Optimum growth condition of Klebsiella oxytoca strain for degrading petroleum pollutants and tolerance research method

A technology of Klebsiella and growth conditions, which is applied in the field of research on the optimal growth conditions and tolerance of Klebsiella degrading petroleum pollutants, and can solve the problems that microorganisms cannot grow and degrade petroleum, and the conditions are harsh

Inactive Publication Date: 2016-07-20
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the actual oil-polluted regional environment is very complex and the conditions are very harsh, when using microorganisms to remediate polluted waters or soils, microorganisms often cannot grow well and play a role in degrading oil.

Method used

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  • Optimum growth condition of Klebsiella oxytoca strain for degrading petroleum pollutants and tolerance research method
  • Optimum growth condition of Klebsiella oxytoca strain for degrading petroleum pollutants and tolerance research method
  • Optimum growth condition of Klebsiella oxytoca strain for degrading petroleum pollutants and tolerance research method

Examples

Experimental program
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Effect test

Embodiment 1

[0024] Determination of Klebsiella growth

[0025] Configure basic tryptone soy broth medium, peptone 20g / L, glucose 2.5g / L, sodium chloride 5g / L, dipotassium phosphate 2.5g / L, using 2mol / L sodium hydroxide and 2mol / L Hydrochloric acid adjusts the pH of tryptone soy broth medium to 7. Dispense 100 mL of Tryptone Soy Broth Medium into 250 mL Erlenmeyer flasks, seal them with parafilm, and sterilize them at 121°C for 20 minutes under high temperature and high pressure. After cooling, add 1 mL of bacterial solution, incubate on a shaker at 30°C and 220 rpm, take samples every 3 hours and measure OD 600 value. Shake the Erlenmeyer flask well before each sampling, and sample 2mL. Make three parallels for each group. In order to ensure the accuracy of the instrument during the test, control the measured value within 0.2-0.8. If the measured value is too high, do the test after appropriate dilution. OD at 3h 600 The values ​​are 0.267, 0.263, 0.234, 0.29, 0.32, 0.328 at 6h, 0.463, 0...

Embodiment 2

[0027] Determination of temperature tolerance of Klebsiella

[0028] Configure basic tryptone soy broth medium. Divide 100 mL of tryptone soy broth medium into 250 mL conical flasks, a total of 15 bottles, seal and sterilize and cool for use. Mix the Klebsiella bacteria liquid, add 1 mL of the bacteria liquid to each bottle of culture medium, and each 3 bottles form a group, respectively at 20 ℃, 25 ℃, 30 ℃, 35 ℃, 40 ℃, 220 rpm after shaking for 27 hours Sampling test. Mix and shake the flasks before sampling, and sample 2mL of each flask to test the OD of the bacterial solution 600 If the concentration is too high, the bacteria liquid should be diluted appropriately. After eliminating the measurement values ​​with obvious errors in each group, take the average value and draw the temperature tolerance diagram of Klebsiella HA1 under the condition of 20-40℃, such as figure 2 Shown. The results show that Klebsiella LF1 has the best growth at 35°C, the optimum temperature is bet...

Embodiment 3

[0030] Determination of Klebsiella pH tolerance

[0031] It is equipped with special tryptone soy broth medium for pH tolerance determination, peptone 20g / L, glucose 2.5g / L, sodium chloride 5g / L, and dipotassium hydrogen phosphate 2.5g / L. Divide 100mL of Tryptone Soy Broth Medium into 250mL Erlenmeyer Flasks, a total of 15 bottles, 3 bottles each as a group, adjust the pH with 2mol / L sodium hydroxide and 2mol / L hydrochloric acid respectively For 5, 6, 7, 8, and 9, after sealing with parafilm, sterilize at 121°C for 20 minutes under high temperature and high pressure. Mix the Klebsiella bacteria solution, add 1 mL of the bacteria solution to each bottle of culture medium, incubate at 30° C., 220 rpm for 27 hours, and then sample for testing. Mix and shake the flasks before sampling, and sample 2mL of each flask to test the OD of the bacterial solution 600 If the concentration is too high, the bacteria liquid should be diluted appropriately. After eliminating the measurement valu...

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Abstract

The invention relates to an optimum growth condition of a Klebsiella oxytoca strain for degrading petroleum pollutants and a tolerance research method. The method comprises the following steps: preparing a tryptone soya broth culture medium for use in growth situation test and tolerance condition test; performing splitting charging on the culture medium, sealing, sterilizing under high-pressure steam of 121 DEG C for 20 minutes, and cooling to the room temperature for later use; adding a strain into the tryptone soya broth culture medium for use in the growth situation test, testing the cell concentration of a culture sample through an OD600 test method, and drawing a strain growth curve; adding thalli into the tryptone soya broth culture medium for testing different tolerance conditions respectively, performing three parallel treatments for each group, culturing in a shaking table of which the speed is 220rpm, sampling after a stabilization period, testing the cell concentration through the OD600 test method, and drawing a strain tolerance curve. The Klebsiella oxytoca strain has a good oil removing effect in the temperature range of 20-40 DEG C, the acidity and alkalinity range of pH5-pH8 and the salinity range of 0.5-3 percent.

Description

Technical field [0001] The invention relates to a research method for the optimal growth conditions and tolerance of a strain of Klebsiella degrading petroleum pollutants. Background technique [0002] Petroleum pollutants refer to those petroleum and petroleum products that are uncontrollably excreted into the environment. Petroleum pollutants can have a very serious impact on the human body, animals and plants, and will pollute the environment on which humans depend, and affect water, soil and atmosphere. The environment causes varying degrees of pollution. [0003] Petroleum pollutants mainly contain one or more petroleum hydrocarbons such as alkanes, cycloalkanes, and aromatics. Petroleum pollutants may be generated in every link in the petroleum industry such as the development of oil fields and the processing of various petroleum products. . Industrial wastewater and domestic sewage constitute the main source of petroleum pollutants in environmental water, and industrial wa...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34B09C1/10A62D3/02C12R1/22C02F101/32
CPCC12N1/20A62D3/02B09C1/10C02F3/34C02F2101/32C02F2103/365
Inventor 贾晓强刘一琳李辰黄磊
Owner TIANJIN UNIV
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