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Electrochemical determination method for peroxidase activity in compost

A technology of peroxidase and manganese peroxidase, which is applied in the field of determination of peroxidase activity, can solve the problems of being affected by light-interfering substances, high turbidity of compost leachate, and low accuracy

Inactive Publication Date: 2007-09-05
HUNAN UNIV
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AI Technical Summary

Problems solved by technology

[0004] At present, spectrophotometry is generally used for the determination of LiP and MnP activity, such as the veratrol method for determining LiP activity and the use of certain dyes (RB brilliant blue, aniline blue, azure, etc.) as indicators of LiP activity. The limitation of substrate turbidity and the influence of light interfering substances; while the KMB (2-keto-4-methiolbutyric acid)-ethylene analysis method is slow, and there are many oxidants besides LiP that can oxidize KMB to produce ethylene, and the selectivity is not good
The commonly used phenol red method, divalent manganese oxidation spectrophotometry, and guaiacol method to determine MnP activity also have strict requirements on the turbidity of the substrate, and are affected by light-interfering substances, so the accuracy of the measurement is not high.
Excessive turbidity of compost leachate hampers the application of spectrophotometric determination of peroxidase activity

Method used

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  • Electrochemical determination method for peroxidase activity in compost
  • Electrochemical determination method for peroxidase activity in compost
  • Electrochemical determination method for peroxidase activity in compost

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Embodiment Construction

[0035] 1. Pretreatment of glassy carbon electrodes

[0036] With 0.5μm Al 2 O 3 The powder is polished until the surface of the electrode is smooth, and then the surface of the electrode is rinsed with water, followed by 1:1 HNO. 3 , acetone, water ultrasonic cleaning, and finally rinse with water, then dry naturally.

[0037] 2. Bacteria culture

[0038] P. chrysosporium was inoculated into PDA medium and cultured at 37°C for 4 days for bacterial expansion. The spores were inoculated into Kirk liquid medium, cultured with shaking at a rate of 120 r / min at 37° C. for 8 days, and the crude enzyme liquid was obtained by filtration.

[0039] PDA medium composition: potato 40g, agar 4g, glucose 4g, water 200ml.

[0040] Kirk liquid medium composition: glucose 10g / L, ammonium tartrate 0.2g / L, Tween 80 1g / L, benzyl alcohol 0.54g / L, KH 2 PO 4 2.56g / L, MgSO 4 0.71g / L, VB1 0.001g / L, phthalic acid buffer 10mmol / L, trace element solution 70ml / L; final pH 4.5.

[0041] Among them...

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Abstract

This invention relates to an electrochemistry measurement of the peroxid enzyme activity, the method uses the auricome and eldopaque as the enzyme base and measures the activity of the hyperoxide in the dunghill lixivium with xanthan even bacterium and establishes the linear relation between the current change slope and the enzyme activity. The relation is as following: its peroxide enzyme is 2.27-29.79U / L and manganese peroxide is 0.085-1.37U / L.

Description

Technical field: [0001] The invention relates to a method for measuring peroxidase activity, in particular to the electrochemical measurement of peroxidase activity in a complex composting system. Background technique: [0002] Phanerochaete chrysosporium is a white-rot fungus that plays an important role in degrading lignin and various organic pollutants in municipal solid waste composting. The lignin-degrading enzyme system formed by the bacteria when some main nutrients (such as nitrogen, carbon, sulfur) are limited has a unique ability to degrade various xenobiotics, it can degrade lignin in wood and make wood white , Studies since the 1980s have shown that the bacteria have non-specific characteristics of substrate oxidation and can degrade a variety of organic substances, including natural polymers, reduced compounds, oxidized compounds, and toxic compounds. There are two main enzymes in the lignin-degrading enzyme system synthesized by P. chrysosporium: lignin peroxi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26G01N27/416
Inventor 汤琳曾光明黄国和章毅
Owner HUNAN UNIV
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