Beta-mannase for decoloration and application of beta-mannase

A mannanase and application technology, applied in the directions of glycosylase, enzyme, hydrolase, etc., can solve the problems of toxic by-products, high energy consumption and high cost

Inactive Publication Date: 2016-06-01
HEILONGJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional physical and chemical methods are limited in practical application d...

Method used

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  • Beta-mannase for decoloration and application of beta-mannase
  • Beta-mannase for decoloration and application of beta-mannase
  • Beta-mannase for decoloration and application of beta-mannase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1. Obtaining of β-mannanase

[0019] Transfer the overnight cultured B. licheniformis HDGLJT-01 seed solution to konjac gum fermentation enzyme production medium, shake and culture at 37°C and 160rpm for 48h, take 40mL of fermentation broth, centrifuge at 4500r / min, 4°C for 20min, and obtain the supernatant , the crude enzyme solution. The crude enzyme liquid was purified by acetone precipitation method, and further purified by ion exchange chromatography and gel filtration chromatography. Specifically, the concentrated enzyme solution was added to a well-balanced DEAE-sepharoseFF ion exchange column, and 0.1mol / L NaCl and 0.005mol / L Tris-HCl with a pH of 7.0 were used for gradient elution, and the eluate was collected. Detection of enzyme activity and protein content. Add 5mL of ion-exchange-purified enzyme solution to a well-balanced sepadexG-75 column, use pure water as the eluent, and the elution flow rate is 0.5mol / min, collect the eluate, and detect th...

Embodiment 2

[0020] Example 2. Partial property analysis of β-mannanase

[0021] The activity analysis of the β-mannanase of the present invention includes the optimum temperature and thermostability of the enzyme and the optimum pH and pH stability of the enzyme.

[0022] (1) Optimum temperature and thermostability of β-mannanase

[0023] Take 2mL Tris-HCl buffer solution (0.1mol / L, pH 8.0) and mix it with 0.5mL pure enzyme solution. After incubation for 30min, immediately measure the enzyme activity. Enzyme activity is used as a control (this enzyme activity is the original enzyme activity, ie 100%), and the enzyme activity retention rate at different temperatures is calculated to determine the optimum temperature of β-mannanase. the result shows( figure 2 ), the optimal temperature of the enzyme is 60°C, indicating that the enzyme is more heat-resistant.

[0024] Mix 2mL of Tris-HCl buffer solution with 0.5mL of pure enzyme solution at 30°C, 40°C, 50°C, 60°C, 70°C and 80°C, and me...

Embodiment 3

[0028] The decolorization effect of embodiment 3.β-mannanase

[0029] Table 1 The dyes used in the decolorization test

[0030]

[0031]

[0032] Eight dyes with different structures (Table 1) were selected to detect the decolorization ability of β-mannanase. A certain amount of β-mannanase and dye were mixed, 37°C, 160rpm shaking reaction, at 0, 6 and 12h, full-wavelength scanning was performed to detect the residual situation of the dye. the result shows( Figure 7 ), β-mannanase has the best decolorization effect on azo dyes, among which methyl orange, Congo red and Dardan yellow are the best in decolorization. 50%, better decolorization. The decolorization effect of triaryl methane dyes is better, among which the decolorization rate of water-soluble aniline blue, rubenite green and phenol red is about 90%, and the effect is obvious; the decolorization rate of crystal violet is more than 50%, and the decolorization effect is better.

[0033] The decolorization e...

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Abstract

The invention relates to an application of beta-mannase in dye decoloration. Dye is azo dye and triarylmethane dye, wherein the azo dye is congo red, methyl orange, titan yellow and eriochrome black T; and the triarylmethane dye is water-soluble aniline blue, malachite green, crystal violet and phenol red. The beta-mannase is produced by bacillus licheniformis and preferably by bacillus licheniformis HDGLJT-01. The beta-mannase is prepared specifically by the method comprising the following steps: transferring and inoculating a bacillus licheiformis HDGLJT-01 seed solution cultured overnight to a konjac glucomannan fermented enzyme production culture medium, oscillating and culturing for 48 hours at a speed of 160 rpm at 37 DEG C, acquiring 40mL of fermentation liquor, and centrifuging for 20 minutes at a speed of 4500 r/min at 4 DEG C to obtain supernatant, namely, a crude enzyme solution; and purifying the crude enzyme solution by utilizing an acetone precipitation method, and further purifying by adopting ion-exchange column chromatography and gel filtration chromatography. Compared with the commercialized beta-mannase, the decoloration effect of the product is better.

Description

technical field [0001] The invention relates to the application of a mannanase, in particular to the decolorization application of the enzyme. Background technique [0002] Dyestuffs are widely used in industries such as food, pharmaceuticals, cosmetics, textiles and leather. At present, there are more than 10,000 kinds of dyes reported, and the annual production of dyes exceeds 70 million tons. In the process of dye production and use, a large amount of dye wastewater is directly discharged into the environment, which not only affects the transparency and dissolved oxygen of the water body, but also produces carcinogens such as benzidine. Therefore, the treatment of dye wastewater has become more and more popular. s concern. Traditional physical and chemical methods are limited in practical application due to their high cost, high energy consumption, and easy formation of toxic by-products. In this case, there is an urgent need for a treatment method with low cost and ene...

Claims

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Application Information

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IPC IPC(8): C02F3/34C12N9/24
CPCC02F3/342C02F2101/308C12N9/2491C12Y302/01025
Inventor 葛菁萍平文祥赵丹李兴霖金曼
Owner HEILONGJIANG UNIV
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