sodium carboxymethylcellulose (Sigma, low viscosity 4%) solution substrate, mix well; put all reaction tubes in a boiling water bath and heat for 5 minutes, take them...

example 2

[0037] Add 100μL of the enzyme solution to be tested diluted to an appropriate multiple into a 2mL disposable plastic centrifuge tube, then add 1% sodium carboxymethylcellulose (Sigma, low viscosity) solution substrate equal in volume to the enzyme solution, and mix well. Place the reaction at 50℃ for 10 minutes; at the same time as a blank control of the enzyme, add 100μL of the enzyme solution to be tested diluted to an appropriate multiple into the reaction tube, and then add 3 times the volume of the enzyme solution DNS (3,5-dinitrosalicylic acid) Reagents, mix well, and react at 50℃ for 10 minutes; take out all reaction tubes, immediately add 3 times the volume of enzyme solution DNS (3,5-dinitrosalicylic acid) reagent to the sample tube, mix well, and add the enzyme to the control tube Liquid equal volume of 1% sodium carboxymethylcellulose (Sigma, low viscosity) solution substrate, mix well; put all reaction tubes in a boiling water bath and heat for 5 minutes, take them ou...

example 3

[0042] Add 300μL of the enzyme solution to be tested diluted to an appropriate multiple into a 2mL disposable plastic centrifuge tube, then add 1% sodium carboxymethylcellulose (Sigma, low viscosity) solution substrate equal in volume to the enzyme solution, and mix well. Put it at 50℃ for 10min; at the same time as a blank control of the enzyme, add 300μL of the enzyme solution to be tested diluted to an appropriate multiple into the reaction tube, and then add 3 times the volume of the enzyme solution DNS (3,5-dinitrosalicylic acid) Reagents, mix well, and react at 50℃ for 10min; take out all reaction tubes, immediately add 3 times the volume of enzyme solution DNS (3,5-dinitrosalicylic acid) reagent to the sample tube, mix well, and add the enzyme to the control tube Liquid equal volume of 1% sodium carboxymethylcellulose (Sigma, low viscosity) solution substrate, mix well; put all reaction tubes in a boiling water bath and heat for 5 minutes, take them out, and quickly cool to...

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Abstract

The invention discloses a determination method for the activity of carboxymethyl cellulose. Acidic or neutral cellulose enzyme solution to be tested is firstly prepared: liquid enzyme is diluted by acidic or neutral buffer solution; the enzyme solution to be tested is added in a reaction tube, is added into sodium carboxymethylcellulose solution substrate with equivalent volume, DNS (3, 5-dinitrosalicylic acid) reagent with three times volume being enzymatic blank control, and is mixed evenly and reacts under the temperature of 50 DEG C; next all the reaction tubes are taken out, the DNS (3, 5- dinitrosalicylic acid) reagent with three times volume is added into the sample tube, sodium carboxymethylcellulose solution substrate with equivalent volume is added in a control tube; then all the reaction tubes are heated by water bath for 5 minutes and taken out to be cooled to the room temperature, and the reaction is ceased; finally, the reacted solution is taken to carry out the colorimetric determination of the absorbance value under the wavelength of 540nm, and the reducing sugar content is calculated according to the glucose standard curve. Compared with the prior art, the determination method has small variation coefficient of many tests for a same sample, good reproducibility of test results and fast test speed, and is suitable for determining numerous samples simultaneously.

Description

Technical field [0001] The invention relates to a method for measuring cellulase activity, in particular to a method for measuring carboxymethyl cellulase activity (CMCNa-DNS). Background technique [0002] Cellulase (Cellulase) is a multi-component complex enzyme. Cellulase from different sources differs greatly in composition and component ratio. Generally speaking, cellulases include exoβ-1,4-glucanase (Exop-1,4-glucanase, E C3.2.1.91), endoβ-1,4-glucanase ( Endop-1, 4-glucanase, E C3.2.1.4) and cellobiase (Cellobiase, E C3.2.1.21). Cellulase products from different sources have different ratios of the content of the three enzyme components, so their final apparent enzyme activity will be different. At the same time, the substrates that cellulase acts on are also relatively complicated, resulting in many methods for determining cellulase activity, and the methods are complex and not uniform. Commonly used methods are: carboxymethyl cellulose saccharification force method, carb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/34
Inventor 朱桂芳袁文杰李芬芳张盛圳黄碧莲
Owner DONGGUAN BAOLIMEI CHEM
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