Rhodococcus JZX-01 used for degrading petroleum pollutants, and culturing method thereof

A JZX-01, culture method technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve problems such as low solubility, low utilization rate of petroleum hydrocarbons, and decreased ability to biodegrade petroleum, and achieve degradation High efficiency, increase mutual emulsification ability, and reduce surface tension

Inactive Publication Date: 2013-01-02
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Due to the extremely low solubility of petroleum hydrocarbons in water, the utilization rate

Method used

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  • Rhodococcus JZX-01 used for degrading petroleum pollutants, and culturing method thereof
  • Rhodococcus JZX-01 used for degrading petroleum pollutants, and culturing method thereof
  • Rhodococcus JZX-01 used for degrading petroleum pollutants, and culturing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Screening of Rhodococcus JZX-01

[0048] The polluted soil was retrieved from the area seriously polluted by oil in the Bohai Rim region. During the process of collecting soil samples, soil samples were taken at different depths and locations. The retrieved soil sample is used to add the inducer to the culture medium or culture medium, and add the soil to the culture medium or culture medium, and cultivate it in a shaker at a temperature of 10-40°C and a speed of 100 -300rpm. After culturing for 1-4 days, transfer to fresh medium or culture solution for screening. Wherein, the medium components are ammonium sulfate 5g / L, NaCl 0.5g / L, trace element mixed solution 10mL / L, dipotassium hydrogen phosphate 3g / L, potassium dihydrogen phosphate 3g / L, add distilled water to 1L . The added inducer is 5mL / L, the culture temperature is 30° C., the pH value is 7.0, and the inducer is diesel oil. The above-mentioned medium or culture solution were all sterilized at 121° C. for 30...

Embodiment 2

[0050] Purification of Rhodococcus JZX-01

[0051] Dilute the obtained bacterial solution and spread it on a solid nutrient medium. The components of the solid nutrient medium are: soluble starch 2g / L, ammonium sulfate 5g / L, NaCl 0.5g / L, trace element solution 10mL / L, agar 20g / L, monovalent metal ion of phosphate or its hydrate 3g / L, dipotassium hydrogen phosphate 3g / L, potassium dihydrogen phosphate 3g / L, add distilled water to 1L. Pick a single colony for preservation. After obtaining a single colony, verify the degradation ability of each strain on diesel oil, and one of the strains with the best degradation ability is named JZX-01.

Embodiment 3

[0053] The Effect of Carbon Source on the Growth of Rhodococcus JZX-01

[0054] The obtained Rhodococcus JZX-01 was expanded in a liquid nutrient medium. The carbon sources of the liquid nutrient medium were soluble starch 2g / L, glycerol 2g / L, glucose 2g / L, and the nitrogen source was ammonium sulfate 5g / L. , NaCl 0.5g / L, trace element solution 10mL / L, dipotassium hydrogen phosphate 3g / L, potassium dihydrogen phosphate 3g / L add distilled water to 1L. The effects of the above three carbon sources on the growth of the bacteria were investigated separately, and it was found that the growth of JZX-01 was the best in the medium supplemented with 2 g / L soluble starch.

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Abstract

The invention relates to a rhodococcus JZX-01 used for degrading petroleum pollutants, and a culturing method thereof. The identification of the rhodococcus is achieved through 16S rDNA sequencing and comparison on NCBI. An adopted 16S rDNA upstream primer is a universal primer 27F with a specific sequence of 5'-AGA GTT TGA TCC TGGCTCAG-3'. A downstream universal primer is 1492R with a specific sequence of 5'-GGT TAC CTT GTT ACG ACTT-3'. According to the invention, an inducing agent is added into a culture medium or culture solution, and soil is added into the culture solution or culture medium; culturing is carried out in a shaking table under a culturing temperature of 10-40 DED C and a culturing rotation speed of 100-300rpm; when culturing is carried out for 1-4days, a product is transferred into a fresh culture medium or culture solution, and screening is carried out. The rhodococcus strain is collected by China General Microbiological Culture Collection Center, and has a collection register number of CGMCC No. 5958. The collection date is April 9th, 2012. With a surfactant-producing capacity of rhodococcus JZX-01, petroleum degradation efficiency is further enhanced. The rhodococcus JZX-01 can be used in biological treatments of petroleum pollutants.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a cultivation method and application of Rhodococcus sp. JZX-01, CGMCC No.5958, (Rhodococcus sp. role in the process. Background technique [0002] Petroleum and its derivatives play a very important role in daily life and industrial production. However, the discharge of oil in the process of mining, smelting, storage, transportation and use has caused huge damage to the environment, and the frequent oil spills around the world have exacerbated this harm. Most of the waters on the earth, especially the oceans, have been polluted by oil all year round, which has aroused great concern all over the world. [0003] Different components in petroleum have different effects on humans, animals and plants. Saturated hydrocarbons with low boiling points can cause paralysis, coma and, in high concentrations, even death. Hydrocarbons with high boiling points are eas...

Claims

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Application Information

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IPC IPC(8): C12N1/20A62D3/02C12R1/01A62D101/20
Inventor 贾晓强周征西闻建平
Owner TIANJIN UNIV
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