A kind of abnormal saccharophila Aeromonas and its application
A single cell bacteria, sugar gas technology, applied in the direction of bacteria, biological water/sewage treatment, sustainable biological treatment, etc., can solve the problems of secondary pollution, ineffective reproduction, high operating costs, etc., to achieve the effect of ensuring COD
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Embodiment 1
[0024] Example 1: Screening of oil-degrading bacteria
[0025] 1. Separation and purification: Take oily sewage samples from the sewage well of Lixiyuan canteen of Jiangnan University, add LB liquid medium to prepare a 20wt% suspension, then dilute it to an appropriate concentration and spread it on the LB solid medium plate Incubate at 37°C for 24h.
[0026] 2. Primary screening: Use an inoculation needle to select a single colony point and transfer it to the screening medium plate, culture at 37°C for 24 hours, select the strain that produces a transparent circle, insert it into 50ml LB liquid medium, and cultivate it on a shaker at 37°C and 200rpm for 8 hours. Take the cultured bacterial solution and dilute it to an appropriate concentration and spread it on an LB plate. If a single colony still produces a transparent circle, it will be used as the primary screening strain, such as figure 1 shown.
[0027] 3. Rescreening: insert the bacterium solution of the above-mention...
Embodiment 2
[0033] Embodiment 2: identification of strain
[0034] Using 16Sr DNA analysis, the oil-degrading bacteria were identified as: Aeromonas genus, Aeromonas abnormal saccharophila; scientific name Aeromonas allosaccarophila.
[0035] Extract bacterial genomic DNA, design primers 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID No.1), 1492R: 5'-GGTTACCTTGTTACGACTT-3' (SEQ ID No.2), RCR conditions: 94°C pre-denaturation for 5 minutes, Denaturation at 94°C for 30s, annealing at 55°C for 30s, extension at 72°C for 2min, 30 cycles, extension at 72°C for 10min, and incubation at 12°C.
[0036] The sequencing result is SEQ ID No.3:
[0037] AAGGTTAAGCTATCTACTTCTGGTGCAACCCACTCCCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCA
[0038] CCGCAACATTCTGATTTGCGATTACTAGCGATTCCGACTTCACGGAGTCGAGTTGCAGACTCCGATCCGGACTACGACGC
[0039] GCTTTTTGGGATTCGCTCACTATCGCTAGCTTGCAGCCCTCTGTACGCGCCATTGTAGCACGTGTGTAGCCCTGGCCGTA
[0040] AGGGCCATGATGACTTGACGTCATCCCCACCTTCCTCCGGTTTATCCCGGCAGTCTCCCTTGAGTTCCCCACCATTACGT
[00...
Embodiment 3
[0055] Embodiment 3: the preparation of bacterial agent
[0056] Aeromonas saccharophila CY01 was inoculated into liquid LB medium, cultured on a shaker at 37°C and 200 rpm for 24 hours, and then expanded in a fermenter with an inoculum size of 5% (v / v). Mix the bacterial agent and nutrient agent cultivated in the fermenter at a ratio of 1:1, pack into barrels, seal, and refrigerate at 2-8°C.
[0057] The culture medium for the expanded culture of the fermenter was prepared according to the following proportions: 5000L water, 50kg peptone, 25kg yeast powder, 50kg sodium chloride, 20kg soybean oil, pH 7.2, 121°C damp heat sterilization for 20min.
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