High-temperature-resistant garden waste decomposing bacterium ST4 strain and application thereof

A technology of garden waste and high temperature resistance, applied in the field of environmental biology, to achieve the effect of accelerating the process of composting and reducing environmental pollution

Active Publication Date: 2016-06-15
北京优生基生态科技有限公司
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  • Abstract
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  • 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 bacterium ST4 strain and application thereof
  • High-temperature-resistant garden waste decomposing bacterium ST4 strain and application thereof
  • High-temperature-resistant garden waste decomposing bacterium ST4 strain 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] Embodiment 2 ST4 bacterial strain molecular biology identification

[0027] Molecular identification of the screened Streptomyces albicans was carried out in accordance with the following steps: pick a single colony of the screened strain and inoculate it in liquid Gowell No. 1 medium, culture it on a shaker at 30°C and 120 r / min, and take it out for culture on the 2nd day solution, centrifuged at 5000r / min for 1min to take the supernatant, and extracted colony DNA according to the bacterial genomic DNA extraction kit (provided by Tiangen Biochemical Technology Co., Ltd.); general primers 27F and 1492R were used for PCR amplification of the extracted bacterial DNA; 27F sequence It is 5′-AGAGTTTGATCCTGGCTCAG-3′; the 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 ...

Embodiment 3

[0029] Embodiment 3 Streptomyces albicans growth assay

[0030] Inoculate Streptomyces albicans ST4 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 12h, continuously sample until 120h, 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 0-24h is the lag phase, 24-72h is the logarithmic growth phase, 72-96h is the stable phase, and >96h is the decline phase. The strains in the logarithmic phase grow rapidly and have strong vitality. Therefore, in the subsequent fermentation enzyme production experiment, the 48h fermented liquid should be used as the seed liquid for inoculation.

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Abstract

The invention discloses a high-temperature-resistant cellulose degrading bacterium strain Streptomyces albidoflavus of which the collection number is CGMCC No.12136. The strain can produce abundant enzymes at the high temperature of 40-70 DEG C, has the characteristics of high cellulase activity, high temperature resistance and high cellulose degradation efficiency, can be used as a microbial inoculant in a high-temperature composting system, and is suitable for 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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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C05F11/00C05F11/08C05F15/00C12R1/465
CPCC05F3/00C05F11/00C05F11/08C12N1/205C12R2001/465C05F9/04
Inventor 周金星彭霞薇冯红梅连鹏
Owner 北京优生基生态科技有限公司
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