Application of Kocuria flavus in treatment of heavy metal polluted wastewater
A technology of huangcuri and heavy metals is applied in the application field of huangcuri in the treatment of heavy metal polluted wastewater, which can solve the problems of high energy and chemical costs, and achieve the effects of environmental friendliness, low reproduction cost and huge application potential.
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Embodiment 1
[0030] Example 1 Isolation and Screening of Kocuria flavus
[0031] 1) Collect copper-contaminated soil samples with screw cap sterilized bottles;
[0032] 2) Inoculate 1g of copper-contaminated soil sample into 50ml containing 100mg / L copper solution (CuSO 4 ·5H 2 O) In the nutrient broth (NB medium), incubate for 48 hours at 30°C under shaking conditions (130 rpm);
[0033] 3) The enumeration of bacteria adopts the serial dilution full plate counting method, and the medium uses nutrient agar containing different copper concentrations (25, 50 and 100mg / L);
[0034] 4) The petri dish was incubated overnight at 30°C; subsequently, the colonies were transferred to urea agar medium (urease selection medium) to check the production of urease (because urease is a key indicator of calcite production);
[0035] 5) According to the higher urease production ability and the growth ability in copper-containing medium, finally selected Kocuria flavus; Significant results have been ach...
Embodiment 2
[0036] Example 2 Identification of Kocuria flavus
[0037] 1) The bacterial strain screened in Example 1 was cultured overnight, and the genomic DNA was extracted by alkaline lysis;
[0038] 2) The 16S rRNA gene in the genomic DNA is amplified by PCR;
[0039] 3) The 16S rRNA amplicon was gel eluted according to the manufacturer's instructions and ligated into the pTZ57R / T vector (US Enzyme Company);
[0040] 4) Sequencing was performed using the Applied Biosystems automatic sequencer; Kocuria flavus was Gram-positive and belonged to Actinomycetes and Micrococcus;
[0041] 5) The phylogenetic analysis of 16S rDNA showed that the genes of Kocuria flavus had 100% similarity with commercially available Kocuria flavus.
Embodiment 3
[0042] Example 3 Application of Kocuria flavus in the treatment of copper-polluted wastewater
[0043] 1) In a 100ml flask, add 50ml of NB medium containing 100mg / L Cu ions and 50ml of NBU medium containing 100mg / L Cu ions, respectively;
[0044] 2) Inoculate the two mediums in step 1) with Kocuria flavus respectively, such as figure 1 As shown, Kocuria flavus effectively removed 95% of copper ions (100 mg / L) in NBU medium within 120 hours; in NB medium, effectively removed 68% of copper ions.
[0045] In this example, the selected Kocuria flavus can be isolated according to the method in Example 1, or commercially available Kocuria flavus can be selected; Both Kocuria flavus and commercially available Kocuria flavus can achieve the technical effect of the present invention.
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