Xylanase activity measuring method

A technology of xylanase and assay method, applied in the field of selection of key parameters in the assay process, can solve the problems of low sensitivity, unfavorable research and product quality control, etc., and achieve the effect of high sensitivity

Active Publication Date: 2009-07-08
QINGDAO UNIV OF SCI & TECH
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AI Technical Summary

Problems solved by technology

Although this method is simple and fast, its sensitivity is relatively low, which is not conducive to some scientific research and product quality control

Method used

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Experimental program
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Embodiment 1

[0014] Main instruments: water bath constant temperature oscillator, SHZ-82, Guohua Electric Co., Ltd.; digital acidity meter, Shanghai Dapu Instrument Co., Ltd.; 721 Visible Spectrophotometer, Shanghai Jinghua Technology Instrument Co., Ltd.; Ji, SHIMADZU; xylan, derived from beechwood, sigma; xylose, sigma; xylanase (standard enzyme), Ningxia Heshibi Biotechnology Co., Ltd., Wuhan Xinhuayang Biological Co., Ltd.

[0015] Solution configuration

[0016] MBTH chromogenic solution: 3-methyl-2-benzothiazolone hydrazone solution of 3 mg / mL and dithiothreitol solution of 1 mg / mL are mixed in equal amounts.

[0017] Standard xylose solution (0.032mg / mL): Accurately weigh 0.3200g xylose dried to constant mass, dissolve, transfer and dilute to 100mL with 50mM acetate buffer solution with pH 5.5 to prepare 3.2mg / mL xylose Standard solution; take 1 mL of xylose solution from it, operate as before and dilute to 100 mL to obtain 0.032 mg / mL xylose standard solution.

[0018] The prepar...

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PUM

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Abstract

The invention discloses a method for determining the activity of xylanase, which is characterized by comprising: A, producing a standard corresponding relation curve between an absorbance value of xylose measured by an MBTH method and concentration of the xylose; and B, using the MBTH method to detect the absorbance value which is produced after the xylanase to be tested hydrolyzes xylan and is equivalent to that of the xylose; and determining the content of reducing sugar which is produced in unit time and is equivalent to the xylose in an enzymolysis product of the xylanase to be tested according to the standard corresponding relation curve which is produced in step A between the concentration of the xylose and the measured absorbance value of the xylose to calculate the activity of the xylanase. The method uses the MBTH method to detect the quantity of terminal groups of the xylose produced after the xylanase hydrolyzes the xylan to determine the activity of the xylanase. The detectability and work concentration range of xylose determination by the method are apparently lower than those of a DNS method and a potassium ferricyanide method, so the sensitivity of the method is apparently higher than that of the DNS method and the potassium ferricyanide method.

Description

technical field [0001] The present invention relates to a method for measuring xylanase activity, in particular to a method for measuring xylanase activity by MBTH method, in particular to the selection of key parameters in the measuring process. Background technique [0002] Xylanases are hydrolases that degrade xylan into xylo-oligosaccharides and xylose. In recent years, because of its great application value in paper industry, food industry, feed industry, energy industry and environmental science, it has attracted extensive attention of scientific researchers, and related research has been carried out. Enzyme activity is an important indicator to measure the biological activity of enzymes. Xylanase is a polysaccharide hydrolase. Currently, the DNS method is commonly used to determine its enzyme activity. The specific method is to take 2 mL of xylose solution with a certain concentration gradient into a test tube, add 1.5-2.5 mL of DNS reagent, mix well, and place in a ...

Claims

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Application Information

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IPC IPC(8): G01N21/27
Inventor 张永勤王哲平
Owner QINGDAO UNIV OF SCI & TECH
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