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Isolated bacterial strain and application thereof

A technology of bacteria and strains, applied in the field of environmental biology, can solve the problems of insufficient degradation strains, etc., and achieve the effects of strong DMF high concentration tolerance, easy colonization, and strong DMF metabolism

Active Publication Date: 2019-01-11
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The biodegradation method is suitable for large-scale treatment of wastewater containing DMF, but there are still insufficient reports on the degradation strains, and there are still many problems to be solved urgently

Method used

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  • Isolated bacterial strain and application thereof
  • Isolated bacterial strain and application thereof
  • Isolated bacterial strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0047] The preparation method of seed liquid in the following examples is: 1) slant culture: Methylobacterium DM1 bacterial strain is inoculated on the solid slant inorganic salt (MSM) medium that contains DMF 400~2000mg / L, 25~37 ℃ cultivates 24~36 hour; 2) seed culture: the thalline with good growth on the slant in step (2) is transferred to MSM liquid medium in a sterile environment, and the DMF containing 400~2000mg / L is contained in the liquid medium , cultivated at 25-37°C for 8-24 hours to obtain seed liquid. Seed solution OD 600 0.6 to 0.8.

[0048] In the following examples, adopt HPLC method to detect the content of DMF: HPLC detection condition is: Agilent1100 high performance liquid chromatograph is equipped with XDB-C18 chromatographic column, mobile phase is 50mM NaH 2 PO 4 Aqueous solution and 0.5% acetonitrile (v / v), the flow rate is 0.5mL / min, the detection wavelength is 210nm, and the column temperature is 30°C. To quantitatively detect the DMF concentrati...

Embodiment 1

[0049] Example 1: Isolation, identification and DMF degradation characteristics of Methylobacterium DM1

[0050] 1. Strain screening and isolation

[0051] (1) Collect samples

[0052] Soil sample collection site: Shanghai Laogang Landfill.

[0053] (2) Strain isolation

[0054] The screening model is continuous subculture and isolation using DMF as the only carbon and nitrogen source and selective medium for energy.

[0055] Dissolve a total of 10 g of soil samples collected from different depths in 50 mL of distilled water, shake on a vortex shaker for 5 minutes, take 5 mL of soil solution, add it to 50 mL of inorganic salt (MSM) medium, and incubate at 30 °C, 200 rpm for 2 to 3 sky. After 3 to 4 passages, the culture solution was diluted and spread on the inorganic salt solid medium added with 2000mg / L DMF, and cultivated at 30°C for 48h to 72h. Pick a single colony into fresh MSM liquid medium, select the fastest-growing strain for streak isolation, and repeat several...

Embodiment 2

[0073] Example 2: Analysis of intermediate metabolites of DMF degraded by Methylobacterium DM1

[0074] Inoculate 15 mL of seed liquid into 1 L (containing DMF 2000 mg / L) of MSM medium, and culture at 30° C., 200 rpm. When the strain grows into the late plateau stage, collect the bacteria, resuspend with PBS buffer, and adjust the OD of the cells to 600 Adjusted to 5.0 for resting cell reaction. Add DMF (500mg / L) as the substrate to stop the cell reaction, take 10mL of the culture during the reaction from 0h to 7.5h, collect the supernatant after centrifugation (8000rpm, 10min, 4°C), and carry out the derivatization reaction, and then GC-MS analysis was performed. GC-MS can detect the intermediate metabolites dimethylamine, methylamine and ammonia.

[0075] The chemical reaction formula of the whole biocatalysis process is as follows:

[0076]

[0077] Sample derivatization method: Add 0.5 μl n-hexylamine as an internal standard to 10 mL supernatant, add 400 μl 10 mM Na...

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Abstract

The invention discloses an isolated methylbacterium sp. DM1, which can decompose N, N-Dimethyl formamide. Methylbacterium sp. DM1 can use DMF as the sole carbon and nitrogen source to grow, mineralizeit thoroughly and release ammonia nitrogen. It can tolerate and completely degrade 6800 mg / L DMF in water. Under the condition of 30 DEG C and pH 7.0, 2000 mg / L DMF, methylbacterium sp. DM1 can be degraded completely within 28 hours, and the degradation rate reaches 71.4 mg / L / h, which is one of the fastest reported strains. Methylbacterium sp. DM1 has a good application prospect in environmentalsewage treatment and environmental remediation.

Description

technical field [0001] The invention relates to the field of environmental biotechnology, in particular to an N,N-dimethylformamide degrading bacterial strain and application thereof. Background technique [0002] N,N-Dimethylformamide (DMF) is a "universal solvent" with excellent performance, which is widely used in industry, agriculture, medicine and other fields. DMF can be miscible with water and various organic solvents and has good thermal stability, but it is also a harmful volatile organic compound with poor biodegradability and difficult to effectively remove from the environment. [0003] Once DMF enters the environment, it will pose a major threat to aquatic organisms and human health. Studies have shown that DMF and its degradation products have obvious teratogenic and lethal effects on zebrafish embryos. DMF can enter the human body through the respiratory tract, digestive tract and skin, affect cell differentiation, cause chromosomal aberration, cause liver t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34B09C1/10C12R1/01C02F101/38
CPCC12N1/20C02F3/34B09C1/10B09C2101/00C02F2101/38C12R2001/01C12N1/205
Inventor 唐鸿志陆歆毓许平王伟伟陶飞
Owner SHANGHAI JIAO TONG UNIV
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