Raoultella sp. and application thereof in pyrene-Cr(VI) composite pollution co-detoxication
A compound pollution, -cr technology, applied in the field of environmental microorganisms, can solve the problems of high energy consumption, high cost, secondary pollution, etc.
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Embodiment 1
[0114] Embodiment 1: Isolation and identification of pyrene-Cr (VI) co-detoxification bacteria
[0115] (1) Primary screening
[0116] Samples were collected from the soil around the oil tank and the soil around the coal pile near Longwan Wharf, Oujiangkou, Wenzhou City, Zhejiang Province. The depth of soil sample collection was 0-10 cm, and the sampling volume at each point was greater than 100 g. Weigh 10g of each sample and place it in a Erlenmeyer flask filled with 90ml of sterile water, shake it well, take 5ml and inoculate it into a Erlenmeyer flask with 50ml of inorganic salt medium, and enrich and cultivate at 30°C and 150r / min . Basic composition of inorganic salt medium: (NH4)2SO4 1g, MgSO4 7H2O 0.2g, CaCL2 0.2g, NaH2PO4 0.5g, K2HPO4 0.5g, NaCL 0.2g, Fe2(SO4)3 7H2O 0.01g, distilled water 1000ml, pH 7.5 Add Cr (Ⅵ) (K2Cr2O7) 10mg / L, pyrene (n-hexane solution) 50mg / L in the basic medium. After the bacteria grow out (cultivate for about a week), transfer to a new inor...
Embodiment 2
[0135] Example 2: Conditions for co-detoxification of pyrene-Cr(Ⅵ) by Raoultellasp.KX255631
[0136] Strain Raoultella sp.KX255631 (1.5ml frozen tube thawed bacteria solution) was inoculated into a 250ml Erlenmeyer flask filled with 50ml LB medium, cultured at 30°C and 150r / min for 24h to activate the strain; The volume fraction of five was transferred to a 500ml Erlenmeyer flask containing 100ml LB medium, cultured at 30°C and 150r / min for 24h, centrifuged at 8000r / min for 10min, washed the bacteria twice with sterile water, and then washed with sterile water Adjusted to OD with sterile water 600 = 0.999-1.000 bacterial suspension; put into a 250ml Erlenmeyer flask equipped with 50ml inorganic salt medium according to the volume fraction of 10%, vibrate and cultivate at a certain temperature and rotating speed for one week, and measure the starting and ending inorganic salt culture respectively Cr (VI) and pyrene concentration in the base (basic composition is the same as in...
Embodiment 3
[0156] Embodiment 3: Raoultellasp.KX255631 remediation soil pyrene-Cr (VI) composite pollution test
[0157] The pyrene-Cr(VI) co-detoxification effect of Raoultella sp.KX255631 on artificial pyrene-Cr(VI) co-contaminated soil was studied by using higher plant toxicology test method. The specific method is: collect soil at a depth of 0-20 cm from the nursery of Wenzhou University without pyrene and Cr(VI) pollution, remove stones and plant residues, pass through a 50-mesh sieve, air-dry and mix evenly, and measure the physical and chemical properties of the soil (Table 5 ); after the naturally air-dried soil was sterilized at 121°C for 20 minutes, 50 g was weighed in a sterilized petri dish, a total of 27 dishes, divided into three groups: group A added sterile deionized water, group B added pyrene 50 mg / Kg soil, Cr(VI)) 40mg / Kg soil and sterile deionized water, group C added pyrene 50mg / Kg soil, Cr(VI)) 40mg / Kg soil, Raoultella sp.KX255631 bacterial suspension and sterile de...
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