Acinetobacter lwoffii and application thereof
A kind of technology of Acinetobacter lwedenii and inoculum amount, which is applied in Acinetobacter lwedenii and its application field
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Embodiment 1
[0028] Example 1 Identification of Acinetobacter ruckeri NL1
[0029] (1) Morphological, physiological and biochemical identification of Acinetobacter ruckeri NL1
[0030] Acinetobacter ruckeri NL1 of the present invention is obtained by screening and separating in Yangzhou ancient canal, and its morphological characteristics: in LB solid medium (composition is as follows: yeast extract 5g / L, peptone 10g / L, NaCl 10g / L, pH7 .0-7.2) After 2 days of growth, the colony is milky white, with neat edges, smooth surface, raised, slightly moist, and the diameter of the colony is about 1-1.5 mm, see figure 1 (a). Under the microscope, the bacteria are short rod-shaped, with a length of 2.5-2.9 μm and a width of 1.8-2.1 μm, see figure 1 (b).
[0031] Physiological and biochemical characteristics of Acinetobacter NL1 of the present invention: Gram-negative bacteria; oxidase, catalase negative, can not utilize glucose, lactose, xylose, maltose, sucrose fermentation; escin, citrate, H 2...
Embodiment 2
[0034] Example 2 Tolerance of Acinetobacter lwoffii (Acinetobacter lwoffii) NL1 to phenol
[0035] Inoculate a single colony of Acinetobacter lwoffii (Acinetobacter lwoffii) NL1 stored at 4°C in 20 mL of LB liquid (peptone 10 g, yeast extract 5 g, NaCl 10 g, deionized water to 1 L, adjust pH 7.0) medium Medium, 28°C, 200rpm overnight culture to the logarithmic growth phase, measure the cell concentration, collect the cells, wash twice with PBS buffer and resuspend to OD 600= 1.0. Using this as the initial concentration, carry out 5 times of 10-fold serial dilution, and inoculate 4 μL of the bacterial solution on the solid medium to be tested (solid medium with phenol as the only carbon source), culture at 28°C for 2 days, and observe the bacteria growth and photographed, see image 3 . As the concentration of phenol increased from 0 to 0.5 g / L, the colony density increased, indicating that phenol as a carbon source could support cell growth. With the increase of phenol con...
Embodiment 3
[0037] Embodiment 3,4-aminoantipyridine method measures phenol standard curve
[0038] Accurately weigh 1g of phenol and dissolve it in 1L of deionized water, filter it with a 0.22μm filter membrane to make a 1g / L mother liquor, and then use a pipette to accurately absorb volumes of 0mL, 0.25mL, 0.5mL, 0.75mL, and 1mL , 1.25mL, 1.5mL, 1.75mL, 2mL, 2.25mL, 2.5mL, 2.75mL, and 3mL of mother liquor were distilled to a total volume of sterile deionized water in a 5mL volumetric flask. Take 100 μL of the supernatant in a test tube, add 4.9 mL of distilled water, then add 100 μL of 2mol / L ammonia water, mix well, then add 60 μL of 1% 4-aminoantipyroline (4AT), mix well, and finally add 100 μL of 2% Potassium ferricyanide solution, mix well. After 1 h, at 510 nm, use a cuvette to measure the absorbance of different concentrations of phenol solutions. Phenol standard curve see Figure 4 .
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