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Method for separating and purifying Ochrobactrum sp. bacterium used for reducing Cr6+

A technology of separation and purification, mgso4·7h2o0.2g, applied in the field of microbial cultivation, can solve the problems of difficult separation and cultivation of microorganisms, achieve the effects of reducing workload and cost, high colony formation rate, and simple cultivation components

Inactive Publication Date: 2008-07-09
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to solve the present microbial treatment alkaline Cr-containing 6+ For the problem that microorganisms are difficult to separate and cultivate in wastewater, a single-layer plate and improved solid medium for separation and purification are provided to obtain Cr-reducing 6+ Ochrobactrum sp. bacteria pure culture method, it can be applied to the treatment of alkaline chromium-containing wastewater, chromium slag leachate, especially chromium slag, etc., and the method is simple and the colony formation rate is high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0008] Embodiment 1: Separation and reduction of Cr 6+ Ochrobactrum sp. strain Cr-1.

[0009] Add 3.0g of the chromium residue sample collected in the chromium salt factory into 100mL of sterilized LB medium, enrich and cultivate for one generation at 30°C and 160r / min, take 2ml of the supernatant and inoculate it into a new selective liquid Culture medium Cr-G (100ml), the composition of selective liquid medium Cr-G medium is: peptone 10.0g / L; yeast extract powder 5.0g / L; NaCl 5.0g / L; MgSO 4 ·7H 2 O 0.2g / L; K 2 HPO 4 0.05g / L; K 2 Cr 2 o 7 0.8g / L. The pH was adjusted to 7.5 with NaOH solution, the culture temperature was 30° C., and the subculture was carried out for 3 times, and the inoculum size for each time was 2 ml (2%). Finally, an appropriate amount of the culture in the logarithmic growth phase was diluted 20 times, and an appropriate amount of the diluted solution was dipped with an inoculation loop and streaked on a solid plate. The solid plate medium form...

Embodiment 2

[0011] Embodiment 2: Separation and reduction of CR 6+ Ochrobactrum sp. strain Cr-2

[0012] From enrichment culture to selective liquid medium Cr-G subculture, the steps are the same as in Example 1, K 2 Cr 2 o 7 It is 0.9g / L, medium pH=9.0, and its culture temperature is still 30 ℃, subcultured 2 times, will be in the logarithmic growth phase culture fluid and dilute 60 times at last, carry out solid medium plate with embodiment 1 step coating separation, K 2 Cr 2 o 7 Still 0.4g, the concentration of agar powder is 1.1%, the pH is adjusted to 9.0, and the culture temperature is set to 30°C. Parts A and B were steam sterilized at 121-125°C for 20 minutes, respectively. Spread the diluted bacterial solution evenly with a glass rod, and streak culture for about 2 days to form more than 60 gray, raised, and round colonies on the surface of the medium, but the diameter of the colonies this time is smaller than that of Example 1.

[0013] The obtained single colonies were ...

Embodiment 3

[0014] Embodiment 3: Separation and reduction of Cr 6+ Ochrobactrum sp. strain Cr-3

[0015] From enrichment culture to selective liquid medium Cr-G subculture, the steps are the same as in Example 1, but the sample collected is about 0.5 meters from the edge of the chromium slag heap, 4.0g of soil samples 10cm deep, and the selective liquid medium Cr -K in G 2 Cr 2 o 7 Adjust to 1.0g / L, culture medium pH=9.0, its culture temperature is 30 ℃, subculture 3 times, inoculum size is 2mL (2%), will be in the logarithmic growth phase culture fluid diluted 80 times at last, with implementation Carry out streak separation on solid medium plate in the way of Example 1, K 2 Cr 2 o 7 The setting is 0.5g, the concentration of agar powder is 1.1%, the pH is adjusted to 9.0, and the culture temperature is set to 37°C. Parts A and B were steam sterilized at 121-125°C for 20 minutes, and streak cultured for about 2-3 days to form dozens of gray round single colonies with a convex middl...

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Abstract

The invention discloses a method for separation and purification of Ochrobactrum sp. with deoxidizing Cr 6+, which solves the problem of that prior microorganism is difficult to isolated culture when microorganism disposes alkali waste water with Cr 6+. Picking alkali waste soil sample with Cr 6+ which is inoculated on organic LB medium to process enriched culture generation, adopting selective liquid medium Cr-G which is improved by inventor to process passage culture, and then adopting single-layer flat-plate and improved solid medium to process streak culture. The invention is capable of realizing separation and purification of Ochrobactrum sp. with deoxidizing Cr 6+ in alkality environment, adopting single-layer solid medium and optimize culture condition, wherein culture component is simple and formation rate of bacterial colony is higher.

Description

technical field [0001] The present invention relates to a kind of microbial cultivation method, especially for Cr 6+ Method for the isolation and purification of pure cultures of reduced Ochrobactrum sp. bacteria. Background technique [0002] Wastewater discharged from mines, smelting, inorganic salt and chromium salt factories, etc. has high content of hexavalent chromium and high toxicity. In recent years, the use of microorganisms to treat Cr-containing 6+ The method of waste water is very attractive, and the biological method has the characteristics of simplicity, cheapness and good effect. Various microorganisms are constantly being developed and utilized, such as: Pseudomonas ambiguus, Pseudomonas fluorescens, Streptococcus lactis, etc. Although these strains have certain resistance to chromium (VI), they cannot change the chromium (VI) content. The valence state of chromium (VI) cannot be reduced to reduce its toxicity, and the purpose of treating chromium (VI) wa...

Claims

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Application Information

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IPC IPC(8): C12N1/02C12N1/20C12R1/01
Inventor 贺治国高凤玲胡岳华何超
Owner CENT SOUTH UNIV
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