Microbial preparation for urban garbage percolate treatment and application thereof
A microbial agent and microbial technology, applied in the direction of microorganism-based methods, microorganisms, biological water/sewage treatment, etc., can solve the problems of difficult landfill leachate treatment, difficult backwashing, low power consumption, etc., and achieve improved removal effect , strong adaptability, and the effect of improving the removal rate
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Embodiment 1
[0017] Preparation of liquid medium I: Take by weight 1 part of magnesium sulfate heptahydrate, 0.2 part of potassium chloride, 2.47 parts of sodium chloride, 0.3 part of sodium citrate, 1 part of yeast extract powder, 0.5 part of tryptone, add water to 100 parts , adjust the pH to be 7.0 and make liquid culture medium I;
[0018] Preparation of liquid medium II: weigh 1.2 parts of magnesium sulfate heptahydrate, 0.2 parts of potassium chloride, 2.4 parts of sodium chloride, 0.3 parts of sodium citrate, 0.2 parts of ammonium chloride, 0.5 parts of yeast extract powder, 0.15 parts of pancreatic Peptone, 0.7 part of starch, 0.5 part of glucose, add water to 100 parts, adjust pH to 7.0 to prepare liquid medium II.
Embodiment 2
[0019] Example 2 Preparation and application of microbial preparations for treating landfill leachate
[0020] (1) After activating the Erythrohalogenus bacterium Bacillus cereus, insert it into 1000 mL of the liquid culture medium I prepared in Example 1, and cultivate it at 28° C., 160 rpm, at normal pressure for 36 hours; insert the obtained bacterial liquid into 10 L of the liquid medium I prepared in Example 1 In the liquid culture medium I, at 28°C, 160rpm, normal pressure culture for 36h; then the obtained bacterial liquid was inserted into 200L liquid medium II prepared in Example 2 for large-scale production, and at 30°C, normal pressure culture for 48h to obtain Bacillus cereus suspension;
[0021] After activating the euryhalobacteria Bacillus marisflavi, insert it into 1000 mL of the liquid medium I prepared in Example 1, and cultivate it at 28° C., 160 rpm, at normal pressure for 36 h; insert the resulting bacterial liquid into 10 L of the liquid medium I prepared...
Embodiment 3
[0028] Example 3 Obtaining of transgenic euryhalohaline bacteria Bacillus cereus
[0029] Using DNAMAN software analysis, according to the amino acid sequence GenBank: WP_014597038.1 whole gene synthesis of the nucleotide sequence SL gene, the target fragment is obtained by amplification, and the target gene SL is obtained by PCR amplification (at the same time, the corresponding mutation is obtained by multiplex PCR Sites 59A / S, 82N / R, 100E / P, 123M / F, 139L / S, 147R / G, 179R / H, 191I / S, 203Q / K, 214A / R were introduced into the gene sequence, thus obtaining Different mutant genes), the PCR product was double-digested with restriction endonucleases, connected to the expression vector pSDK-1 that had also been double-digested, and the successfully verified recombinant plasmid was transferred from the large intestine by the usual method of plasmid transfer in the field. The bacilli are transformed into the euryhalobacteria bacillus cereus to obtain genetically engineered bacteria for ...
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