Sphingomonas paucimobilis strain and application thereof
A technology of less motile sheaths and unicellular bacteria, applied in the field of bioengineering, can solve problems such as non-commercial production, and achieve the effects of convenient operation and simple nutrients
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Embodiment 1
[0031] Example 1 Isolation and screening of Sphingomonas paucimobilis (Sphingomonas paucimobilis) Y5
[0032] Separation: Take the activated sludge sample from the alcohol wastewater plant, make a 10-fold gradient dilution with sterile water, then spread it on the LB agar plate, and culture it at 30°C.
[0033] Primary screening: Pick large, viscous, smooth and moist single colonies for secondary screening.
[0034] Re-screening: The single colonies picked by the primary screening were inoculated in liquid medium (glucose 30g / L, peptone 3g / L, yeast powder 1g / L, K 2 HPO 4 ·7H 2 O2g / L, MgSO 4 ·7H 2 (20.2g / L), 30 DEG C, 200r / min shaker culture, finally screen out the bacterial strain Sphingomonas paucimobilis (Sphingomonas paucimobilis) Y5 that fermented liquid viscosity and crude sugar output are high.
Embodiment 2
[0035] Example 2 Identification of Sphingomonas paucimobilis (Sphingomonas paucimobilis) Y5
[0036] 1) Colony morphology and microscopic examination, the results are as follows figure 2 Shown:
[0037] On the nutrient agar (NA) plate, the colony shape is round, yellow, moist, protruding growth, and the diameter of the colony is 1-5 mm; the microscopic examination shows Gram-negative bacteria.
[0038] 2) Oxidase and contact enzyme test:
[0039] The oxidase test was positive and the contact enzyme test was negative.
[0040] 3) 16srDNA identification:
[0041]Use an inoculation loop to pick 2-5 colonies and transfer them to TE, incubate at 37°C for 1.5h, add NaCl and CTAB solution, mix well, incubate at 65°C for 10min, add phenol-chloroform-isoamyl alcohol solution for extraction, and centrifuge for 10min , transfer the supernatant to a new EP tube, extract twice with chloroform-isoamyl alcohol, transfer to a new EP tube, add isopropanol to precipitate DNA, let stand at ...
Embodiment 3
[0046] Example 3 Detection and analysis of microbial polysaccharides produced by fermentation of Sphingomonas paucimobilis Y5.
[0047] The polysaccharide polymer produced by the fermentation of the strain is soluble in water, but insoluble in organic solvents such as alcohol and ketone.
[0048] The glucuronic acid content measured in the polysaccharide sample by the phenol-sulfuric acid method and the sulfuric acid-carbazole method is 12%; the polysaccharide is composed of D-glucose, D-glucose Aldehydic acid, D-mannose and L-rhamnose composition; Measure mannose in the analysis sample by gas chromatography: glucose: rhamnose=1:3.0:4.5; Determinate glycosidic bond in the polysaccharide by periodic acid oxidation method The types are mainly 1→4 keys and 1→3 keys.
[0049] The above experimental results show that the polysaccharide polymer belongs to welan gum.
[0050] Send the above-mentioned Sphingomonas paucimobilis to the China Type Culture Collection Center for preserva...
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