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Xylanase activity measuring method

A technology of xylanase and determination method, which is applied in the field of selection of key parameters in the determination process, can solve problems such as low sensitivity, unfavorable research and product quality control, and achieve the effect of high sensitivity

Active Publication Date: 2012-05-23
WUHAN SUNHY BIOLOGICAL
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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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Embodiment 1

[0014] Main instruments: water bath constant temperature oscillator, SHZ-82 type, Guohua Electric Co., Ltd.; digital acidity meter, Shanghai Dapu Instrument Co., Ltd.; 721 visible light spectrophotometer, Shanghai Jinghua Technology Instrument Co., Ltd.; UV-2550 ultraviolet visible spectrophotometer Ji, SHIMADZU; xylan, derived from beechwood, sigma; xylose, sigma; xylanase (standard enzyme), Ningxia Hersbit Biotechnology Co., Ltd., Wuhan Xinhua Yang Biological Co., Ltd.

[0015] Solution configuration

[0016] MBTH Chromogenic Solution: 3mg / mL 3-methyl-2-benzothiazolone hydrazone solution and 1mg / mL dithiothreitol solution are mixed in equal amounts, ready to use and ready to use, effective within one day.

[0017] Xylose standard solution (0.032mg / mL): Accurately weigh 0.3200g of xylose dried to a constant mass, dissolve it with 50mM pH5.5 acetic acid buffer solution, transfer it to 100mL, and prepare 3.2mg / mL xylose Standard solution; take 1mL xylose solution from it, oper...

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

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/27
Inventor 张永勤王哲平
Owner WUHAN SUNHY BIOLOGICAL
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