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Screening and identification and application of bacillus for efficiently degrading phthalate

A technology for Bacillus subtilis and strains, which is applied in the field of screening and identification of efficiently degrading Bacillus phthalates, and can solve the problems of refractory degradation, slow rates of hydrolysis and photolysis, and long half-life of phthalates.

Active Publication Date: 2014-02-12
SICHUAN AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Phthalates have a long half-life in the environment, and their hydrolysis and photolysis rates are very slow, so they are refractory substances. Microbial degradation of PAEs is a natural environment The main way is currently only the patent of Gordonia sp.QH-9 degradable phthalates reported by Yu Zhisheng et al.

Method used

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  • Screening and identification and application of bacillus for efficiently degrading phthalate
  • Screening and identification and application of bacillus for efficiently degrading phthalate
  • Screening and identification and application of bacillus for efficiently degrading phthalate

Examples

Experimental program
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Effect test

example 1

[0027] Example 1: Determine the relationship between the growth of phthalate-degrading bacterial strain JF and the degradation of dibutyl phthalate

[0028] Activation of strains: pick the strain JF that degrades phthalates into LB liquid medium, 35°C, 180r / min shaking culture for 24h; then continuously streak, pick a single colony and culture twice (35°C), Pick a single colony and inoculate it on an LB slope at 35°C for 2 days for later use.

[0029] Wash the bacterial cells on the lower slant with sterile saline and adjust the cell concentration to 10 8 cfu / mL, made into seed solution.

[0030] Determination of the growth curve: Inoculate the seed liquid of the degrading strain JF into LB liquid medium containing 200 mg / L dibutyl phthalate (DBP) according to the inoculum amount of 5%, and divide into 30 mL / 250 mL Erlenmeyer flasks, Cultivate at 35°C and 180r / min for 48h. The sampling time is 1 bottle every 2h during 0h-36h, and 1 bottle every 4h during 36h-48h. Store the s...

example 2

[0033] Example 2: Degradation characteristics of strain JF degrading dibutyl phthalate

[0034] Effect of substrate concentration on the degradation rate of dibutyl phthalate by strain JF: According to the inoculum amount of 5.0% (v / v), the seed solution (1.0×10 8 individual / mL) inoculated in LB liquid medium (pH7.0, 100 mg / L, 200 mg / L, 300 mg / L, 400 mg / L) with different concentrations of dibutyl phthalate, and aliquoted 30 mL / 250 mL Erlenmeyer flask, 35°C, 180 r / min shaking culture, sampling interval 4h. Use [0024] described method to measure dibutyl phthalate degradation rate.

[0035] Effect of culture temperature on the degradation rate of dibutyl phthalate by strain JF: According to the inoculation amount of 5.0% (v / v), the seed solution (1.0×10 8 / mL) inoculated in LB liquid medium (pH7.0) with a dibutyl phthalate mass concentration of 200 mg / L, divided into 30 mL / 250 mL Erlenmeyer flasks, at different temperatures (30°C, 35°C, 40°C, 45°C) 180 r / min shaking culture,...

example 3

[0038] Example 3: Degradation effect of phthalate-degrading strain JF on dibutyl phthalate in MM medium

[0039] Inoculate the seed solution of the degraded strain into 200 mg / L dibutyl phthalate (DBP) MM liquid medium according to the inoculum amount of 5%, and divide into 30 mL / 250 mL Erlenmeyer flasks, and inoculate 5% sterile water As a blank control, 35 ℃, 180r / min shaking culture.

[0040] Sampling after cultivating 60h, measure the residual amount of dibutyl phthalate (DBP) in the nutrient solution with the method described in [0024], control group dibutyl phthalate (DBP) hardly degrades, in the experimental group about 92.66% of dibutyl phthalate (DBP) was degraded by strain JF.

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Abstract

The invention provides a bacterial strain JF which is separated out from a large number of soils with mulching films used and can efficiently degrade phthalate, and belongs to the technical field of biology. The bacterial strain grows fast, can produce spore, is high in resistance to external environment, and reaches more than 90% of degradation rate regarding to phthalate within a broad range that the temperature is 30 to 45 DEG C and the pH (Potential of Hydrogen) is 5.0 to 8.0. The identification result shows that the bacterial strain is bacillus subtilis (Bacillus Subtilis), and the accession number is KF364634; and the bacterial strain is stored in China General Microbiological Culture Collection Center (CGMCC) located at No.3, Courtyard 1, West Beichen Road, Chaoyang District, Beijing, on July 22, 2013, with the storage number of CGMCC No.7950. The bacterial strain can be applied to bioremediation or biological purification of soil and sewage polluted by phthalate, and protects the ecological environment, and thus the safety of agricultural products is ensured.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the screening and identification of a highly efficient phthalate-degrading bacillus and its application. Background technique [0002] Phthalates (Phthalates, PAEs, phenolphthalein esters) are one of the most widely produced and widely used synthetic organic compounds in the world. They are aromatic dicarboxylic acid esters, mostly with high boiling point, low vapor pressure, They are widely used as plasticizers and softeners in plastic additives, paint solvents, synthetic rubber and coatings to increase the plasticity and toughness of plastics and improve the strength of plastics. PAEs compounds are a general term for about 30 compounds, which are composed of phthalic acid and corresponding alcohols. The commonly used varieties in my country are: dibutyl phthalate (DBP), bis(2-phthalate) Ethylhexyl (DEHP), dioctyl phthalate (DOP), butyl benzyl phthalate (BBP), dinonyl phthalate (DNP...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34B09C1/10C12R1/125C02F101/38
Inventor 不公告发明人
Owner SICHUAN AGRI UNIV
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