An engineering strain that can be used for the treatment of salt-containing organic wastewater and its application
A technology for organic wastewater treatment and engineering strains, which is applied in the field of microorganisms to achieve the effects of improving transfer efficiency, high specific surface area, and high mechanical properties
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Embodiment 1
[0043] Embodiment 1, the construction of the engineering strain that can be used for the treatment of salt-containing organic wastewater
[0044] The expression plasmid used in this example is the broad host plasmid pYYDT (the gene sequence is shown in SEQ ID NO. 6); the riboflavin expression gene cluster (controlled by the arabinose promoter, the DNA sequence is shown in SEQ ID NO. 5) ) is synthesized by General Biosystems (Anhui) Co., Ltd., and the specific construction method is as follows:
[0045] Primer 1F / 1R (the gene sequence of primer 1F is shown in SEQ ID NO.1, and the gene sequence of primer 1R is shown in SEQ ID NO.2) was used to express the gene with riboflavin synthesized by General Biosystems (Anhui) Co., Ltd. The cluster is a template, and the riboflavin expression module is obtained by amplifying the PCR instrument; using primer 2F / 2R (the gene sequence of primer 2F is shown in SEQ ID NO.3, and the gene sequence of primer 2R is shown in SEQ ID NO.4) Using the...
Embodiment 2
[0049] Example 2. Under anaerobic conditions, the comparison of the degradation rate of MO between engineered strains and wild strains
[0050] The anaerobic reaction system used in this example is a serum bottle containing 30 mL of mineral salt culture medium. After nitrogen aeration for 15 minutes, the serum bottle is sealed with a butyl rubber stopper and an aluminum cap to ensure stable anaerobic reaction in the reaction system. surroundings. After the serum bottle containing the culture medium was autoclaved at 121 °C for 20 min, MO with a final concentration of 50 mg / L (need to be filtered and sterilized through a 0.22 μm filter) was added with a syringe for use.
[0051] Strain activation: The engineering strains stored at -80°C were streaked on the LB solid medium plate, and then placed in a 30°C constant temperature incubator to cultivate until a single clone was grown. A single engineered strain on the solid plate medium was scraped with an inoculation loop and then...
Embodiment 3
[0055] Example 3. Under anaerobic conditions, the comparison of the degradation rate of roxa arsenic between engineered strains and wild strains
[0056] The anaerobic reaction system used in this example is a serum bottle containing 30 mL of mineral salt culture medium. After nitrogen aeration for 15 minutes, the serum bottle is sealed with a butyl rubber stopper and an aluminum cap to ensure stable anaerobic reaction in the reaction system. surroundings. After the serum bottle containing the culture medium was autoclaved at 121°C for 20 min, roxarsenic with a final concentration of 0.2 mM (need to be filtered and sterilized through a 0.22 μm filter) was added with a syringe for use.
[0057] Strain activation: the same as in Example 2.
[0058] The washed bacterial liquid was added into the anaerobic reaction system with a syringe, so that the bacterial concentration in the 30 mL anaerobic reaction system was OD600=0.3. The serum bottle was placed in a constant temperature...
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