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Serratia marcescens strain capable of degrading chlorobenzene and application thereof

A technology of Serratia marcescens and chlorobenzene, which is applied in the field of environmental biology, can solve problems such as harsh conditions, and achieves the effects of low cost and simple components

Active Publication Date: 2020-02-18
重庆士继生态环境科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the currently reported functional strains, the degradation of chlorobenzene can only be achieved through a single form such as co-metabolism or anaerobic dechlorination, and the conditions are harsh, which limits the application of aerobic oxidation of chlorobenzene in engineering to a certain extent.

Method used

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  • Serratia marcescens strain capable of degrading chlorobenzene and application thereof
  • Serratia marcescens strain capable of degrading chlorobenzene and application thereof
  • Serratia marcescens strain capable of degrading chlorobenzene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, strain enrichment and optimization

[0033] 1) Enrichment culture: Weigh 10 g of the polluted particles, add it into a 250 ml Erlenmeyer flask filled with 100 ml of deionized water, put it into a shaker set at 30° C., and 160 r / min. After shaking and mixing thoroughly for 3 hours, take 10ml of the supernatant and inoculate it into a 250ml Erlenmeyer flask containing 100ml of LB medium, put it into a shaker set at 30°C, and cultivate it at 160r / min for 72h, then store it in a refrigerator at 4°C as enriched bacteria liquid.

[0034] 2) Isolation of pure bacteria: use cooled sterile distilled water to carry out 10-fold serial dilution of the enriched bacteria solution to make a dilution of 10 -1 、10 -2 diluent. MSM solid medium culture was carried out by pouring method. The MSM solid medium was placed in a biochemical incubator, cultured at 30°C for 3 days, and the strains with good growth (forming a single colony) were subcultured for several times, and...

Embodiment 2

[0038] Embodiment 2, strain identification

[0039]Take 1-5ml of the above-mentioned bacterial culture solution to extract genomic DNA through the bacterial genomic DNA extraction kit, and then use 6SrRNA broad-spectrum amplification primers F27: 5'-agagtttgatcatggctcag-3' (SEQ ID NO.1) and R1492: 5'-tacggttacccttgttacgactt -3' (SEQ ID NO. 2).

[0040] Use the QIAquick Genomic DNA Buffer Set to amplify the target fragment by PCR. Take 5 μl for 3% agarose gel electrophoresis, and use gel cutting to recover the target fragment for DNA sequencing. The DNA sequencing is entrusted to Sichuan Qingke Biological Company to complete. DNA sequencing was performed using Seq Forward, Seq Reverse, and Seq Internal as primers.

[0041] The identification results showed that the bacteria belonged to Gram-negative bacteria, the colony diameter was 0.5-0.8 μm, the length was 0.9-2.0 μm, the end was round, the flagella usually moved around, facultative anaerobic, most of the colonies were opa...

Embodiment 3

[0042] Embodiment 3, Serratia marcescens strain TF-1 is to p-chlorobenzene degradation effect on different substrates

[0043] Prepare benzene, toluene, formaldehyde, phenol, methanol, and ethanol with concentrations of 60mg / L, 40mg / L, 80ml / L, 40mg / L, 80ml / L, and 80ml / L respectively, and dispense them into 100ml of serum containing 50ml of MSM medium In the bottle (the concentration of chlorobenzene is 60mg / L), the above-mentioned MSM culture is sterilized under the condition of 121°C, after cooling, add 2.5ml Serratia TF-1 into the bottle with a micropipette, and cover it well Shake the rubber stopper well, put it in a shaker and set it at 30°C and 160 rpm to incubate for 156 hours, then measure the content of chlorinated olefins by gas chromatography headspace to determine the degradation effect of chlorinated olefins, and measure the concentration of bacteria liquid OD 600 nm The value determines the growth of the bacteria.

[0044] Table 1. Degradation effect of p-chlor...

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Abstract

The invention discloses Serratia marcescens strain capable of degrading chlorobenzene and application thereof. The Serratia marcescens strain has a preservation number of CCTCC NO: M 2019674, and is named Serratia marcescens TF-1. The Serratia marcescens TF-1 can degrade the chlorobenzene by taking the chlorobenzene as the sole carbon source and energy, as well as carrying out co-metabolism. In addition, the Serratia marcescens TF-1 is capable of maintaining relatively high activity in both poor-breeding polluted fields and compound polluted fields, and is suitable for application in the fields of waste water, drinking water, soil remediation and the like. Therefore, new breakthroughs can be expectedly made in the field of engineering application of biodegradation of chlorinated aromatic hydrocarbons.

Description

technical field [0001] The invention relates to the field of environmental biology technology, in particular to Serratia marcescens capable of degrading chlorobenzene, and also relates to the application of Serratia marcescens capable of degrading chlorobenzene. Background technique [0002] Chlorobenzene is a class of compounds that are artificially synthesized and relatively stable in chemical properties. It is widely used in industries such as medicine, pesticides, engineering plastics, household hygiene, material synthesis, and dye industry. However, the chlorine atom in chlorobenzene has a high electronegativity, which can strongly attract the electrons on the benzene ring, so that the benzene ring becomes an electron-phobic ring, so the chemical properties are quite stable, it is difficult to degrade, and it is highly toxic. Nervous system has inhibitory and narcotic effects, even carcinogenic; usually high chloride ion wastewater, high salinity. With the large-scale ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34B09C1/10C12R1/43C02F101/36
CPCB09C1/10C02F3/34C02F2101/36C12N1/20C12N1/205C12R2001/43
Inventor 赵天涛郭江枫邢志林王永琼曹昆刘毫
Owner 重庆士继生态环境科技有限公司
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