A kind of separation method and application of aminoglycoside antibiotic degrading bacteria fn-a2

An aminoglycoside, FN-A2 technology, applied in microorganism-based methods, biochemical equipment and methods, separation of microorganisms, etc., can solve problems such as difficult degradation and harm, and achieve the effects of low cost and simple operation.

Active Publication Date: 2019-06-14
FUJIAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, aminoglycoside antibiotics have a stable chemical structure, heat resistance, acid and alkali resistance, and are difficult to degrade. Their toxicity also has a concentration accumulation effect, which is seriously harmful.

Method used

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  • A kind of separation method and application of aminoglycoside antibiotic degrading bacteria fn-a2
  • A kind of separation method and application of aminoglycoside antibiotic degrading bacteria fn-a2
  • A kind of separation method and application of aminoglycoside antibiotic degrading bacteria fn-a2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1. Screening and enrichment of aminoglycoside antibiotic-degrading strains with increasing concentration gradient

[0029] Collect 50 grams of soil samples, dilute with 100ml sterile water, mix well, take the suspension and inoculate it into the LB culture medium with 300mg / L aminoglycoside antibiotics after standing still, and culture at 37°C and 180r / min constant temperature shaking for 3 After the culture became turbid, the culture was transferred step by step to the LB culture solution with a concentration of 600 mg / L, 900 mg / L and 1200 mg / L aminoglycoside antibiotics every 3 days, and the concentration gradient was gradually screened and enriched. , resulting in a turbid high-concentration target culture.

[0030] 2. Isolation and purification of aminoglycoside antibiotic degrading strains

[0031] Put the high-concentration target culture obtained in step 1 (1) into 1 / 10 concentration LB culture solution containing 1000 mg / L aminoglycoside antibiotics for culture...

Embodiment 2

[0039] The concentration gradient screening of aminoglycoside antibiotic-degrading strains, re-screening and purification of aminoglycoside antibiotic-degrading strains, and the preparation process of the degrading bacteria seed solution are the same as in Example 1.

[0040] 2 Dingsuliang 5383 Take 1000g of contaminated soil containing 800 mg / L of aminoglycoside antibiotic streptomycin, insert 100mL of the seed solution of degrading bacteria FN-A2, mix thoroughly, and replenish water until the total water content is controlled at about 60%. After 90 hours of fermentation, the bioactivity analysis method detected that the residual antibiotics in the treated soil had been basically degraded, and the degradation rate reached more than 98%.

Embodiment 3

[0042] The concentration gradient screening of aminoglycoside antibiotic-degrading strains, re-screening and purification of aminoglycoside antibiotic-degrading strains, and the preparation process of the degrading bacteria FN-A2 seed solution are the same as in Example 1.

[0043] 2 Dingsuliang 5383 Connect 10m of the seed solution of the above-mentioned degrading bacteria FN-A2 to 100 mL of sewage containing 1000 mg / L aminoglycoside antibiotic gentamycin, culture at a constant temperature of 37°C and 200r / min, and analyze the biological activity According to the detection method, the antibiotics in the water body decreased to 20% in 15 hours, 84.40% in the water body in 28 hours, and 96.7% in the water body in 40 hours.

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Abstract

The invention relates to a separating method of aminoglycoside antibiotics degrading bacteria FN-A2 and application of the degrading bacteria.According to the technical scheme, a soil sample is collected to prepare turbid liquid, and bacteria are inoculated into an LB culture solution containing aminoglycoside antibiotics in a mode that the concentration is gradually increased in gradient to be subjected to enrichment culture and separated into strains of different morphologies; the obtained strains of different colonial morphologies are independently screened with a sterile aqueous solution only containing aminoglycoside antibiotics with concentration of 1000 mg / L to obtain aminoglycoside antibiotics degrading bacteria FN-A2.The degrading bacteria FN-A2 are subjected to molecular identification and named as Paracoccus kondratievae FN-A2.The strains can rapidly degrade aminoglycoside antibiotics such as kanamycin, streptomycin, gentamicin and tobramycin in sewage, soil or bacterium dregs or can be used for biological repair of a water body and soil polluted by aminoglycoside antibiotics.

Description

technical field [0001] The invention relates to the fields of antibiotic fermentation and environmental protection, in particular to a method for separating aminoglycoside antibiotic-degrading bacteria FN-A2 and its application. Background technique [0002] Aminoglycoside antibiotics are a class of antibiotics linked by amino sugars and aminocycloalcohols through oxygen bridges, mainly produced by actinomycetes. Its mechanism of action is to inhibit the synthesis of cellular proteins, and its target is the A site of the 16S rRNA decoding region of the 30S small subunit of the ribosome, hindering the synthesis of the initial complex, inducing cells to synthesize wrong proteins and inhibiting the release of synthesized proteins, resulting in Bacteria die. It is mainly used clinically for systemic infections caused by sensitive Gram-negative aerobic bacilli, such as meningitis, respiratory tract, urinary tract, gastrointestinal tract, skin and soft tissue, burns, trauma, and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C02F3/34B09C1/10C12R1/01C02F101/38
CPCB09C1/10C02F3/34C02F2101/38C12N1/02C12N1/20
Inventor 王明兹陈立文方森海吕月霞张会吴钦缘陈必链
Owner FUJIAN NORMAL UNIV
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