A kind of sphingomonas paucimobilis and its application
A technology of oligokinesis and monocysts, applied in the field of degrading bacteria, to achieve the effect of strong environmental adaptability and simple nutritional requirements
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Embodiment 1
[0026] Embodiment 1: Screening of bacterial strain YZ-4
[0027] 1. Culture medium:
[0028] Enrichment medium composition: peptone 10g / L, yeast powder 5g / L, glucose 20g / L, edible blend oil 5g / L, 6g / L Tween 80, solvent is deionized water, pH is natural. Sterilize at 115°C for 30 minutes.
[0029] The composition of acclimation medium: dipotassium hydrogen phosphate 1g / L, potassium dihydrogen phosphate 1g / L, disodium hydrogen phosphate 1g / L, sodium chloride 0.5g / L, magnesium sulfate heptahydrate 0.5g / L, solvent is deionized Water, pH natural. Sterilize at 121°C for 20 minutes.
[0030] Oil screening medium composition: ammonium sulfate 2g / L, dipotassium hydrogen phosphate 1g / L, potassium chloride 0.5g / L, magnesium sulfate heptahydrate 0.5g / L, ferrous sulfate 0.01g / L, agar 20g / L, Edible blended oil emulsion 12mL / L, solvent is deionized water, pH is natural. Sterilize at 121°C for 20 minutes. The edible blended oil emulsion is obtained by emulsifying the edible blended oil ...
Embodiment 2
[0040] Embodiment 2: Identification of bacterial strain YZ-4
[0041] 1. Colony morphology observation:
[0042] Use an inoculation loop to pick a small amount of preserved strains from the slant of Example 1, streak on the YPD plate, and culture at 37°C for 1-2 days. The colony morphology is shown in figure 1 . The colony is yellow, protruding, with a smooth and round surface.
[0043] 2. SEM scanning electron microscope observation:
[0044] Cells are rod-shaped, without capsule, flagella, spores ( figure 2 ).
[0045] 3. Molecular identification:
[0046] Template preparation: pick a single colony in a medium containing 20 μL sterile ddH 2 In the EP tube of O, cook in boiling water for 10 minutes, centrifuge at 12,000 rpm for 1 minute, and use the supernatant as a DNA template. 16S rDNA sequence was amplified by PCR with 27F (5'-AGAGTTTGATCCTG GCTCA-3') and 1492R (5'-AAGGAGGTGATCCAGCCGCA-3') as forward and reverse primers.
[0047] PCR amplification system (50 μL):...
Embodiment 3
[0062] Example 3: Application of Sphingomonas paucimobilis YZ-4 in degrading edible blended oil
[0063] 1. Culture medium:
[0064] With embodiment 1.
[0065] 2. Experimental method:
[0066] Pick a ring of the strain YZ-4 preserved on the slant and place it in a test tube containing 5mL LB liquid medium, culture at 37°C and 200rpm for 12-14h to obtain seed liquid; transfer 1mL of seed liquid to 100mL LB liquid culture medium In a 500mL Erlenmeyer shake flask based on the base, culture at 37°C and 150rpm for 12-14h to obtain a fermentation broth. The fermentation broth was centrifuged at 4°C and 8000 rpm for 10 min, and the wet cells were collected for later use.
[0067] pH optimization: adjust the pH (4, 5, 6, 7, 8, 9) of the inorganic salt medium with HCl and NaOH respectively, and inoculate the wet bacteria into the inorganic salt containing 40g / L edible blending oil according to the inoculation amount of 10g / L In the culture medium, react for 7 days at 37°C and 150r...
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