Tylosin degrading bacterium as well as screening method and application thereof
A technology of tylosin and screening method, which is applied in the biological field to achieve the effect of efficient degradation
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Embodiment 1
[0021] Example 1. Isolation and screening of tylosin-resistant strains.
[0022] 1) Collect moist soil from the shallow surface (10 cm below the surface) of a free-range chicken farm in Shandong, seal it in a ziplock bag, place it in a low-temperature incubator (the temperature in the box is about 4°C) and bring it back to the laboratory. The chicken farm has been built for about 5 years and uses tylosin as a drug against respiratory infections in poultry. After the obtained soil sample was brought back to the laboratory, 5.0 g was weighed and added to a 500 mL baffle shaker flask containing 100 mL 50 mM sterile phosphate buffer (pH 7.0), and shaken in a constant temperature shaker at 30 ° C for 20 min at a speed of 180 rpm. Take the suspension and dilute it 10 times, 10 times 2 times, 10 3 times, 10 4 Take 100 μL and spread evenly on LB solid medium containing 50mg / L tylosin phosphate (tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L, agar powder 20g / L, 121 ℃ high temperatur...
Embodiment 2
[0032] Example 2. Degradation of Tylosin by P. saccharolyticum TYL-B35.
[0033] The single colonies of P. saccharolyticum TYL-B35 obtained by screening in Example 1 were picked and transferred to 5 mL of LB liquid medium containing 500 mg / L tylosin phosphate, and cultured with shaking at 30° C. and 180 rpm for 18 hours. Pipette 50 μL of bacterial liquid and transfer to 5 mL of LB liquid medium containing 500 mg / L tylosin phosphate. The culture medium was packed in two vials with silica gel stoppers, one with a volume of 25mL (aerobic culture) and the other with a volume of 5mL (facultative anaerobic), which were used to verify aerobic and facultative anaerobic culture respectively. Degradation of tylosin under anaerobic conditions. The inoculated vials were placed in a constant temperature shaker at 30°C and cultured at 180rpm for 5 days. The residual tylosin in the bacterial liquid was determined by HPLC and the degradation rate was calculated. The results showed that the deg...
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