Pseudomonas putida WP07, preparation method and purpose
A technology of Pseudomonas putida, cgmccno.17760, applied in the field of microorganisms, can solve the problems of complex swill oil composition, complex process and high cost, achieve resource utilization and harmless treatment, simple screening process, low cost effect
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Embodiment 1
[0036] Embodiment 1: Domestication and screening of Pseudomonas putida WP07 bacterial strain
[0037] The soil from the smoke exhaust outlet of the canteen was selected, and the Pseudomonas strains that transformed and synthesized PHA using swill oil as a single carbon source were obtained through domestication with high concentration of swill oil and selective selection with Nile blue. Strain acclimatization and screening steps include:
[0038] (1) Acclimatization of strains: first, add swill oil to the inorganic salt medium at a ratio of 20g / L, then obtain a soil sample from the soil at the smoke exhaust outlet of the canteen and add it to the above-mentioned inorganic medium at a ratio of 5g / L. Cultivation and acclimatization; the acclimatization conditions of the strain are temperature 20°C-30°C, rotation speed 200-300rpm; every 5 days, continue to add swill oil to the inorganic salt medium at a ratio of 5g / L until the swill oil in the inorganic salt medium is added Afte...
Embodiment 2
[0044] Example 2 Effect of Fermentation Temperature on Pseudomonas putida WP07 Transformation and Synthesis of PHA Efficiency
[0045] Insert the Pseudomonas putida WP07 obtained in Example 1 into the seed culture medium, the inoculum size is 1% (V / V), at 30°C, 200rpm number of revolutions conditions are cultivated for 24h activation to obtain the activated Pseudomonas seed culture.
[0046] Add swill oil as a single carbon source to the inorganic salt medium at a ratio of 20g / L, and add 1:10 (w / w) tween 80 as an emulsifier at the same time; then the activated Pseudomonas seed culture solution Insert the inoculation amount of 3% (v / v) into the inorganic salt medium added with swill oil, control the culture temperature at 20°C, 25°C, 30°C, 35°C and 40°C, and cultivate under the condition of 300rpm 72h (result comparison see figure 1 ). Among them, the seed medium preparation method is to add 10g / L tryptone, 5g / L yeast extract and 10g / L sodium chloride to deionized water, adj...
Embodiment 3
[0051] Example 3 Influence of the pH value of the inorganic salt medium on the conversion and synthesis of PHA by Pseudomonas putida WP07
[0052] The Pseudomonas putida WP07 obtained in Example 1 was inoculated into the seed culture medium, and cultured for 24 hours at 30° C. and 200 rpm for activation to obtain an activated Pseudomonas seed culture solution.
[0053] Add swill oil as a single carbon source to the inorganic salt medium at a ratio of 20g / L, and add 1:10 (w / w) tween 80 as an emulsifier at the same time; then add the seed culture solution to 3% (v / v ) was inserted into the inorganic salt medium added with swill oil, and the culture temperature was controlled to be 30° C. and the rotation speed was 300 rpm to cultivate for 72 hours. Among them, the seed medium preparation method is to add 10g / L tryptone, 5g / L yeast extract and 10g / L sodium chloride to deionized water, adjust the pH value with 0.1N HCl and 0.1N NaOH solution after mixing is 7.0.
[0054] Inorgan...
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