Pseudomonas putida WP07, preparation method and purpose
一种恶臭假单胞菌、cgmccno.17760的技术,应用在领域微生物领域,能够解决泔水油成份复杂、工艺过程复杂、成本高等问题,达到实现资源化利用与无害化处理、筛选过程简单、成本低的效果
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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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