A heat-resistant garden waste decomposing bacterium st1 and its application
A technology of garden waste and high temperature resistance, applied in the direction of bacteria, microorganisms, microorganisms, etc., to achieve the effect of reducing environmental pollution and accelerating the process of composting and decomposing
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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 ST1 strain molecular biology identification
[0027] Molecular identification of the Streptomyces hyperthermophilus 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 Gowell No. 1 medium, culture it on a shaking table at 30°C and 120r / min, and take it out on the 2nd day The culture solution was centrifuged at 5000r / min for 1min to take the supernatant, and the colony DNA was extracted according to the bacterial genomic DNA extraction kit (provided by Tiangen Biochemical Technology Co., Ltd.); the general primers 27F and 1492R were used for PCR amplification of the extracted bacterial DNA; 27F The sequence is 5′-AGAGTT TGA TCC TGG CTC AG-3′; the sequence of 1492R is 5′-AAG GAG GTG ATC CAG CCG CA-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 seque...
Embodiment 3
[0029] Example 3 Determination of the growth of Streptomyces violaceum
[0030] Streptomyces hyperthermophilus ST1 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 decay 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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