Tetracycline-degrading Bacillus Velez and its magnetically immobilized microbial composite material and its application

A technology for immobilizing microorganisms and Bacillus, applied in the directions of microorganisms, microorganisms, microorganism-based methods, etc., can solve the problems of limited application range, difficult to guarantee the effectiveness of microorganisms, and inconvenient effective recovery of immobilized materials, and achieves the preparation method. Simple, practical application, easy to operate, remove the effect thoroughly

Active Publication Date: 2022-03-25
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technology can not only increase the concentration of microbial cells in a limited area, but also maintain a high activity of microorganisms, but it is difficult to ensure the effectiveness of microorganisms during the immobilization process, and the effective recovery of immobilized materials after use is not convenient, resulting in The scope of application is not wide

Method used

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  • Tetracycline-degrading Bacillus Velez and its magnetically immobilized microbial composite material and its application
  • Tetracycline-degrading Bacillus Velez and its magnetically immobilized microbial composite material and its application
  • Tetracycline-degrading Bacillus Velez and its magnetically immobilized microbial composite material and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Screening, separation, purification and identification of Bacillus velesi that degrades tetracycline

[0052] 1. Screening of Bacillus velesi that degrades tetracycline

[0053] 1. Material preparation

[0054] Soil for strain screening: collected from a grassland in Inner Mongolia Autonomous Region, the soil samples were sealed in ziplock bags, and brought back to the laboratory refrigerator for storage at 4°C for future use.

[0055] LB medium: 10.0 g of peptone, 5.0 g of yeast extract powder, 10.0 g of NaCl, pH 7.0-7.2, dilute to 1 L with distilled water, and sterilize at 121 °C for 20 min.

[0056] LB solid medium: peptone 10.0g, yeast extract powder 5.0g, NaCl 10.0g, agar powder 18.0g, pH 7.0-7.2, dilute to 1L with distilled water, sterilize at 121°C for 20min.

[0057] Mineral salt medium (MSM): 5 mL of phosphate buffered solution (KH 2 PO 4 8.5g / L, K 2 HPO 4 ·H 2 O 21.75g / L, Na 2 HPO 4 ·12H 2 O 33.4g / L, NH 4 Cl 5.0g / L), 3.0mL of MgSO with a ...

Embodiment 2

[0078] Example 2 Degradation of tetracycline by strain N32 in MSM medium containing tetracycline and peptone

[0079] The strain N32 was streaked on an LB plate at 30°C for 24 hours in the dark, and a single colony was picked and inoculated into 20 mL of LB liquid medium, and incubated in a shaker at 30°C and 150 rpm for 18 hours in the dark. The ratio of 2% (v / v) was inoculated into MSM medium containing 10mg / L tetracycline and 10g / L bacteriological peptone for dark culture, and samples were taken after 3 and 6 days; MSM medium without strain N32 was inoculated. as the control (CK). After the bacterial liquid was filtered, the residual amount of tetracycline TC was measured by HPLC (the tetracycline determination method was the same as that in Example 1).

[0080] The results showed that the control tetracycline that was not inoculated with strain N32 was hydrolyzed after 3 days, and the degradation rate of tetracycline increased after inoculation with strain N32; 6 days lat...

Embodiment 3

[0081] Example 3 Preparation of magnetically immobilized microbial composite material based on strain N32 and verification of its ability to adsorb and degrade tetracycline

[0082] 1. Preparation of Magnetic Immobilized Microbial Composites

[0083] (1) Mix the sodium alginate and ultrapure water, stir magnetically until the dissolution is complete, and obtain a sodium alginate solution with a concentration of 10% (g:mL). CaCl 2 Mix with ultrapure water, stir magnetically until dissolved completely, get CaCl 2 solution, the concentration is 8%.

[0084] (2) sodium alginate solution, CaCl obtained to step (1) 2 Add hydrochloric acid (concentration: 6mol / L) to the solution respectively, adjust the pH to 7, put it in an autoclave, sterilize at 121°C for 20min, take out and cool to room temperature.

[0085] (3) After streaking the strain N32 on an LB plate at 30°C for 24 hours in the dark, pick a single colony and inoculate it in 20 mL of LB liquid medium, and inoculate it i...

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Abstract

The invention discloses a tetracycline-degrading Bacillus Velez and a magnetically immobilized microbial composite material and its application. The name of the bacterium is Bacillus velezensis N32, and it was preserved on November 6, 2019 at the Guangdong Provincial Microbiological Culture Collection Center located on the 5th Floor, Building 59, Compound No. 100, Xianlie Middle Road, Guangzhou, with the preservation number For GDMCC NO:60887. The bacterium has the ability to degrade tetracycline, and can be applied to the degradation of tetracycline in soil and water, so as to remove or reduce the residue of tetracycline in the environment and achieve the effect of controlling environmental pollution. The magnetically immobilized microbial composite material prepared by the bacterium can improve the tetracycline degradation effect of Bacillus Velez N32, the tetracycline removal effect is more thorough, and it is easy to recycle, and the actual use is more convenient and environmentally friendly.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, organic pollution microbial restoration and removal of environmental organic pollutants such as water bodies and soils, and particularly relates to a tetracycline-degrading Bacillus velesi and magnetic immobilized microbial composite materials and applications. Background technique [0002] Antibiotics are essential drugs in agricultural production and aquaculture, and are also one of the most widely used human medical drugs in the world. However, long-term use will inevitably lead to drug residues and cause serious environmental pollution problems. China is the world's largest producer and user of antibiotics. It is estimated that in 2013, the total use of 36 antibiotics in my country reached 92,700 tons, and about 53,800 tons of antibiotics entered the environment, causing serious environmental pollution. Among them, tetracycline antibiotics Pollution is the worst. Due to the high solub...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N11/10C12N11/14C02F3/34B09C1/10C02F101/34C02F101/38C12R1/07
CPCC12N11/10C12N11/14C02F3/348B09C1/10C02F2101/34C02F2101/345C02F2101/38C12R2001/07C12N1/205
Inventor 徐会娟李永涛郭桂芳吴祥瑶张玉龙龚贝妮谭泽文刘永林
Owner SOUTH CHINA AGRI UNIV
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