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Application of laccase in decolorization of protein gel

A protein gel and laccase technology, which is applied in the application field of laccase in protein gel decolorization, can solve the problems that the study of dye wastewater decolorization is limited to aqueous environment, long decolorization time, and a large amount of waste liquid, etc., and achieves good decolorization. Effect, operation green environmental protection, energy saving effect

Inactive Publication Date: 2018-12-21
FUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

To obtain a better decolorization effect, a longer decolorization time (overnight) is required, and the decolorization solution must be replaced several times, which will produce a large amount of (organic) dye waste liquid, which will cause pollution if discharged directly
So far, studies on the decolorization of dye wastewater by laccases have been limited to aqueous environments

Method used

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  • Application of laccase in decolorization of protein gel
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  • Application of laccase in decolorization of protein gel

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Embodiment 1HYB07 laccase to the decolorization of different dyestuffs

[0044] (1) HYB07 laccase was used for 12 different dyes (Coomassie Brilliant Blue R250, Methyl Orange, Methyl Red, Basic Fuchsin, Bromothymol Blue, Neutral Red, Crystal Violet, Acid Fuchsin, Congo Red , bromocresol green, methylene blue, rose bengal B) decolorization. Decolorization system 2mL, containing 50mM citric acid-disodium hydrogen phosphate buffer (pH 6.0), dye (25-200mg / L) and enzyme (2U / mL), decolorize at room temperature for 24h. from figure 1 It can be seen that after laccase treatment, the decolorization rate of Coomassie Brilliant Blue is the highest, reaching 92.37%.

[0045] (2) Full-wavelength scanning of Coomassie Brilliant Blue R250 solution before and after laccase decolorization. from figure 2 It can be seen that the absorption peak of the dye decreases after decolorization.

Embodiment 2

[0046] Embodiment 2 utilizes laccase / mediator system to carry out the decolorization of protein gel

[0047] (1) Effect of Mediator Types on Protein Gel Decolorization

[0048] Under the condition of pH 7.0, 20U / mL laccase, detect different laccase mediators, including ACE (20μM), SYA (20μM), HBT (20μM), ABTS (2μM) and SYD (20μM) for protein gel decolorization Impact. from image 3 It can be seen that ABTS has the best effect as a mediator.

[0049] (2) Effect of temperature on protein gel decolorization

[0050] Under the conditions of pH 7.0, enzyme concentration 20U / mL, and ABTS 2μM, the effects of different temperatures (20°C, 25°C, 30°C, 35°C, 40°C, 45°C) on the decolorization of protein gels were studied. from Figure 4 It can be seen that 40°C is the optimum temperature for decolorization.

[0051] (3) Effect of laccase activity on protein gel decolorization

[0052] Under the conditions of pH 7.0 and temperature 40℃, the effect of laccase activity (3U / mL, 6U / mL,...

Embodiment 3

[0057] The detection sensitivity of embodiment 3 laccase / mediator decolorization system

[0058] (1) Decolorize for 2 hours under the conditions of pH 7.0, temperature 40°C, laccase activity 15U / mL, and ABTS 2μM, and detect the detection sensitivity of the laccase / mediator decolorization system. from Figure 8 It can be seen that BSA can still be detected as low as 10ng.

[0059] (2) Compare the effects of different decolorization methods. from Figure 9 It can be seen that the decolorization effect of the laccase / mediator decolorization system is the best, and there is little difference in the preparation of the decolorization solution with buffer or distilled water. Boiling with water directly for decolorization may easily cause loss of protein signal. Only water or buffer solution was decolorized for 2 hours at 40°C, and the background was high, indicating that laccase not only degrades R-250 in the solution, but also accelerates the decolorization of protein gel. Prot...

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Abstract

The invention belongs to the field of biotechnology, and more particularly relates to an application of laccase in decolorization of protein gel. Specifically, laccase is applied to decolorization after conventional dyeing in protein gel electrophoresis. The object of the invention is to use laccase to decolorize the dyed protein gel, which is environmentally-friendly, fast and simple, organic solvents are not required, energy is saved and the pollution of dye wastewater is reduced. According to the application of laccase in decolorization of protein gel, the protein gel is decolorized by laccase, the decolorization is rapid (clear protein band with low background may be obtained in 2 hours), the operation is simple (without boiling or replacement of decolorization liquid) and environmental (the organic solvents required by the traditional decolorization solution such as methanol or acetic acid are not used, dye degradation is achieved while decolorizing, and no waste disposal problemsand costs exist). A good decolorization effect is obtained in a short time by optimizing the laccase concentration, type and concentration of mediator, decolorization time and other factors, and theoperation is simple and green environmental.

Description

[0001] This application is a divisional application of the parent application with the application number 201510068601.0, the title of the invention is "Application of Laccase in Protein Gel Decolorization", and the filing date is February 10, 2015. technical field [0002] The invention belongs to the field of biotechnology, and more specifically relates to the application of laccase in protein gel decolorization. Background technique [0003] Laccase (laccase, EC 1.10.3.2) is a copper-containing polyphenol oxidase that widely exists in higher plants, fungi and insects, among which white-rot fungi are considered to be the most efficient laccase producers. Laccase is a green enzyme. Through oxidation reaction, laccase causes the substrate to lose electrons to form free radicals, and the lost electrons are transferred to molecular oxygen to form water. Laccase has a wide range of substrates and can catalyze the oxidation of various aromatic compounds, but the effect of laccas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/447
Inventor 杨捷杨晓丹林娟叶秀云
Owner FUZHOU UNIV
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