Chlamydomonas reinhardtii cadmium-resistant mutant, immobilized material and preparation method and application of chlamydomonas reinhardtii cadmium-resistant mutant
A rhinestone and mutant technology, applied in the field of biological wastewater treatment, can solve problems such as high cost, high energy consumption, and poor tolerance, and achieve low cost, high efficiency, and good recyclability
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Embodiment 1
[0048] Example 1 Screening of cadmium-resistant Chlamydomonas mutant
[0049]When Chlamydomonas is used to treat cadmium-containing sewage, Chlamydomonas cells often stop growing or even die due to cadmium toxicity, and the enrichment efficiency is low. In this embodiment of the present invention, a method of randomly inserting gene fragments is used to obtain mutants that are tolerant to high concentrations of cadmium. The mutants grow in a medium with a higher concentration of cadmium and can accumulate more cadmium. See the screening process figure 1 , the specific operation is as follows:
[0050] (1) Preparation of foreign insert fragments
[0051] The FSD::AphVII::RSD fragment (sequence shown in SEQ ID NO.1) in the plasmid pHK415, which contains a hygromycin antibiotic selection tag, was used as the foreign insert fragment. Adopt SpeI enzyme and ClaI double digestion plasmid pHK415, re-gel recovery FSD::AphVII::RSD fragment, its size is 1877bp ( figure 2 ).
[0052...
Embodiment 2
[0085] Embodiment 2 Chlamydomonas reinhardtii cell immobilization
[0086] Compared with the traditional immobilization technology, the present invention intends to immobilize Chlamydomonas by using sodium alginate combined with diatomaceous earth and activated carbon. On the one hand, diatomaceous earth and activated carbon can improve the porosity of the rubber balls, which is beneficial to mass transfer and promote the growth of Chlamydomonas cells in the rubber balls; The amount of exposure can further improve the ability to remove cadmium to achieve the purpose of water purification; in addition, diatomaceous earth can also increase the stability of the rubber ball and improve the strength of the rubber ball.
[0087] We explored the immobilization technology from four aspects: the mass fraction of sodium alginate, the amount of diatomite added, the amount of biochar added, and the amount of CaCl 2 Concentration is the factor, using 4 factors and 3 levels to conduct orth...
Embodiment 3
[0100] Example 3 Application of Chlamydomonas reinhardtii cell immobilization material 1
[0101] 1. Cultivate the Chlamydomonas reinhardtii mutant in TAP medium to the logarithmic phase, collect Chlamydomonas cells by centrifugation at 2500rpm for 3min, discard the supernatant, and wash with sterilized ultrapure water;
[0102] 2. Add the obtained algae liquid to the mass fraction of 2% sodium alginate, 1% diatomaceous earth, and 0.1% activated carbon, stir and mix well, and use a syringe to draw 30ml of the gel mixture and drop it in at a constant speed to a concentration of 0.1mol / L CaCl 2 In the solution, cross-link and immobilize at 20°C for 1 hour to obtain rubber balls with a diameter of about 4mm.
[0103] 3. Put the glue ball into the solution containing 250μM CdCl 2 In the wastewater, after 6 days of treatment, the content of cadmium in the water was measured by inductively coupled plasma mass spectrometry (ICP-MS), and the enrichment efficiency of cadmium was 63.41...
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