Bacillus thermoamylovorans strain and application thereof
A technology of Bacillus amyloliquefaciens and Bacillus spp. is applied in the field of environmental biology, which can solve the problems of long decomposing time of compost and difficult degradation of garden waste, and achieve the effects of prolonging the duration of high temperature, shortening the composting cycle and shortening the heating time.
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Embodiment 1
[0047] The screening of embodiment 1 thermophagocytic bacillus amylophagous B202
[0048] (1) Mix the excess sludge from municipal sewage treatment plants, excess sludge from food factory sewage treatment, and corn straw at a ratio of 0.75:0.75:1, and inoculate 5% (w / w) of rotten rice straw in a certain district of Wuxi City, Jiangsu Province, The moisture content of the mixed material is 55%, and the C / N ratio is 25. Compost the compost after mixing evenly. Measure the temperature of the compost at 8:00 am and 6:00 pm every day. When the temperature is higher than 55°C, turn the compost once a day , as the composting time prolongs, the temperature of the compost begins to drop, and when the temperature is lower than 40°C, stop turning and take samples;
[0049] (2) Weigh 10 g of compost samples, put them into a Erlenmeyer flask with 90 mL of sterilized water, and vibrate on a shaking table at 120 r / min for 1 h to obtain 10 -1 dilutions, and sequentially made 10 -4 、10 -5 a...
Embodiment 2
[0051] The identification of embodiment 2 Bacillus amylophagous B202
[0052] After culturing overnight at 45°C on a nutrient agar medium plate, the formed colony is cream-colored, with irregular edges, slightly rough, 1-4mm in diameter, and is positive for Gram staining by optical microscopy, single or 2-4 The cells are arranged in a chain, rod-shaped, spore-forming, and can use carbohydrates to produce acid.
[0053] 16SrDNA sequencing was used for taxonomic molecular identification, and the sequencing results (1493bp) were compared by Blast in NCBI, and it was found that the 16SrDNA base sequence had the highest similarity with Bacillus thermoamylovorans strain, which was 99%.
[0054] Based on the above morphological characteristics, physiological and biochemical characteristics and genetic characteristics, the laccase-producing strain can be identified as Bacillus thermoamylovorans strain, named Bacillus thermoamylovorans strain B202, and was released on July 13, 2018. I...
Embodiment 3
[0057] Embodiment 3 Bacillus amylophagous B202 strain activation and cultivation
[0058] Bacillus amylophagous B202 was inoculated on a nutrient agar medium plate and cultured at 45°C for 22 hours. The composition of the nutrient agar medium is 3.0 g of beef extract, 10.0 g of peptone, and 5.0 g of sodium chloride.
[0059] The activated strains were inoculated into a 250 mL shake flask filled with 100 mL of liquid medium, and cultured with shaking at 160 r / min for 22 hours at 45°C. The shake flask culture medium consists of 3.0 g of beef extract, 10.0 g of peptone, 5.0 g of sodium chloride, and a pH of 7.3.
[0060] Inoculate the shake flask culture solution into a 3L fermenter according to the volume ratio of 3%-10%, the stirring speed is 200-600r / min, the ventilation volume is 0.5-1.0vvm, the dissolved oxygen is greater than 10%, and the fermentation is carried out at 30-60°C 20-30h. The medium composition of the fermenter is 20.0 g of peptone, 30 g of glucose, 2.0 g of...
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