Phosphorus-dissolving pseudomonas putida L13 and fermentation process thereof
A technology of Pseudomonas putida and phosphorus solubilization, applied in the field of microorganisms, can solve the problems of complex fermentation production process and narrow practical application range, and achieve strong application prospects, strong environmental adaptability, and growth-promoting effects
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Embodiment 1
[0058] Embodiment 1 Shake flask fermentation culture
[0059] A. First-class seed preparation: take the slanted seeds of Pseudomonas phospholytica putida L13 (preservation number is CCTCC NO: 2010342) and draw a line on the LB medium plate, put it into a 28° C. incubator for 14 hours, and free single colonies appear. This is the primary seed;
[0060] B. Preparation of secondary seeds: use an inoculation loop to take a single colony of the primary seed of step A, and insert it into a 250mL shake flask with 50mL of LB liquid medium, control the temperature at 28°C, and the shaker speed at 180rpm, and ferment and cultivate 12h, OD 600 Reaching 2.0, this is the second-level seed;
[0061] C, fermentation culture: get the secondary seed liquid of step B to insert in the 250mL shaking flask that 30mL fermentation medium is housed with the inoculum size of 2% (V / V) and carry out fermentation culture, control shaker temperature to be 28 ℃, rotating speed 180rpm, fermentation cultu...
Embodiment 2
[0066] Embodiment 2 Shake flask fermentation culture
[0067]A. First-class seed preparation: take the slanted seeds of Pseudomonas phospholytica putida L13 (preservation number is CCTCC NO: 2010342) and draw a line on the LB medium plate, put it into a 28° C. incubator for 14 hours, and free single colonies appear. This is the primary seed;
[0068] B. Preparation of secondary seeds: use an inoculation loop to take a single colony of the primary seed of step A, and insert it into a 250mL shake flask with 50mL LB liquid medium, control the temperature at 30°C, and shake the table at a speed of 200rpm, and ferment and cultivate 14h, OD 600 Reaching 2.0, this is the second-level seed;
[0069] C, fermentation culture: get the secondary seed liquid of step B to insert in the 250mL shaking flask that 40mL fermentation medium is housed with the inoculum size of 3% (V / V) and carry out fermentation culture, control shaker temperature to be 30 ℃, rotating speed 200rpm, fermentation...
Embodiment 3
[0074] Embodiment 3 Shake flask fermentation culture
[0075] A, primary seed preparation: take the slant seeds Pseudomonas putida L13 (preservation number is CCTCC NO: 2010342) and draw a line on the LB medium plate, put it into a 28°C incubator and cultivate it for 10h, free single colonies appear, This is the primary seed;
[0076] B. Preparation of secondary seeds: use an inoculation loop to take a single colony of the primary seed of step A, and insert it into a 250mL shake flask with 30mL LB liquid medium, control the temperature at 26°C, and shake the table at a speed of 160rpm. Fermentation 10h, OD 600 Reaching 2.0, this is the second-level seed;
[0077] C, get the secondary seed liquid of step B to insert in the 250mL shaking flask that 20mL fermentation medium is housed with the inoculum size of 0.5% (V / V) and carry out fermentation culture, control shaker temperature is 26 ℃, and rotating speed is 160rpm, Fermentation culture to 36h, OD 600 reached 15.0;
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