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Enzyme activity testing method for micro-biological degradation of benzene ring

A technology for microbial degradation and testing methods, which is applied in the field of enzyme activity testing for microbial degradation of benzene rings, can solve problems such as lack of universality, and achieve good universality and simple testing process

Inactive Publication Date: 2012-05-23
TIANJIN POLYTECHNIC UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the existing enzyme activity research methods, the enzyme activity is generally judged by examining the change of a certain side group chromophore, but due to the different chromophores of each substance, this kind of test method does not have good universality
Within the scope of the applicant's search, there is no report on the literature about directly adopting the benzene ring opening method to measure the enzyme activity

Method used

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  • Enzyme activity testing method for micro-biological degradation of benzene ring
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  • Enzyme activity testing method for micro-biological degradation of benzene ring

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The enzymatic activity of Bacillus F4 degrading terephthalic acid with terephthalic acid as the sole carbon source.

[0030] Use phosphate buffer (3g / L potassium dihydrogen phosphate and 7g / L disodium hydrogen phosphate) to prepare terephthalic acid solutions with concentrations of 1, 2, 5, 10, 20, 30, 40mg / L (standard solution), carry out full-wavelength scanning on the prepared terephthalic acid solution on an ultraviolet spectrophotometer, determine that its maximum absorption wavelength is at 240nm, and then measure the absorbance of the terephthalic acid solution with different concentrations at a wavelength of 240nm , plotted against concentration-absorbance (see figure 1 ), find out that the concentration range of its linear fitting degree reaches the standard (fitting degree reaches 0.99986) is 1-40mg / L, and the linear equation (2) is obtained by fitting: Y = 0.08342 +0.07584 ×X, where X represents terephthalate The concentration of formic acid, Y represents th...

Embodiment 2

[0035] Yeast powder was added as an organic nitrogen source, and the enzyme activity of Bacillus F4 degraded terephthalic acid.

[0036] The mensuration of terephthalic acid standard solution and the curve figure that corresponding concentration-absorbance draw are the same as embodiment 1 (see figure 1 ), fitting the linear equation (2): Y = 0.08342 +0.07584 ×X, where X represents the concentration of terephthalic acid, Y represents the UV absorbance, the linear equation (2) can be used to calculate the benzene ring content in the degradation process of this example The change.

[0037]Adding yeast powder as an organic nitrogen source in the catalytic reaction solution can effectively promote the consumption rate of the carbon source by microorganisms, thereby increasing the enzyme activity. In order to determine the degree of improvement of enzyme activity, 1g / L yeast extract powder was added to the reaction solution, and 3g / L potassium dihydrogen phosphate and 7g / L disodiu...

Embodiment 3

[0042] Enzyme activity of bacteria TJ degrading protocatechuic acid with protocatechuic acid as the sole carbon source.

[0043] Use phosphate buffer (4g / L potassium dihydrogen phosphate and 6g / L disodium hydrogen phosphate) to prepare protocatechuic acid solutions with concentrations of 1, 2, 5, 10, 20, 30, 40mg / L Wavelength scanning, determine the maximum absorption wavelength at 250nm, measure the absorbance of solutions with different concentrations at the wavelength of 250nm, and make a curve corresponding to the concentration-absorbance (see figure 2 ), find out that the concentration range of its linear fitting degree reaches the standard (fitting degree reaches 0.99971) is 1-40mg / L, and the linear equation (3) is obtained by fitting: Y = 0.07613 +0.07579 × X, where X represents protocatechu Acid concentration, Y represents the UV absorbance, and the linear equation (3) can be used to calculate the change of benzene ring content during the degradation process.

[0044...

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Abstract

The invention discloses an enzyme activity testing method for micro-biological degradation of a benzene ring. The testing method comprises the following steps of: 1, making a standard curve of relation between substrate concentration and absorbance, and finding out a linear fitting equation between the substrate concentration and the absorbance; 2, preparing substrate solution according to enzymeor bacterial suspension of which the activity is to be measured; 3, previously heating the substrate solution and the enzyme solution to the most proper reaction temperature respectively, mixing the substrate solution and the enzyme solution, performing catalytic reaction on the mixture under the most proper reaction condition, and detecting the variation in the substrate concentration after the reaction; 4, measuring benzene ring content of the reaction solution, namely diluting the reaction solution before and after the catalytic reaction to ensure that the absorbance of the reaction solution is within the range of the linear equation, measuring the absorbance at the maximum absorption wavelength, and calculating the substrate concentration before and after the reaction according to thelinear equation; and 5, calculating the enzyme activity U for the micro-biological degradation of the benzene ring according to a formula (1) of U=delta C / M / delta t*103.

Description

technical field [0001] The invention relates to a technology for testing enzyme activity in the catalysis process of whole cells of microorganisms, in particular to a method for testing enzyme activity of microorganisms degrading benzene rings. Background technique [0002] Substances containing benzene rings are widely used in the chemical industry, so the amount of industrial wastewater formed is large, and benzene ring substances have certain toxic effects on the environment, and even have carcinogenic and teratogenic effects on organisms. Therefore, benzene ring substances degradation is a hot topic of research. Biodegradation is a commonly used method for the degradation of pollutants. Benzene rings have certain biodegradability. Therefore, there are many studies on microbial degradation of substances containing benzene rings. [0003] During the degradation process of substances containing benzene rings, the opening of benzene rings is an important indicator for evalu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/75C12Q1/25
Inventor 张健飞张卫玲李政巩继贤李秋瑾
Owner TIANJIN POLYTECHNIC UNIV