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Lignin degradation liquid, preparation method and method for degrading lignin with it

A technology of lignin and degradation liquid, applied in the directions of biochemical equipment and methods, enzymes, enzymes, etc., can solve the problem of difficult to achieve high-efficiency biodegradation of lignin, and achieve the effect of improving the degradation conversion efficiency and the degradation efficiency.

Inactive Publication Date: 2019-01-04
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0005] The present invention provides a lignin degradation liquid, and at the same time provides its preparation method and a method for degrading lignin, so as to solve the problem that it is difficult to achieve high-efficiency biodegradation of lignin by using a single ligninase such as laccase

Method used

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  • Lignin degradation liquid, preparation method and method for degrading lignin with it
  • Lignin degradation liquid, preparation method and method for degrading lignin with it
  • Lignin degradation liquid, preparation method and method for degrading lignin with it

Examples

Experimental program
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Effect test

Embodiment 1

[0032] Example 1 Preparation of laccase LacT derived from Polyporus:

[0033] A. Draw the inoculum block from the slant culture of the polypore, inoculate it into 100ml of potato liquid seed medium, place it on a shaker and cultivate it at a speed of 200r / min for 8 days at a temperature of 25°C to obtain a liquid strain;

[0034] B. Insert the prepared liquid strain into the laccase-producing medium with an inoculum of 10% by volume, place the culture at 25° C. for static cultivation for 6 days, add 1 mM veratrol, and continue to cultivate respectively for 3, After 10 and 15 days, the culture was filtered to obtain crude laccase enzyme solution;

[0035] The preparation method of the laccase enzyme production medium is as follows: add 20g bran per 100ml Kirk nitrogen-limited liquid medium, and sterilize at 100°C for 40min.

[0036] C. Determination of laccase enzyme activity, the results are shown in Table 1, select the 3d laccase crude enzyme solution with the highest enzyme...

Embodiment 2

[0040] Example 2 Preparation of laccase LacG derived from Ganoderma lucidum:

[0041] A. Draw the inoculum piece from the slant culture of Ganoderma lucidum, inoculate it into 100ml potato liquid seed medium, place it on a shaker, shake it at a speed of 50r / min for 2 days, and obtain a liquid strain at a temperature of 37°C;

[0042] B. Insert the prepared liquid strain into the laccase enzyme production medium with an inoculum size of 50% by volume. Place the culture at 37°C for static culture for 2 days, add 4mM veratrol, continue the culture for 3, 10, and 15 days respectively, and filter the culture to obtain crude enzyme solution;

[0043] The preparation method of the laccase enzyme production medium is as follows: add 0.1g bran per 100ml Kirk nitrogen-limited liquid medium, and sterilize at 125°C for 10min.

[0044] C. Determination of laccase enzyme activity, the results are shown in Table 2, and the 15d laccase crude enzyme solution with the highest enzyme activity w...

Embodiment 3

[0048] Example 3 Preparation of laccase LacP derived from Pleurotus pachyrhiza:

[0049] A. Draw the inoculation block from the Pleurotus spp. culture, inoculate it into 100ml of potato liquid seed medium, place it on a shaker, shake it at a speed of 150r / min for 4 days, and obtain a liquid strain at a temperature of 28°C;

[0050] B. Insert the prepared liquid strain into the laccase enzyme production medium with an inoculum size of 20% by volume. After the culture was placed at 28°C for 4 days, 10mM veratrol was added, and the culture was continued for 3, 10, and 15 days respectively, and the culture was filtered to obtain a crude enzyme solution;

[0051] The preparation method of the laccase enzyme production medium is as follows: add 2g bran per 100ml Kirk nitrogen-limited liquid medium, and sterilize at 121°C for 30min.

[0052] C. Determination of laccase enzyme activity, the results are shown in Table 3, select the 10d laccase crude enzyme solution with the highest en...

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Abstract

The invention discloses a lignin degradation solution and a preparation method thereof as well as a method for degrading lignin by using the lignin degradation solution, and belongs to biochemical and bio-refinery methods to solve the problem that the efficient biodegradation of lignin is difficult to realize by adopting single ligninase of laccase at present. The lignin degradation solution is prepared by the following steps: dissolving laccase and manganese peroxidase into an acetic acid-sodium acetate buffer solution of which the pH value is 4-6 in an enzyme load ratio of (10:1)-(1:5) to ensure that the enzyme loads of the laccase and manganese peroxidase are respectively 1U / ml-50U / ml and 1U / ml-50U / ml; and then adding 1-10mM of MnSO4 and 0.1-1mM of H2O2, wherein the laccase and manganese peroxidase are obtained by fermenting white rot fungi to obtain extracellular crude enzyme liquid and then performing separation and purification respectively. According to the lignin degradation solution and the methods disclosed by the invention, the synergistic oxidative degradation of macromolecular lignin with rich structural diversity is realized, and compared with a degradation reaction system with single ligninase, the degradation rate of the macromolecules of the lignin can each 30-50%, the degradation conversion efficiency is significantly improved, and the degradation solution and the methods can be applied to the fields of bio-refinery of lignocelluloses, biological pulping or environmental treatment and the like.

Description

technical field [0001] The invention belongs to biochemical and biorefining methods, and in particular relates to a lignin degradation liquid, a preparation method and a method for degrading lignin with the same. Background technique [0002] With the continuous depletion of fossil resources, the use of biorefining technology to obtain liquid fuels such as ethanol and butanol from lignocellulosic raw materials, as well as chemical materials and chemicals will become an important supplement to petroleum refining. Lignin, which is highly cross-linked by a variety of phenylpropane-based structural units, is the main component of lignocellulose and one of the most abundant biopolymers on earth. The biotransformation of lignin is extremely difficult, which not only limits The conversion of polysaccharide components such as cellulose and hemicellulose in lignocellulose is also a major component of biorefinery waste. Therefore, the efficient degradation and conversion of lignin is...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N9/08C12P19/00
Inventor 余洪波孔雯倪浩翔张晓昱
Owner HUAZHONG UNIV OF SCI & TECH
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