A marine sediment-derived lignin-degrading bacteria and its application in degrading lignin

A technology for lignin degrading bacteria and marine sediments, which is applied in the field of applied microorganisms to achieve the effects of strong lignin degradation ability, increased separation efficiency and strong application value

Active Publication Date: 2022-07-12
ZHEJIANG WANLI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Based on this, it is speculated that the lignin-rich marine sediment environment must be rich in lignin-degrading bacteria. However, there are few research reports on lignin-degrading bacteria from marine sediment environments.

Method used

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  • A marine sediment-derived lignin-degrading bacteria and its application in degrading lignin
  • A marine sediment-derived lignin-degrading bacteria and its application in degrading lignin
  • A marine sediment-derived lignin-degrading bacteria and its application in degrading lignin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Acquisition of lignin-degrading bacteria DBX666

[0020] (1) Enrichment culture of lignin-degrading bacteria

[0021] Add 5 g of sediment to a conical flask containing 100 mL of enrichment medium, and cultivate at 8°C at 150 rpm for 15 days to obtain the first-generation enriched culture; 1% of the inoculum was inoculated into an Erlenmeyer flask with fresh enrichment medium, and cultured at 8°C at 150 rpm for 14 days to obtain a second-generation enriched culture; The % inoculum was inoculated into a conical flask with enriched medium, and cultured at 8°C at 150 rpm for 14 days to obtain the third-generation enriched culture. The sediments were from a station in the East Pacific. Sediment samples, the enriched medium formula is (g / L): K 2 HPO 4 , 0.1g; KH 2 PO 4 , 0.4g; (NH 4 ) 2 SO 4 , 0.7125g; KNO 3 , 0.63125g; yeast extract, 0.1g; dealkalized lignin, 1g. Adjust the pH of the medium to 7.0.

[0022] (2) Screening and purification of lignin-degrad...

Embodiment 2

[0024] Example 2 Molecular identification and naming preservation of lignin-degrading bacteria DBX666 strain

[0025] (1) Molecular identification of strains

[0026]The conventional molecular cloning method was used to amplify and determine the 16S rDNA gene of the strain DBX666. The operation steps were as follows: ① DNA template preparation: pick a single colony with an inoculation loop into a 0.2 mL PCR tube containing 100 μl sterile water, boil at 98 °C for 15 min, Obtain a DNA template for PCR amplification. ②PCR amplification: The 16S rDNA amplification system (50ul) of the strain includes 0.5 μl of high-fidelity Taq enzyme, 5 μl of 10×Buffer, 4 μl of dNTP, 1 μl of upstream primer (1492R), 1 μl of downstream primer (27F), 1 μl of DNA template, deionized 37.5 μl of water; PCP reaction conditions are: pre-denaturation at 94 °C for 5 min, denaturation at 94 °C for 30 s, annealing at 56 °C for 30 s, extension at 72 °C for 2 min, after 30 cycles, final extension at 72 °C fo...

Embodiment 3

[0029] Example 3: Application of strain DBX666 in lignin degradation

[0030] (1) Determination of lignin degradation efficiency of strain DBX666

[0031] The strain DBX666 was inoculated into an enzyme-producing medium and cultivated to a logarithmic growth phase (OD600=0.6-0.8). Enzyme medium formulation includes: K 2 HPO 4 , 0.1g; KH 2 PO 4 , 0.4g; (NH 4 ) 2 SO 4 , 0.7125g; KNO 3 , 0.63125g; yeast extract, 0.1g; dealkalized lignin, 1-5g, adjust the pH of the medium to 7.0. The bacterial solution was inoculated into the enzyme-producing medium at a ratio of 1:100, and cultured with shaking at 20°C at 150 rpm, and the bacterial solution was taken every 24 hours to determine the lignin degradation rate of the strain. In the enzyme-producing medium, lignin, as the only carbon source, contributed to the total COD load, so the removal of COD was used to characterize the degradation rate of lignin. The determination of COD adopts the chromate method in the national stand...

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Abstract

The invention relates to a marine sediment-derived lignin-degrading bacterium and its application in degrading lignin, which is characterized in that the marine sediment-derived lignin-degrading bacterium is Rhinoceros sp. DBX666, which is preserved in Chinese microorganisms The Ordinary Microbiology Center of the Culture Collection Management Committee, the preservation number is: CGMCC No.19599, and the preservation date is April 22, 2020; compared with the prior art, the present invention has the following advantages: the strain DBX666 provided by the present invention can be It grows well in a lower medium, has a strong lignin degradation ability, and has a strong application value, realizing the maximum utilization of marine microbial resources.

Description

technical field [0001] The invention belongs to the technical field of applied microorganisms, in particular to a chitin efficient degrading bacteria suitable for marine sediment sources and its application. Background technique [0002] Lignin, one of the three most abundant biomass in nature, can be used to replace petroleum-based fuels and chemicals as a sustainable repository of organic matter second only to cellulose. At the same time, lignin is not only a by-product of wood hydrolysis industry and papermaking industry, but also a relatively refractory substance in straw and municipal solid waste, which can cause serious environmental pollution. Lignin is a polyphenolic three-dimensional network polymer aromatic compound composed of phenylpropane units connected by ether bonds and carbon-carbon bonds. Its molecular weight is large, its structure is stable, and it is difficult to degrade. It is one of the key bottlenecks in the utilization of biomass energy. . Although...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C12N9/08C12N9/02C12Q1/04C08H7/00C07G1/00C12R1/01
CPCC12N1/20C12N1/02C12N9/0065C12N9/0061C12Q1/045C08H6/00C07G1/00C12Y111/01014C12Y110/03002C12Y111/01013C12R2001/01C12N1/205Y02E50/10
Inventor 陈吉刚董冰夏朱四东杨季芳
Owner ZHEJIANG WANLI UNIV
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