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Alkaline xylanase activity determination method

A technique for measuring xylanase and activity, which is applied in the field of alkaline xylanase activity determination, and can solve the problem of decreased activity of alkaline xylanase, deviation of alkaline xylanase activity, and the inability to objectively react to alkali sexual xylanase and other issues

Inactive Publication Date: 2015-07-29
SOUTH CHINA UNIV OF TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The activity of alkaline xylanase under acidic conditions is not optimal and will cause a decrease in the activity of alkaline xylanase, thus indicating a large deviation in the activity of alkaline xylanase
Traditional assay methods can no longer objectively reflect the activity of alkaline xylanase, and new methods are urgently needed

Method used

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  • Alkaline xylanase activity determination method
  • Alkaline xylanase activity determination method
  • Alkaline xylanase activity determination method

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Embodiment 1

[0033] The determination of the activity of alkaline xylanase produced by Trichoderma reesei ATCC56765 culture and fermentation, the steps are as follows:

[0034] (1) Centrifuge the crude enzyme liquid produced by the cultivation and fermentation of Trichoderma reesei ATCC56765 at 8000r / min for 5min, and take the alkaline xylanase supernatant to be tested;

[0035] Weigh 1.000 g of anhydrous xylose, add sodium dihydrogen phosphate-disodium hydrogen phosphate buffer solution with pH=7.8 to dissolve, and set the volume to 100 mL to obtain xylose standard solution;

[0036] At 50°C, weigh 1.0000g of xylan, add 0.32g of sodium hydroxide, then add 90mL of water, stir magnetically, and heat at the same time until the xylan is completely dissolved; then stop heating, continue stirring for 30min, and add 1.18g of phosphoric acid Disodium hydrogen phosphate; continue magnetic stirring, and measure its pH at the same time; if the pH is 7.80, use sodium dihydrogen phosphate-disodium hyd...

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Abstract

The invention discloses an alkaline xylanase activity determination method and belongs to the field of xylanase activity determination. The method comprises the following steps: preparing a sodium biphosphate-sodium hydrogen phosphate buffer solution with the pH value of 7.8, and on the basis, preparing a xylose standard solution, a xylan standard solution and an alkaline xylanase sample; creating a xylose absorbency-xylose concentration standard curve; in the alkaline environment with the pH value of 7.8, using alkaline xylanase to be detected to hydrolyze xylan, and detecting the absorbency AE of the xylose obtained through hydrolysis; meanwhile, providing a blank control group not added with alkaline xylanase and determining the absorbency AB; correspondingly acquire the content of hydrolysis product xylose according to the measured absorbency of xylose obtained through hydrolysis by use of a standard curve; calculating the alkaline xylanase activity according to a formula. The alkaline xylanase activity determination method is improved to solve the problem that a conventional detection method cannot be used for objectively reflecting the activity of alkaline xylanase.

Description

technical field [0001] The invention belongs to the field of assaying xylanase activity, in particular to a method for assaying alkaline xylanase activity. Background technique [0002] Xylanases are hydrolases that degrade xylan into monosaccharides and oligosaccharides. For a long time, it has a relatively significant application value in papermaking, food, feed, energy and other industries, so it has attracted extensive attention of scientific researchers and has carried out related research on it. Enzyme activity is an important indicator to measure the biological activity of enzymes. For the determination of xylanase activity, there is an established national standard GB 23874-2009 Determination of xylanase activity of feed additives, but this standard is determined at 37°C and pH 5.50. For many industries, the alkaline environment makes the use of xylanase not very convenient. With the research and development of xylanase, new types of alkaline xylanase are emerging...

Claims

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Application Information

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IPC IPC(8): G01N21/31G01N1/28
Inventor 马千里杨仁党杨飞阮蒙
Owner SOUTH CHINA UNIV OF TECH
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