Pseudomonas aeruginosa and application thereof
A technology of Pseudomonas aeruginosa and rhamnolipid, which is applied in the directions of bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of easy contamination of miscellaneous bacteria, difficult to control fermentation process, loss of medium nutrients and bacterial cells, etc. , to achieve the effect of high proportion of disglycolipids, reducing the probability of contamination, and enriching the variety of substrates
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Embodiment 1
[0027] This example illustrates the method for obtaining the strain Pseudomonas aeruginosa YM4 of the present invention.
[0028] The original strain of the bacterium was obtained from the soil around Shengli Oilfield and inoculated into LB liquid medium, and cultivated at 37°C and 200rpm for 12h to obtain a seed solution. Use a pipette gun to take 100 μL of the seed liquid in a sterile test tube of No. 1 900 μL sterile water in the ultra-clean workbench, shake it fully, and disperse the bacterial liquid evenly, that is, 10 -1 sample diluent; take another 10 -1 100 μL of sample diluent in No. 2 sterile test tube of 900 μL sterile water, and so on, mark 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 and 10 -8 . Take 10 respectively -7 and 10 -8 Dilution 200 μL of the dilution solution is evenly coated on the plate with a sterile coating rod, and it is placed flat for 10 minutes, and then it is inverted and cultured in a 37°C constant temperature incubator for 18-24 hours to...
Embodiment 2
[0034] This example illustrates the method for Pseudomonas aeruginosa YM4 provided by the present invention to use glycerol as a carbon source to produce high biosurfactant rhamnolipid.
[0035] Seed medium (LB medium): peptone 10.0g / L, yeast powder 5.0g / L, NaCl 10.0g / L.
[0036] Initial fermentation medium: glycerol 40.0g / L, NaNO 3 2.5g / L, K2 HPO 4 ·3H 2 O 4.0g / L, KH 2 PO 4 4.0g / L, NaCl 1.0g / L, KCl 1.0g / L, MgSO 4 ·7H 2 O 0.2g / L, anhydrous CaCl 2 0.1g / L, yeast powder 1.0g / L, pH 6.5.
[0037] Optimized fermentation medium: glycerol 30.0~60.0g / L, NaNO 3 2.0~12.0g / L,K 2 HPO 4 ·3H 2 O 4.0g / L, KH 2 PO 4 4.0g / L, NaCl 1.0g / L, KCl 1.0g / L, MgSO 4 ·7H 2 O 0.2g / L, anhydrous CaCl 2 0.1g / L, yeast powder 1.0g / L, trace elements 2mL (FeCl 3 0.16g / L, CuSO 4 0.15g / L, ZnSO 4 ·7H 2 O 1.5g / L, MnSO 4 ·H 2 O1.5g / L), pH 7.
[0038] Pseudomonas aeruginosa YM4 provided by the present invention is first activated bacterial classification at 37 ℃ plate, after 20h, a ring of ...
Embodiment 3
[0042] This example illustrates the method for Pseudomonas aeruginosa YM4 to use crude glycerol as a carbon source to high-yield rhamnolipids.
[0043] Seed medium (LB medium): peptone 10.0g / L, yeast powder 5.0g / L, NaCl 10.0g / L.
[0044] Fermentation medium: glycerol 30.0g / L, NaNO 3 12.0g / L,K 2 HPO 4 ·3H 2 O 4.0g / L, KH 2 PO 4 4.0g / L, NaCl 1.0g / L, KCl 1.0g / L, MgSO 4 ·7H 2 O 0.2g / L, anhydrous CaCl 2 0.1g / L, yeast powder 1.0g / L, trace elements 2mL (FeCl 3 0.16g / L, CuSO 4 0.15g / L, ZnSO 4 ·7H 2 O 1.5g / L, MnSO 4 ·H 2 O 1.5g / L), pH 7.
[0045] The content of crude glycerol is 80% (w / w), and 37.5 g of crude glycerol is used per liter of fermentation broth.
[0046] Pseudomonas aeruginosa YM4 provided by the present invention is at first 37 DEG C plate activated bacterial classification, after 20h, connect a ring of well-grown thalli into 50mL LB culture medium, 37 DEG C, 200rpm cultivates 24h, then according to 3% (v / v ) transfer the amount of inoculum into 50mL f...
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