Application of α-ketoglutarate in improving bacterial susceptibility to antibiotics
A technology of ketoglutaric acid and antibiotics is applied in the field of medicine to achieve the effects of overcoming drug resistance, overcoming bacterial drug resistance and improving sensitivity
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Embodiment 1
[0022] Example 1 α-ketoglutarate can increase the sensitivity of bacteria to kanamycin in a concentration-dependent manner
[0023] S1. Preparation of bacterial samples: Pick a single colony of Escherichia coli K12 BW25113 or Edwardsiella tarda EIB202 and culture them in 100 mL of LB or TSB liquid medium at 37°C or 30°C at 200 rpm until saturated. Centrifuge at 8000rpm at room temperature for 2min, collect the bacteria, wash the bacteria three times with sterile saline, and finally adjust the washed bacteria to OD with 1×M9 medium 600 =0.2, draw 5mL of the bacterial solution and distribute it in test tubes for later use.
[0024] S2. Under the premise of 40μg / mL kanamycin, add α-ketoglutaric acid (sterilized by filtration) to make the final concentration of 0, 1.25, 2.5, 5, 10 and 20mM (BW25113) or 0, 0.3, 0.6, 1.2, 2.5 and 5mM (EIB202). Cultivate at 37°C or 30°C, 200rpm for 6 hours, detect the number of viable bacteria with a plate, and calculate the survival rate of differ...
Embodiment 2
[0026] Example 2 α-ketoglutarate can increase the sensitivity of bacteria to various antibiotics
[0027] BW25113 and EIB202 cell samples were prepared according to the method in S1 of Example 1, and different concentrations of different antibiotics were added under the premise of adding α-ketoglutarate at a final concentration of 2.5 mM. For BW25113, add 5 μg / mL gentamicin, 0.5 μg / mL kanamycin, 20 μg / mL streptomycin and 0.5 μg / mL ceftazidime; for EIB202, add 2 μg / mL tetracycline, Chloramphenicol at 200 μg / mL, rifampicin at 12.5 μg / mL, and gentamicin at 16 μg / mL. After incubation at 37°C (BW25113) or 30°C (EIB202) at 200rpm for 6 hours, the number of viable bacteria was detected on a plate, and the survival rate of bacteria with different concentrations of antibiotics was calculated at the concentration of 2.5mM α-ketoglutarate. result( figure 2 ) found that α-ketoglutarate can increase the sensitivity of bacteria to a variety of antibiotics, and the improvement factor is b...
Embodiment 3
[0028] Example 3 α-ketoglutarate can increase the sensitivity of Gram-positive and Gram-negative bacteria to antibiotics
[0029] Select B-hemolytic Streptococcus, Staphylococcus aureus (Gram-positive bacteria) and Pseudomonas aeruginosa (Gram-negative bacteria) to prepare bacterial samples according to the S1 method of Example 1, and add the final concentration to each tube Under the premise of 2.5mM α-ketoglutarate, 8μg / mL gentamicin and 400μg / mL kanamycin (Streptococcus B), and 400μg / mL kanamycin were added and not added, respectively. (Pseudomonas aeruginosa) and 60μg / mL ampicillin (Staphylococcus aureus), after incubation at 37°C and 200rpm for 6h, the number of viable bacteria was detected on the plate, and calculated at 2.5mM α-ketoglutarate , adding different kinds of antibiotics to the survival rate of different kinds of bacteria. by the result ( image 3 ) It can be seen that α-ketoglutaric acid can increase the sensitivity of various bacteria to various antibiotic...
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