A strain of Brevibacillus thermophilic producing amylase and its application
A Bacillus and Bacillus licheniformis technology, applied in the field of environmental biology, can solve the problems of perishable deterioration, high moisture content of kitchen waste, environmental pollution, etc., and achieve the effects of good starch degradation ability, shortening heating time, and increasing maximum temperature.
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Embodiment 1
[0048] The screening of embodiment 1 thermophilic brevibacillus B205
[0049] (1) Mix food waste, residual sludge from food factory sewage treatment, and corn straw at a ratio of 0.85:0.85:0.8, and inoculate 5% (w / w) of rotten rice straw in a certain district of Wuxi City, Jiangsu Province. The moisture content is 55%, the C / N ratio is 25, and the compost is composted after mixing evenly. The temperature of the compost is measured at 8:00 in the morning and 6:00 in the afternoon. As the composting time prolongs, the temperature of the compost body begins to drop. When the temperature is lower than 40°C, stop turning the compost and take samples;
[0050] (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 and 10 -6 100 μL of the above three dilutions were drawn and spread evenly on the nutrient agar medium, Gao’s ...
Embodiment 2
[0052] The identification of embodiment 2 brevibacillus thermophiles B205
[0053] After culturing overnight at 45°C on a nutrient agar medium plate, white, oval, smooth-surfaced, irregular-edged colonies are formed, which are Gram-positive, rod-shaped, spore-forming, and produce a large amount of foreign substances when observed under an optical microscope. polysaccharides.
[0054] 16SrDNA sequencing was used for taxonomic molecular identification, and the sequencing results (1469bp) were compared by Blast in NCBI, and it was found that the 16SrDNA base sequence had the highest similarity with Brevibacillus thermoruberstrain (99%).
[0055] Based on the above morphological characteristics, physiological and biochemical characteristics and genetic characteristics, the amylase-producing strain can be identified as Brevibacillus thermoruber strain (Brevibacillus thermoruber strain), named it as Brevibacillus thermoruber strain B205, and was released on July 2018. It was deposi...
Embodiment 3
[0058] Embodiment 3 Thermophilic Brevibacillus B205 strain activation and cultivation
[0059] Brevibacillus thermophilic B205CCTCC No.M 2018472 was inoculated onto a nutrient agar medium plate and cultured at 45°C for 22h. The composition of the nutrient agar medium is 15.0 g of glucose, 5.0 g of yeast extract, and 0.2 g of calcium chloride (with two crystal waters).
[0060] The activated strains were inoculated into 250 mL shake flasks filled with 100 mL of liquid medium, and cultured with shaking at 500 r / min for 22 h at 45 °C. The shake flask medium consists of 15.0 g of glucose, 5.0 g of yeast extract, 0.2 g of calcium chloride (with two crystal waters), 1000 mL of water, and a pH of 7.0.
[0061] 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 0.8-1.5vvm, the dissolved oxygen is greater than 30%, and the temperature is 30°C-60°C Fermentation culture 20...
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