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An isolated bacterial strain and its application

A technology of bacteria and strains, applied in the field of environmental biology, can solve problems such as insufficient degradation strains

Active Publication Date: 2022-02-22
SHANGHAI JIAOTONG 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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  • An isolated bacterial strain and its application
  • An isolated bacterial strain and its application
  • An isolated bacterial strain and its application

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0047] The preparation method of seed fluid in the following examples is: 1) Slamp Culture: The methyl bacillus DM1 strain is inoculated to a solid beveled inorganic salt (MSM) medium containing DMF 400 to 2000 mg / L, 25 ~ 37 ° C culture 24 ~ 36 Hours; 2) Seed culture: Turn a good bacterium in the inclined surface in step (2), transferred to the MSM liquid medium in a sterile environment, the liquid medium containing 400 to 2000 mg / L DMF The seed fluid was obtained at 25 to 37 ° C for 8 to 24 hours. Seed liquid OD 600 It is 0.6 ~ 0.8.

[0048] In the following examples, the content of DMF is detected by HPLC method: The HPLC detection condition is: Agilent1100 high-performance liquid chromatograph with XDB-C18 column, mobile phase 50mm Nah 2 PO 4 The aqueous solution and 0.5% acetonitrile (V / V) were 0.5 ml / min, the detection wavelength was 210 nm, and the column temperature was 30 ° C. The DMF concentration was quantified with HPLC, and the DMF standard of different concent...

Embodiment 1

[0049] Example 1: Isolation, Identification and DMF Degradation Characteristics of Methyl Bacillus DM1

[0050] 1, strain screening and separation

[0051] (1) Collecting samples

[0052] Soil sample collection location: Shanghai Old Harbor Waste Fill.

[0053] (2) Separation of strains

[0054] The screening model is continuously passing and separated by selective medium with DMF as a unique carbon nitrogen source and energy.

[0055] The soil samples collected from different depths were dissolved in 50 ml of distilled water, oscillated on the vortex oscillator for 5 min, and 5 ml of soil solution was added to 50 ml of inorganic salt (MSM) medium, at 30 ° C, 200 rpm culture 2 ~ 3 sky. After 3 to 4 times, the culture solution was diluted to add 2000 mg / l DMF in inorganic salt solid medium, and culture was cultured from 48 h to 72 h. In the fresh MSM liquid medium, the simple MSM liquid medium is selected, and the fastest strain is selected for the line separation, repeats multip...

Embodiment 2

[0073] Example 2: Analysis of Intermediate Metabolism of DMF DM1 DMF DMF

[0074] 15 ml of seed liquids were inoculated into 1 l (containing DMF 2000 mg / L) MSM medium, at 30 ° C, 200 rpm culture. When the strain grows into the platform period, the bacteria is collected, resuspended with PBS buffer, and the OD of the cells 600 Adjusts to 5.0 to carry out a stop cell reaction. DMF (500 mg / L) was added as a substrate to carry out cellular reactions, and 10 ml of culture was taken from 0H to 7.5 hours, and the supernatant after centrifugation (8000 rpm, 10 min, 4 ° C) was collected, and the derivatization reaction was performed. Perform GC-MS analysis. GC-MS detects intermediate metabolite dimethylamine, methylamine and ammonia.

[0075] The chemical reaction formula of the entire bio catalytic process is as follows:

[0076]

[0077] Sample Derivatization Method: 0.5 μl of n-hexamethylamine was added to 10 ml of supernatant, add 400 μl of 10 mMNAOH, 100 μl of benzenesulfonyl ch...

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Abstract

The invention discloses an isolated Methylobacterium sp. DM1 (Methylobacterium sp. DM1), which can decompose N,N-dimethylformamide. Methylobacterium DM1 can use DMF as the only carbon and nitrogen source to grow, completely mineralize it and release ammonia nitrogen. It can tolerate and completely degrade 6800mg / L of DMF in water. Under the conditions of 30°C and pH 7.0, 2000mg / L DMF can be completely degraded within 28 hours, and the degradation rate reaches 71.4mg / L / h, which is one of the fastest reported strains. It is used in environmental sewage treatment and It has a good application prospect in environmental restoration.

Description

Technical field [0001] The present invention relates to the field of environmental biological technology, and more particularly to a N, N-dimethylformamide degradation strain and its application thereof. Background technique [0002] N, N-dimethylformamide (DMF) is a well-known "universal solvent" and is widely used in industrial, agriculture, pharmaceuticals. DMF can be miscible with water and a variety of organic solvents, which have good thermal stability, or a harmful volatile organic compound, which is poorly functioning, and is difficult to be effectively removed from the environment. [0003] Once the DMF enters the environment, it will have a major threat to the health of aquatic organisms and humans. Studies have shown that DMF and its degradation products have significant teratogenic and lethal effects on zebrafish embryos. DMF can enter the human body through the respiratory tract, digestive tract and skin, affecting cell differentiation, causing chromosome distortion,...

Claims

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Application Information

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Patent Type & Authority Patents(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 JIAOTONG UNIV
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