A heat-resistant garden waste decomposing bacterium st5 and its application
A garden waste, high temperature-resistant technology, applied in the direction of bacteria, microorganisms, microorganisms, etc., to reduce environmental pollution and accelerate the process of composting.
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Embodiment 1
[0022] Example 1 Screening of high-temperature-resistant cellulose-degrading 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 90 ml of sterile water in a conical flask, placed in a shaker at 30°C and 150 rpm for 30 minutes to fully disperse the samples, and left to enrich at 50°C for 24 hours. Use a sterile pipette to draw 1 ml of the supernatant and add it to a test tube containing 9 ml 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 、10 -4 、10 ...
Embodiment 2
[0026] Example 2 ST5 strain molecular biology identification
[0027] Molecular identification of Streptomyces thermovulgaris ST5 obtained through screening was carried out in accordance with the following steps: pick a single colony of the screened strain and inoculate it in liquid Gowell 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 is 5′-AGA GTT TGA TCC TGG CTC AG-3′; the 1492R sequence is 5′-AAG GAG GTG ATC CAG CCGCA-3′; the PCR product is sequenced, and the sequencing results are analyzed by BLAST in the NCBI database , and perform homology comparisons.
[0028] The 16S rDNA gene sequence of Streptomyces thermovulgaris (see figure 2 ) was 1418bp in length, and the gene sequence was su...
Embodiment 3
[0029] Embodiment 3 Streptomyces vulgaris ST5 growth assay
[0030] Streptomyces vulgaris ( Streptomycesthermovulgaris ) ST5 was inoculated into CMC liquid medium (CMC-Na 15.0g, 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 broth should be used as the seed solution for inoculation.
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