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High-temperature-resistant garden waste decomposition bacterium FHM1 and application thereof

A technology of garden waste and high temperature resistance, which is applied in the field of environmental biology to speed up the composting process and reduce environmental pollution

Active Publication Date: 2016-06-08
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Many cellulose-degrading bacteria are aimed at agricultural waste such as corn stalks, rice straw, and wheat straw, and there are few researches on the screening and utilization of degrading bacteria that specifically target cellulose materials from garden wastes.

Method used

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  • High-temperature-resistant garden waste decomposition bacterium FHM1 and application thereof
  • High-temperature-resistant garden waste decomposition bacterium FHM1 and application thereof
  • High-temperature-resistant garden waste decomposition bacterium FHM1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Screening of high-temperature-resistant cellulose-degrading bacterial strains

[0023] Samples were collected from composting of garden waste in an apple orchard in Changping District, Beijing at the early stage of high temperature, middle stage of high temperature, and late stage of high temperature material, and the accumulation of landscaping waste in the afforestation area of ​​Yanqing Plain in Beijing; 10 g of the above fresh samples were weighed and placed in a container containing 10 glass beads. , and filled with 90ml of sterile water in a conical flask, placed in a shaker at 30°C and 150rpm for 30min to fully disperse the samples, and left to enrich and cultivate at 50°C for 24h. Use a sterile pipette to draw 1ml of the supernatant and add it to a test tube containing 9ml of sterile water, which is 10 -1 Sample diluent, again from 10 -1 Take 1ml of the sample and add it to 9ml sterile water, which is 10 -2 sample diluent, and so on, to get 10 -3 、1...

Embodiment 2

[0026] Example 2 FHM1 strain molecular biology identification

[0027] Molecular identification of the Brevibacillus Potsdamer obtained from the screening was carried out according to the following steps: pick a single colony of the screened strain and inoculate it in liquid LB medium, culture it on a shaker at 50°C and 150 r / min, and take out the culture medium on the second day. Centrifuge at 5000r / min for 1min to take the supernatant, and extract colony DNA according to the bacterial genomic DNA extraction kit (provided by Tiangen Biochemical Technology Co., Ltd.); PCR amplification is carried out on the extracted bacterial DNA with universal primers 27F and 1492R; the sequence of 27F is 5 ′-AGAGTTTGATCCTGGCTCAG-3′; 1492R sequence is 5′-AAGGAGGTGATCCAGCCGCA-3′; the PCR product was sequenced, and the sequencing results were analyzed by BLAST in the NCBI database and compared for homology.

[0028] The 16S rDNA gene sequence of Brevibacillus Potsdam (see figure 2 ) with a l...

Embodiment 3

[0029] Embodiment 3 Brevibacillus Potsdam growth assay

[0030] Brevibacillus Potsdam strain FHM1 was inoculated into CMC liquid medium (CMC-Na15.0g, NH 4 NO 3 1.0g, yeast extract 1.0g, MgSO 4 0.5g, KH 2 PO 4 1.0g, distilled water 1000mL), take the culture solution every 2h, continuously sample for 48h, measure the OD600 value of each period, take the culture time as the abscissa, and the OD600 value of each sampling point as the ordinate, draw the growth curve of the bacteria , the growth assay of the bacteria. It can be seen from the measurement results that 0-8h is the lag phase, 9-16h is the logarithmic growth phase, 16-24h is the stable phase, and >26h is the decline phase. The strains in the logarithmic phase grow rapidly and have strong vitality. Therefore, in the subsequent fermentation enzyme production experiments, the 12-hour fermentation liquid should be used as the seed liquid for inoculation.

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Abstract

The invention discloses a high-temperature-resistant cellulose decomposition bacterium Brevibacillus borstelensis, collected under CGMCC No. 12138. This bacterium can produce massive enzymes at a high temperature of 40-70 DEG C, has high cellulose activity, features high temperature resistance and efficient cellulose degradation, can be used as a microbial agent for application in high-temperature composting systems and is applicable to garden waste composting.

Description

technical field [0001] The invention belongs to the field of environmental biotechnology, and in particular relates to a high-efficiency cellulose-degrading heat-resistant bacterium screened from garden waste composting and its application in garden waste high-temperature composting. Background technique [0002] Garden waste refers to the dead branches, fallen leaves, grass clippings, residual flowers, tree and shrub pruning and other plant residues produced by the natural fall of garden plants or artificial pruning. The main components are refractory cellulose and hemicellulose. In recent years, my country's garden waste has been increasing at an annual rate of 8%-10%, which has brought great resistance to the process of urban greening. At present, the main methods of dealing with garden waste in my country are incineration and filling. These two processing methods are not only wasteful The use of renewable resources has also caused serious environmental pollution. Therefo...

Claims

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Application Information

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IPC IPC(8): C12N1/20C05F15/00C05F17/00C12R1/13
CPCC05F3/00C05F17/00C12N1/205C12R2001/13C05F11/00C05F11/08Y02W30/40
Inventor 彭霞薇冯红梅周金星郑景明
Owner BEIJING FORESTRY UNIVERSITY
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