Thauera bacterium thauera sp. K11 and application thereof in degradation of phenolic compound-containing sludge wastewater
A phenolic compound, Dower's technology, used in the field of environmental protection
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Embodiment 1
[0030] Cultivation and identification of strain Thauera sp.K11
[0031] Strain cultivation and purification: Prepare solid seed culture medium; dilute the bacteria with sterile saline and spread on the above solid seed LB medium, culture in a 30°C incubator for 2-5 days, and inoculate with Single colony was picked by ring to obtain pure bacteria.
[0032] (1) Morphological characteristics
[0033] The colony of the strain Thauera sp.K11 on the LB solid medium is round, milky white, with neat edges, Gram staining is negative, and the bacteria are short rod-shaped, about 0.4-0.5 μm wide and 1.0-1.5 μm long, Such as figure 1 shown.
[0034] (2) Identification of strain 16S rRNA
[0035] Using bacterial 16S rRNA general primers 27F and 1492R as amplification primers, the 16S rRNA fragment of strain Thauera sp.K11 was amplified by PCR method, and after electrophoresis detection, it was sent to Shanghai Sangon Bioengineering Technology Service Co., Ltd. for sequencing, and the s...
Embodiment 2
[0043] Determination method of phenol content
[0044] Utilize phenolic substances in the medium of pH=10.0±0.2, in the presence of potassium ferricyanide, react with 4-aminoantipyrine to generate orange-red indoxyl antipyrine dye, and its aqueous solution is at 510nm wavelength It has the characteristics of the maximum absorption peak. Use a spectrophotometer to measure the absorbance values at different concentrations, draw a standard curve of phenol content and absorbance, and obtain a fitting calculation formula.
[0045] ①Phenol standard solution: Weigh 0.5g of colorless phenol and dissolve it in distilled water, dilute to 500ml, and make a 1g / L phenol solution;
[0046] ②Ammonia buffer solution: weigh 20g NH 4 Dissolve the white solid of Cl in 100ml of ammonia water to make an ammonia buffer solution with pH ≈ 10;
[0047] ③ 4-aminoantipyrine solution: Weigh 2g of 4-aminoantipyrine solid powder and dissolve it in distilled water, dilute to 100ml, and make a light yel...
Embodiment 3
[0056] Determination method of phenolic compound content-quantitative analysis by gas chromatography
[0057] The basis of chromatographic quantitative analysis is that the amount of the substance to be measured is proportional to its peak area (or peak height) on the chromatogram. Commonly used chromatographic analysis methods include percentage method, normalization method, internal standard method, external standard method and standard addition method. We use the internal standard method to determine the content of phenolic compounds. The advantage of the internal standard method is its high quantitative accuracy.
[0058] The phenolic compounds can be separated by using different separation times of the phenolic compounds in the gas chromatography column. The specific operation method is to use the pure substances of the components to be tested to prepare standard solutions, and then add internal standards with the same concentration to these standard solutions of equal v...
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