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Bacillus subtilis L1, and application thereof in degradation of lincomycin residual in lincomycin residues

A technology of Bacillus subtilis and lincomycin, which is applied to bacteria, microorganism-based methods, microorganisms, etc., can solve the problems of increasing difficulty in the composition of antibiotic bacteria residues, secondary pollution, and rapid decay of bacteria residues.

Active Publication Date: 2018-12-18
龙海市农丰源肥料有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The complexity of the composition of antibiotic residues increases the difficulty of solid fermentation; the residual antibiotics in the residues inhibit the growth of fermentation microorganisms; the residues decompose quickly, the solid fermentation time is long, and the odor is serious during the fermentation process, resulting in secondary pollution

Method used

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  • Bacillus subtilis L1, and application thereof in degradation of lincomycin residual in lincomycin residues
  • Bacillus subtilis L1, and application thereof in degradation of lincomycin residual in lincomycin residues
  • Bacillus subtilis L1, and application thereof in degradation of lincomycin residual in lincomycin residues

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Screening and identification of embodiment 1 bacterial classification

[0027] 1. Strain screening

[0028] Broth agar medium mass final concentration composition: peptone 1%, beef extract 0.3%, sodium chloride 0.5%, agar 2%, solvent is water, pH value 7.4.

[0029] Dilute the sewage obtained from the aerobic tank of Henan Tianfang Pharmaceutical Wastewater Treatment Plant to different times (2.5 times, 25 times, 250 times, 2500 times and 25000 times respectively), and add lincomycin to make each dilution The final concentrations of lincomycin in high-degree sewage were 0.6 g / L and 6 g / L, respectively, and a series of sewage lincomycin mixtures were prepared.

[0030] Pour 20 mL broth agar medium into each Petri dish while hot to make a plate. The above sewage lincomycin mixture was then spread onto the plates. After culturing at 37°C for 12 h, four strains were screened out according to the colony morphology, which were designated as strain L1, strain L2, strain L3,...

Embodiment 2

[0063] Embodiment 2 strain culture

[0064] The bacterial strain L1-strain L4 screened out in Example 1 were inserted into common broth agar medium respectively, and cultured at 37° C. for 24 hours to obtain slant culture for later use; the slant seeds were inoculated into common broth culture, and the Cultivate at 37°C and 200 r / min for 12 hours to obtain seeds. Seeds were inoculated at 10% (v / v) in common broth medium, cultured at 37°C and 200 r / min, and samples were taken regularly. The proliferation of the strain was observed under a microscope and the absorbance at 600 nm was measured. The growth curve is shown in figure 2 . It can be seen from the figure that the logarithmic phase of these strains appeared at different times, L1 was 4-12 h, L2 was 4-10 h, L3 was 6-12 h, and L4 was 20-28 h.

Embodiment 3

[0065] The establishment of embodiment 3 lincomycin standard curve

[0066] Stationary phase: octadecylsilane bonded silica gel (5 µm) column for liquid chromatography, 4.6 mm×200 mm, mobile phase: 0.05M borax solution (adjust pH to 6.1 with 85% phosphoric acid solution) and methanol Mix (volume ratio 42:58), flow rate: 1 mL / min. Column temperature: 30°C, detector: ultraviolet detector, detection wavelength: 214 nm.

[0067] Under the above conditions, within the range of 0-8.0 mg / mL, the concentration (x) of lincomycin standard aqueous solution has a linear relationship with the ultraviolet absorption (peak area, y), and the linear regression equation is y = 1.1013x –0.0203, R 2 = 0.9997.

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Abstract

The invention discloses a Bacillus subtilis L1, and an application thereof in the degradation of lincomycin residual in lincomycin residues. The application is characterized in that the lincomycin residues are inoculated with a fermentation solution obtained by fermenting and culturing the Bacillus subtilis L1, and are cultured at 25-37 DEG C under 80-220 rpm to realize the degradation of lincomycin in the residues. The Bacillus subtilis L1 can degrade the lincomycin residual in lincomycin residues, and the lincomycin degradation rate within 6 days reaches 95.39%; and microorganisms for degrading antibiotics also can be used for degrading antibiotics in tap water or soil.

Description

technical field [0001] The invention relates to a strain and a method for harmlessly treating lincomycin in lincomycin fermentation residue, belonging to the field of environmental protection. Background technique [0002] my country is a major producer of antibiotics, with an annual output of more than 140,000 tons of major antibiotics, and an annual output of more than 1.4 million tons of solid waste (mycelium and protein), namely antibiotic residues, produced during antibiotic production. [0003] Antibiotic residues are solid fermentation wastes produced in the process of fermenting and producing antibiotics. A small amount of antibiotics is a special kind of hazardous waste. If it is not disposed of properly, it will cause potential harm to the ecological environment and human health. The harm has the characteristics of concealment, lag, accumulation, synergy and jointness. [0004] There are a small amount of residual antibiotics and their degradation products in anti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A62D3/02C12R1/125A62D101/28
CPCA62D3/02A62D2101/28C12N1/205C12R2001/125
Inventor 李永红付路平韩正李晓静刘颖颖许福兰
Owner 龙海市农丰源肥料有限公司
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