Bacterial strain for repairing chromium slag contaminated site
A site remediation and strain technology, applied in the field of chromium-reducing functional bacteria
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Embodiment 1
[0011] Example 1: Separation, purification and domestication of the chromium reducing bacteria of the present invention
[0012] Collect the soil under the chromium residue storage site of Hunan Ferroalloy Plant, weigh 10g of the chromium residue soil into a 150mL sterilized Erlenmeyer flask, add 10mL sterilized liquid medium (5g / L sodium lactate, 5g / L yeast extract and 2g / L Sodium chloride, the rest is water, use 5mol / L NaOH to adjust pH=9.5~9.8), culture in 30℃ incubator for 4 days. Then take 1mL of mud and dilute it to 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 , Using a pipette from 10 -6 , 10 -7 And 10 -8 Take 0.1 mL of the diluent and place it with 250mg / L Cr(VI) (in K 2 Cr 2 O 7 The solution is used as the solid medium for the chromium source), and the solid medium is composed of 5 g / L sodium lactate, 5 g / L yeast extract, 2 g / L sodium chloride, and 15 g / L agar. Fully spread with a triangular rod, and place the plate upside down in a 30°C incubator for 4 days. Use a ste...
Embodiment 2
[0015] Example 2: Bacteriological characteristics and biological identification of strains
[0016] Take a little drop of the bacteria liquid on the glass slide, allow the bacteria to attach for 15 minutes, put it in a 2.5% glutaraldehyde solution and fix for 1 hour, dehydration with an alcohol gradient of 30% to 70%, replace with isoamyl acetate for 30 minutes, and dry at the critical point , Ion sputtering, scanning electron microscope observation and taking pictures, see attached figure 1 .
[0017] 1. Morphological characteristics
[0018] (1) Colony morphology characteristics: the liquid medium with chromium-reducing bacteria is inoculated into a solid medium containing 250mg / L of hexavalent chromium by the streaking plate method, and the plate is placed upside down in a 30°C constant temperature incubator. After 3 to 4 days, observe the morphology of the colony. On the plate with Cr(VI), the colony is blue-gray, round, with neat edges, and a protrusion in the middle. figure 1 ...
Embodiment 3
[0030] Example 3: Reduction of Cr(VI) by chromium reducing bacteria
[0031] Bacterial reduction Cr(VI) conditions: medium components: 5g / L sodium lactate, 5g / L yeast extract, 2g / L sodium chloride; adjust pH=9.5~9.8 with 5mol / L NaOH; temperature 30℃.
[0032] Use an inoculating loop to pick the purified colony into an Erlenmeyer flask filled with 100 mL sterile culture medium. Incubate at 30°C and a 150 rpm shaker for 24 hours. Then, use a sterile pipette to transfer 10 mL of the overnight growth , Inoculate into 100mL sterile culture medium with hexavalent chromium concentrations of 25, 50, 100, 150, 200, 300, 400 and 500 mg / L (with K 2 Cr 2 O 7 Solution as the source), incubate at 30°C, 150rpm, and shaker for 72h. During this period, at 0, 6, 12, 24, 36, 48, 60 and 72h after incubation, sample with a sterile syringe, centrifuge, and supernatant Carry out the determination of residual hexavalent chromium, the results of the determination of Cr(VI) in each period are as attached F...
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