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Method for degradation of non-phenolic lignin

A lignin, non-phenolic technology, applied in biochemical equipment and methods, enzymes, oxidoreductases, etc., can solve the problem of inability to directly degrade non-phenolic structural units, achieve a wide range of applications, improve degradation efficiency, cost low investment

Active Publication Date: 2017-09-05
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the low redox potential of manganese peroxidase, it can only oxidize lignin with phenolic structure, and cannot directly degrade the most abundant non-phenolic structural unit in lignin

Method used

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  • Method for degradation of non-phenolic lignin
  • Method for degradation of non-phenolic lignin
  • Method for degradation of non-phenolic lignin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] The preparation of embodiment 1 recombinant manganese peroxidase

[0021] The amino acid sequence of MnP-1 is as follows:

[0022] MAFKTILAFVALATAALAAPSSRVTCSPGRVVSNGACCKWFDVLDDIQENLFDGGVCGEEVHESLRLTFHDAIGFSLSAEREGKFGGGGADGSIMAFAEIETNFHANNGVDEIVEAQRPFAIKHKVSFGDFIQFAGAVGVSNCLGGPRLEFMAGRSNISRAAPDLTVPEPSDSVDKILARMGDAGFSSSEVVDLLISHTVAAQDHVDPTIPGTPFDSTPSEFDPQFFVETLLKGTLFPGNGSNVGELQSPLRGEFRLQSDALLARDPRTACEWQSFVNNQRLMVTKFEAVMSKLAVLGHNPRDLVDCSEVIPVPPRAKTNVAVLPAGKTRADVQAACAATPFPTLQTAPGPATSIVPV

[0023] The amino acid sequence of MnP-2 is as follows:

[0024] MAFKHLVVALSIVLSLGVAQAAITKRVACPDGKNTATNAACCSLFAIRDDIQANLFDGGECGEEVHESFRLTFHDAIGTGSFGGGGADGSIIVFDDIETNFHANNGVDEIIDEQKPFIARHNITPGDFIQFAGAVGVSNCPGAPRLDFFLGRPNPVAAAPDKTVPEPFDTVDSILARFKDAGGFTPAEIVALLGSHTIAAADHVDPTIPGTPFDSTPEVFDTQVFVEVQLRGTLFPGTGGNQGEVQSPLRGEIRLQSDHDLARDSRTACEWQSFVNNQAKLQSAFKAAFKKLSVLGHNINNLIDCSEVIPEPPNVKVKPATFPAGITHADVEQACATTPFPTLATDPGPATSVAPVPPS

[0025] Get Escherichia coli engineering strains producing mangane...

Embodiment 2

[0026] Example 2 Degradation of non-phenolic lignin by recombinant manganese peroxidase

[0027] The following reaction system: 50 μl malonic acid buffer (0.2M, pH 5.0), 20 μl deionized water, 20 μl non-phenolic lignin veratrol solution, 5 μl manganese sulfate (40 mM), 100 μl manganese peroxidase (1000 U / L), 5 μl hydrogen peroxide (4 mM). The system without adding manganese peroxidase was used as a control, and each manganese peroxidase was replicated three times. The reaction was degraded for 48h at 30°C, and the degradation rate of veratrol was analyzed by high performance liquid chromatography (HPLC). For specific experimental results, see figure 1 .

[0028] Such as figure 1 It was shown that manganese ions are an essential part of the recombinant manganese peroxidase to degrade non-phenolic lignin. In the absence of manganese ions, neither the recombinant peroxidases MnP-1 nor MnP-2 could degrade non-phenolic lignin. prime model compounds.

Embodiment 3

[0029] Example 3 Degradation of non-phenolic lignin by recombinant manganese peroxidase in different organic acid systems

[0030] According to the following reaction system: 50 μl organic acid buffer (0.2M, pH 5.0), 20 μl deionized water, 20 μl veratrol solution, 5 μl manganese sulfate (40 mM), 100 μl manganese peroxidase (1000U / L), 5 μl peroxide Hydrogen (4mM). The organic acid buffers used were malonic acid-sodium malonate, oxalic acid-sodium oxalate, acetic acid-sodium acetate, citric acid-sodium citrate, lactic acid-sodium lactate and succinic acid-sodium succinate. Different organic acid systems of manganese peroxidase were used as controls, and three replicates were set for each manganese peroxidase. The reaction was degraded for 48h at 30°C, and the degradation rate of veratrol was analyzed by high performance liquid chromatography (HPLC). For specific experimental results, see figure 2 .

[0031] Such as figure 2 and image 3 It was shown that different organic...

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Abstract

The invention relates to the technical field of biology, in particular to a method for degradation of non-phenolic lignin. Degradation efficiency of manganese peroxidase is improved by adding organic acid as an interbody during degradation of the non-phenolic lignin. Experimental results show that a non-phenolic lignin compound such as veratryl alcohol can be effectively degraded by addition of malonic acid and oxalic acid. The method is simple in operation, low in cost investment, wide in application, free of pollution, obvious in degradation efficiency of the non-phenolic lignin and widely applicable to the fields of pretreatment of lignocellulosic biomass and degradation of xenobiotics.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for degrading non-phenolic lignin. Background technique [0002] As a new type of renewable resource, lignocellulosic biomass is widely used in the fields of bioenergy and biobased chemicals. Lignocellulose is composed of cellulose, hemicellulose and lignin. Cellulose and hemicellulose are composed of basic structural units of sugar units. Lignin is mainly composed of a small amount of phenolic and a large number of non-phenolic substructural units. In the process of enzymatic saccharification of lignocellulose, lignin not only serves as a physical barrier to limit the access of cellulase to cellulose, but also adsorbs cellulase, resulting in non-reactive binding. Therefore, the content and type of lignin have a great influence on the hydrolysis efficiency of various lignocellulosic raw materials. [0003] Manganese peroxidase is a heme-containing glycosylation peroxidase,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/08
CPCC12N9/0065C12Y111/01013
Inventor 姚斌苏小运秦星罗会颖黄火清柏映国王苑王亚茹马锐涂涛马建爽
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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