Method for determining activity of xylanase in forage

A technology of xylanase and determination method, which is applied in the direction of color/spectral characteristic measurement, etc., can solve the problems of low sensitivity, unsuitable enzyme quantitative analysis, unfavorable scientific research and product quality control, etc., and achieves small loss of activity, high sensitivity, The effect of easy operation

Active Publication Date: 2014-09-17
QINGDAO UNIV OF SCI & TECH +1
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Problems solved by technology

Xylanase belongs to polysaccharide hydrolase. At present, enterprises generally use DNS method to measure the activity of the enzyme. The country has also promulgated "GB-T 23874-2009 Spectrophotometric method for the determination of xylanase activity in feed additives". Its low sensitivity is difficult to meet the enzyme activity determination of the current compound feed with trace enzyme content, and it is only suitable for the determination of xylanase activity of a single feed additive
Although this method is simple and fast, its sensitivity is relatively low, and it is not suitable for the quantitative analysis of enzymes in feed, so it is not conducive to related scientific research and product quality control

Method used

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  • Method for determining activity of xylanase in forage
  • Method for determining activity of xylanase in forage

Examples

Experimental program
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Embodiment 1

[0026] Determination of xylanase activity in feed by test tube method

[0027] Reagents: xylan is beech xylan from sigma-aldrich; xylose is from sigma-aldrich; xylanase is from Wuhan xinhuayang biological Co., Ltd.; the compound feed containing xylanase comes from Qingdao Zhengda Feed Co., Ltd.; other reagents are of analytical grade; the water used in the experiment is distilled water.

[0028] 1 Endpoint method for determination of xylanase activity in feed

[0029] (1) Prepare 8mg / mL xylan solution: first dissolve 0.8000g xylan in water, dissolve it at 80°C, use a 50mL volumetric flask to set the volume to the mark, and then transfer the solution to a 100mL volumetric flask In the medium, use 0.1mol / L buffer solution to adjust the volume to the mark, mix thoroughly, and transfer to the reagent bottle for use;

[0030] (2) Preparation of 0.06mg / mL xylose standard solution: Accurately weigh 0.6000g of xylose dried to a constant mass, dissolve with 0.05mol / L acetic acid buff...

Embodiment 2

[0063] Determination of Xylanase Activity in Feed by Microplate Method

[0064] Reagents: xylan is beech xylan from sigma-aldrich; xylose is from sigma-aldrich; xylanase is from Wuhan xinhuayang biological Co., Ltd.; the compound feed was from Qingdao Liuhe Group; other reagents were of analytical grade; the water used in the experiment was distilled water.

[0065] (1) Prepare 8mg / mL xylan solution: first dissolve 0.8000g xylan in water, dissolve it at 80°C, use a 50mL volumetric flask to set the volume to the mark, and then transfer the solution to a 100mL volumetric flask In the medium, use 0.1mol / L buffer solution to adjust the volume to the mark, mix thoroughly, and transfer to the reagent bottle for use;

[0066] (2) Preparation of 0.12mg / mL xylose standard solution: Accurately weigh 1.0000g of xylose dried to a constant mass, dissolve with 0.05mol / L acetic acid buffer solution of pH5.5, transfer to 100mL, and make 10mg / mL mL of xylose standard solution; take 1.2mL of ...

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Abstract

The invention discloses a method for determining the activity of xylanase in forage, comprising the following steps: (1) drawing a standard curve in the corresponding relation between xylose optical density and xylose concentration; (2) preparing a leaching liquor of xylanase in the forage; (3) carrying out enzymatic hydrolysis; and (4) calculating the enzyme activity of xylanase in the forage according to the standard curve. According to the invention, the activities of the whole process of extracting xylanase in the forage get little loss, the loss is less than 10 %, high flux analysis can be realized, and the operation is simple and practical. The sensitivity of the method is obviously higher than that of a DNS method.

Description

technical field [0001] The invention relates to a method for measuring xylanase activity in feed, in particular to an enzyme extraction method in feed, specific measuring means and selection of key parameters for measuring. Background technique [0002] Xylanase can improve the utilization rate of animal feed, so it has been widely used in the feed industry. Enzyme activity is an important indicator to measure the biological activity of enzymes. Xylanase belongs to polysaccharide hydrolase. At present, enterprises generally use DNS method to measure the activity of the enzyme. The country has also promulgated "GB-T 23874-2009 Spectrophotometric method for the determination of xylanase activity in feed additives". Its low sensitivity has been difficult to meet the enzyme activity determination of the current compound feed with trace enzyme content, and it is only suitable for the determination of xylanase activity of a single feed additive. Although this method is simple an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/33
Inventor 张永勤王斐张杰常海燕詹志春张菁
Owner QINGDAO UNIV OF SCI & TECH
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