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Constitutive 1,4-dioxane-degrading bacterium

A dioxane and compositional technology, applied in bacteria, biological water/sewage treatment, organic chemistry, etc., can solve the problem of low specific degradation rate and achieve low-cost effects

Inactive Publication Date: 2018-03-27
TAISEI CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] However, as described in Non-Patent Document 3, there is a problem that the maximum specific degradation rate of 1,4-dioxane of constitutive 1,4-dioxane-degrading bacteria is lower than that of inducible 1,4-dioxane-degrading bacteria
In addition, it is unknown whether the 1,4-dioxane-degrading bacteria disclosed in Patent Document 1 and Non-Patent Documents 3 and 4 can degrade other cyclic ethers in the presence of 1,4-dioxane

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] "Determination of the 16S rDNA partial base sequence of the N23 strain"

[0062] The N23 strain was cultured for 7 days (28° C., 120 rpm) in a CGY liquid medium (5 g / L of casetone (Casitone), 5 g / L of glycerol, and 1 g / L of yeast extract). The culture solution was centrifuged at 10,000×g at 4° C. for 3 minutes to collect bacteria, and washed twice with 0.9% physiological saline. From the cleaned cells obtained, according to "Co-translated by Kaoru Saigo and Yumiko Sano: Molecular Biology Experiment Operation Procedure I Adjustment of Bacterial Genomic DNA Basic Operation Procedure Small Adjustment of Bacterial Genomic DNA (miniprep), pp. 36-37, Maruzen Co., Ltd., 1997. "The DNA was extracted, and the PCR amplification of 16S rDNA was performed. As primers, 8F (5'-AGAGTTTGATCCTGGCTCAG-3') and U1492R (5'-GGTTACCTTGTTACGACTT-3') were used. PCR amplification was performed at 94°C for 10 minutes, followed by 35 cycles of denaturation (94°C, 1 minute), annealing (58°C, 1 mi...

Embodiment 2

[0066] "Investigation on the degradation characteristics of 1,4-dioxane of N23 strain"

[0067] Add 100 mL of CGY medium (5 g / L casein, 5 g / L glycerol, 1 g / L yeast extract) to a 300 mL capacity Erlenmeyer shaker flask with baffles, and sterilize in an autoclave (121°C ,15 minutes). After that, 1,4-dioxane was added to make it 500 mg / L, and the N23 strain was inoculated with bacteria in the amount of one inoculated loop, and subjected to rotary shaking culture (28°C, 120rmp) for 7 days (pre-pre-culture ).

[0068] After the culture, subculture was carried out in a CGY medium containing 500 mg / L of 1,4-dioxane, and culture was carried out under the same conditions (pre-culture).

[0069] The culture solution obtained in the pre-cultivation was centrifuged to collect and recover bacteria, and an inorganic salt medium (composition: 1g / L K 2 HPO 4 , 1g / L (NH 4 ) 2 SO 4 , 50mg / L NaCl, 200mg / L MgSO 4 ·7H 2 O, 10mg / L FeCl 3 , 50mg / LCaCl 2 , pH: 7.3) to clean the bacteria. ...

Embodiment 3

[0074] "Confirmation of proliferation of N23 strain by 1,4-dioxane"

[0075] After adding 1 mL of the N23 strain bacterial inoculum prepared in Example 2 to 19 mL of inorganic salt medium, 1,4-dioxane was added to make it a predetermined concentration, and rotation shaking culture was carried out at 28° C. and 120 rpm (n = 3). The culture was carried out for 6 hours, and the 1,4-dioxane concentration before and after the culture was measured by headspace GC / MS, and the protein concentration was measured by the method shown in Example 2. The ratio of the increased protein amount to the amount of 1,4-dioxane degradation was calculated as the cell yield. Figure 5 Indicates the relationship between initial 1,4-dioxane concentration and cell yield.

[0076] An increase in the amount of protein was confirmed in all the experimental systems, and it was confirmed that the N23 strain propagated using 1,4-dioxane as a carbon source. In particular, in the experimental system with an ...

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Abstract

The present invention addresses the problem of providing a constitutive 1,4-dioxane-degrading bacterium having an excellent maximum specific 1,4-dioxane degradation rate. As a solution, a constitutive1,4-dioxane-degrading bacterium is provided, which is a bacterium of strain N23 that has been deposited under the deposition number NITE BP-02032.

Description

technical field [0001] The invention relates to a constitutive 1,4-dioxane degrading bacterium. Background technique [0002] 1,4-Dioxane is a cyclic ether represented by the following formula (1). 1,4-Dioxane has excellent compatibility with water and organic solvents, and is mainly used as a reaction solvent for organic synthesis. [0003] [0004] In 2010, the production and import volume of 1,4-dioxane in Japan was about 4500t / year, and it is estimated that about 300t / year was discharged into the environment. Since 1,4-dioxane is water soluble, it will spread widely if discharged into the aquatic environment. In addition, it is difficult to remove from water because of low volatility, solid adsorption, photodegradability, hydrolyzability, and biodegradability. Since 1,4-dioxane has been pointed out to be carcinogenic in addition to its acute toxicity and chronic toxicity, there is concern that pollution of the water environment by 1,4-dioxane will have adverse effe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C02F3/00C12N1/00
CPCB09C1/10C02F3/00C12N1/20A62D3/02A62D2101/28C07D319/10C12R2001/01C12N1/205C02F3/34C07C43/13
Inventor 山本哲史斋藤祐二泷宽则
Owner TAISEI CORP
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