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High-temperature-resistant garden waste decomposing bacteria ST5 and application thereof

A technology of garden waste and high temperature resistance, applied in the direction of bacteria, microorganisms, microorganisms, etc., to reduce environmental pollution and accelerate the composting process

Active Publication Date: 2016-05-11
北京优生基生态科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few reports on the isolation of high temperature resistant cellulose-degrading actinomycetes from garden waste compost

Method used

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  • High-temperature-resistant garden waste decomposing bacteria ST5 and application thereof
  • High-temperature-resistant garden waste decomposing bacteria ST5 and application thereof
  • High-temperature-resistant garden waste decomposing bacteria ST5 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 ...

Embodiment 2

[0026] Embodiment 2 ST5 bacterial strain molecular biology identification

[0027] Molecular identification of Streptomyces thermovulgaris ST5 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 Gaulier No. Take out the culture medium, 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.); perform PCR amplification on the extracted bacterial DNA with universal primers 27F and 1492R; The 27F sequence was 5′-AGAGTTTGATCCTGGCTCAG-3′; the 1492R sequence was 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 length of 1418bp, the gene sequence was submitted to Ge...

Embodiment 3

[0029] Embodiment 3 Streptomyces vulgaris ST5 growth assay

[0030] Streptomyces vulgaris ( Streptomycesthermovulgaris )ST5 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 degradation bacterium which is a streptomyces thermovulgaris. The preservation number of the cellulose degradation bacterium is CGMCC No.12137. The bacterium is capable of producing a large quantity of enzymes at the high temperature of 40-70 DEG C; the cellulase of the bacterium is high in activity and has the characteristics of high temperature resistance and efficient degradation of cellulose; the bacterium can be used as a microbial agent, can be applied to a high-temperature composting system, and is suitable for composting the garden wastes.

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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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C05F11/08C12R1/465
CPCC05F11/08C12N1/20C12N1/205C12R2001/465
Inventor 周金星连鹏彭霞薇郑景明
Owner 北京优生基生态科技有限公司
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