Shinella sp. and application thereof to micro-biologically degrading acetaminophen
A technology for acetaminophen and microbial degradation, applied in microorganism-based methods, microorganisms, biological water/sewage treatment, etc., can solve problems such as environmental pollution, major safety risks, DNA strand breaks, etc., and achieve a simple and good preparation process. Application prospect, rapid degradation effect
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Embodiment 1
[0042] Embodiment 1: Screening and identification of strains
[0043] 1) culture medium
[0044] Preparation of inorganic salt medium: NaCl 1g, K 2 HPO 4 1.5g, KH 2 PO 4 0.5g, (NH 4 ) 2 SO 4 1.5g, MgSO 4 0.1g, 1ml trace element solution, made up to 1000ml with deionized water, mixed and stirred evenly, natural pH value, made after high-pressure steam sterilization (121°C, 20min), and each liter of trace element solution contains MnSO 4 ﹒ h 2 O 0.13 g , ZnCl 2 0.23g, CuSO 4 ﹒ h 2 O 0.03g, CoCl 2 ﹒ 6H 2 O 0.42g, Na 2 MoO 4 ﹒ 2H 2 O 0.15g, AlCl 3 ﹒ 6H 2 O 0.05g, make up to 1000ml with deionized water.
[0045] Enriched culture solution: add acetaminophen to the inorganic salt medium so that the final concentration of acetaminophen is 200 mg / L.
[0046] Preparation of LB liquid medium: Yeast powder 10g, peptone 5.0g, sodium chloride 10.0g, deionized water to make up to 1000ml, mixed and stirred evenly, natural pH value, prepared after high-pressure s...
Embodiment 2
[0053] Embodiment 2: the preparation that contains bacterial cell suspension
[0054] (1) Slant culture: Inoculate Shennella HZA2 on the slant medium and culture at 30°C for 3-5 days to obtain the slant; the final concentration of the slant medium is composed of: 10g of yeast powder, 5.0g of peptone, chlorine Sodium chloride 10.0g, agar 20g, deionized water 1000ml;
[0055] (2) Seed culture: pick an inoculation loop from the slant of the bacteria in step (1) and inoculate it into the inorganic salt medium, culture at 30°C for 3-5 days, and obtain the seed liquid; the final concentration of the inorganic salt medium is Composition is with embodiment 1;
[0056] (3) Expansion culture: Inoculate the seed solution obtained in step (2) into LB liquid medium (100 mL) at an inoculum volume concentration of 10-20%, and cultivate it with shaking at 30°C and 150 rpm until the logarithmic growth phase, and obtain Bacterial solution, centrifuge the bacterial solution (6000rpm, 5min), di...
Embodiment 3
[0057] Embodiment 3: Degradation experiment of acetaminophen
[0058] 1) Detection of bacterial concentration and acetaminophen content in inorganic salt medium:
[0059] The growth of bacteria is detected by ultraviolet spectrophotometer, which is expressed by measuring the absorbance value of bacteria in the culture solution at 600nm.
[0060] In this experiment, reversed-phase high-performance liquid chromatography was used to detect the residual amount of acetaminophen in the inorganic salt medium. Reversed-phase high-performance liquid chromatography detection conditions: mobile phase is methanol: H 2 O=15:85 (volume ratio), the analytical column is a Grace Alltima C18 column (4.6×250mm, 5μm), the flow rate is 0.8ml / min, the injection volume is 20μl, and the column temperature is 30°C.
[0061] 2) Paracetamol degradation experiment:
[0062] Take four 250ml Erlenmeyer flasks, add 100ml of inorganic salt culture medium, and add acetaminophen after high-pressure steam st...
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