A β-lactam antibiotic degrading strain pgb1 and its application
A technology of lactams and antibiotics, applied in the field of microorganisms
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Embodiment 1
[0024] 1. Screening of β-lactam antibiotics degrading strain PGB1
[0025] 1. Material preparation
[0026] The source of bacteria samples: fresh bacteria residues from β-lactam antibiotics manufacturers.
[0027] Beef extract-peptone medium: 5.0g beef extract, 10.0g peptone, 5.0g NaCl, distilled water, pH 7.4-7.6, sterilized at 21°C for 15min. Add 10g-15g of agar to the solid medium.
[0028] Martin's Medium: KH 2 PO 4 1.0g, MgSO 4 7H 2 O 0.5g, glucose 10.0g, peptone 5.0g, distilled water 1L, pH natural, sterilized at 115°C for 15min. Add 10g-15g of agar to the solid medium.
[0029] Inorganic salt medium: (NH 4 ) 2 NO 4 1.0g, K 2 HPO 4 0.5g, KH 2 PO 4 0.5g, MgSO 4 ·7H 2 O0.2g, NaCl0.2g, CaCl 2 0.1g, MnSO 4 h 2 O 0.01g, FeSO 4 ·7H 2 O 0.01g, distilled water 1L, pH7.0, sterilize at 121℃ for 15-20min. Add 10g-15g of agar to the solid medium.
[0030] 2. Strain screening
[0031] (1) Strain isolation and purification
[0032] Separation and purificati...
Embodiment 2
[0088] Growth and degradation characteristics of strain PGB1
[0089] 1. Strain PGB1 grows in penicillin potassium medium
[0090]The bacterial strain PGB1 was inoculated in beef extract peptone liquid medium for activation culture. According to the inoculation amount of 3-6%, the activated bacterial liquid was taken, centrifuged at 8000r / min for 5min to obtain bacterial cells, and then inoculated to 50 -1000mg / L beef extract peptone solid and liquid medium with different concentrations of penicillin potassium, culture the solid plate at 28--37°C, observe the colony size regularly; place the liquid medium at 28--37°C, Shake culture at 150-180r / min, take samples regularly, measure the OD value at 600nm, see the results image 3 .
[0091] The results showed that strain PGB1 could grow well in beef extract peptone liquid and solid medium containing 50-1000 mg / L penicillin potassium. 8-10h enters the logarithmic growth phase, and the biomass reaches the maximum at 48h.
[009...
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